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Circular RNAs (circRNAs) play important roles in carcinogenesis. Here, we investigated the mechanisms and clinical significance of circ-NOL10, a highly repressed circRNA in breast cancer. Subsequently, we also identified RNA-binding proteins (RBPs) that regulate circ-NOL10. Bioinformatics analysis was utilized to predict regulatory RBPs as well as circ-NOL10 downstream microRNAs (miRNAs) and mRNA targets. RNA immunoprecipitation, luciferase assay, fluorescence in situ hybridization, cell proliferation, wound healing, Matrigel invasion, cell apoptosis assays, and a xenograft model were used to investigate the function and mechanisms of circ-NOL10 in vitro and in vivo. The clinical value of circ-NOL10 was evaluated in a large cohort of breast cancer by quantitative real-time PCR. Circ-NOL10 is downregulated in breast cancer and associated with aggressive characteristics and shorter survival time. Upregulation of circ-NOL10 promotes apoptosis, decreases proliferation, and inhibits invasion and migration. Furthermore, circ-NOL10 binds multiple miRNAs to alleviate carcinogenesis by regulating PDCD4. CASC3 and metadherin (MTDH) can bind directly to circ-NOL10 with characterized motifs. Accordingly, ectopic expression or depletion of CASC3 or MTDH leads to circ-NOL10 expression changes, suggesting that these two RBPs modulate circ-NOL10 in cancer cells. circ-NOL10 is a novel biomarker for diagnosis and prognosis in breast cancer. These results highlight the importance of therapeutic targeting of the RBP-noncoding RNA (ncRNA) regulation network., Graphical abstract, A novel RBP-ncRNA signaling route, formed by two RBPs (MTDH and CASC3), one circRNA (circ-NOL10), three miRNAs (miR-149-5p, miR-330-3p, and miR-452-5p), and the effector PDCD4, is identified. This investigation reveals a ncRNA-mediated mechanism in breast cancer carcinogenesis and provides novel insights for therapeutic targeting of ncRNAs.

Metastatic breast cancer is a leading health burden worldwide. Previous studies have shown that metadherin (MTDH) promotes breast cancer initiation, metastasis and therapy resistance; however, the therapeutic potential of targeting MTDH remains largely unexplored. Here, we used genetically modified mice and demonstrate that genetic ablation of Mtdh inhibits breast cancer development through disrupting the interaction with staphylococcal nuclease domain-containing 1 (SND1), which is required to sustain breast cancer progression in established tumors. We performed a small-molecule compound screening to identify a class of specific inhibitors that disrupts the protein–protein interaction (PPI) between MTDH and SND1 and show that our lead candidate compounds C26-A2 and C26-A6 suppressed tumor growth and metastasis and enhanced chemotherapy sensitivity in preclinical models of triple-negative breast cancer (TNBC). Our results demonstrate a significant therapeutic potential in targeting the MTDH–SND1 complex and identify a new class of therapeutic agents for metastatic breast cancer. Kang and colleagues identify a specific compound blocking MTDH1–SND1 interaction, which prevents metastatic breast cancer progression, induces regression of established metastasis in preclinical models and restores chemosensitivity.

Despite increased overall survival rates, curative options for metastatic breast cancer remain limited. We have previously shown that metadherin (MTDH) is frequently overexpressed in poor prognosis breast cancer, where it promotes metastasis and therapy resistance through its interaction with staphylococcal nuclease domain-containing 1 (SND1). Through genetic and pharmacological targeting of the MTDH–SND1 interaction, we reveal a key role for this complex in suppressing antitumor T cell responses in breast cancer. The MTDH–SND1 complex reduces tumor antigen presentation and inhibits T cell infiltration and activation by binding to and destabilizing Tap1/2 messenger RNAs, which encode key components of the antigen-presentation machinery. Following small-molecule compound C26-A6 treatment to disrupt the MTDH–SND1 complex, we showed enhanced immune surveillance and sensitivity to anti-programmed cell death protein 1 therapy in preclinical models of metastatic breast cancer, in support of this combination therapy as a viable approach to increase immune-checkpoint blockade therapy responses in metastatic breast cancer. Kang and colleagues demonstrate that pharmacological targeting of the MTDH1–SND1 axis prevents immune evasion during metastatic progression and provides a synergistic combination strategy with immune-checkpoint blockade to treat metastasis.

Colorectal cancer is the 4th leading cause of cancer related deaths affecting both men and women worldwide. In the present study, any probable role of MTDH mRNA expression in CRC tumorigenesis was explored using both discovery and validation cohorts. After prior ethical and biosafety approvals, tumor tissue samples along with their adjacent controls were collected for this study from Pakistani patients diagnosed with colorectal cancer. RNA was isolated using Trizol reagent, followed by cDNA synthesis. Transcript analysis of MTDH was performed by using qPCR. Moreover, genome-wide expression of MTDH was also determined through micro-array data analysis using BRB-array tools software. MTDH expression was significantly high in tumor tissue samples (p

Prostate cancer (PCa) is one of the most diagnosed cancers in men worldwide. Several studies have identified that circular RNAs (circRNAs) have a crucial impact on the biological processes in PCa. Therefore, it is necessary to study the molecular mechanism of circRNAs in tumor progression and metastasis. RNA interference was used to decrease circHIPK3 and MTDH expression. Overexpression vector was used to increase circHIPK3 and MTDH expression. Luciferase reporter assay were used to detect the relationship between circHIPK3 and miR-448 or between miR-448 and MTDH. MTT assay, colony formation assay and transwell assay were used to measure proliferation and migration of PCa cells. Circular RNA circHIPK3 was significantly increased in PCa tissues and cell lines. And overexpression of circHIPK3 promoted the migration, proliferation, and invasion of PC-3 and 22Rv1 cells, while knockdown of circHIPK3 markedly repressed the above-mentioned series of biological processes. Furthermore, circHIPK3 promoted metadherin (MTDH) expression by sponging miR‐448. In vivo experiments, it was also found that overexpression of circHIPK3 significantly promoted tumor growth. Our research shows that circHIPK3 plays a carcinogenic effect in PCa by regulating the miR-448/MTDH axis, indicating that circHIPK3 may be a potential therapeutic target for PCa.

Background: CircRNA plays an important role in cancer progression. However, the potential mechanism of circRNA in gastric cancer remains unknown. In this study, we aimed to investigate the specific mechanism of circALPL in gastric cancer. Methods: Using a high-throughput microarray, we found that circALPL was upregulated in gastric cancer cell lines. RT-qPCR was used to measure the circALPL expression level in gastric cell lines and tissue. Transwell, CCK-8, and metastasis assays were performed to learn the function after circALPL was inhibited. Results: circALPL downregulation suppresses the invasion and proliferation ability of gastric cancer cells. Additionally, the underlying pathway of circALPL was studied using luciferase reporter assays and RNA immunoprecipitation assays. The results showed that circALPL promotes gastric cancer progression by sponging miR-127, thus upregulating MTDH. Conclusion: The circALPL-miR-127-MTDH pathway plays a vital role in gastric cancer proliferation and metastasis. circALPL might be a new therapeutic target in gastric cancer.

Colorectal and lung cancers account for one-third of all cancer-related deaths worldwide. Previous studies suggested that metadherin (MTDH) is involved in the development of colorectal and lung cancers. However, how MTDH regulates the pathogenesis of these cancers remains largely unknown. Using genetically modified mouse models of spontaneous colorectal and lung cancers, we found that MTDH promotes cancer progression by facilitating Wnt activation and by inducing cytotoxic T-cell exhaustion, respectively. Moreover, we developed locked nucleic acid-modified (LNA) MTDH antisense oligonucleotides (ASO) that effectively and specifically suppress MTDH expression in vitro and in vivo. Treatments with MTDH ASOs in mouse models significantly attenuated progression and metastasis of colorectal, lung, and breast cancers. Our study opens a new avenue for developing therapies against colorectal and lung cancers by targeting MTDH using LNA-modified ASO. Significance: This study provides new insights into the mechanism of MTDH in promoting colorectal and lung cancers, as well as genetic and pharmacologic evidence supporting the development of MTDH-targeting therapeutics.

Xueyan Shi,1 Zhenghao Deng,2 Shouman Wang,3 Shuai Zhao,1,4 Lan Xiao,5 Jiang Zou,1 Tao Li,1 Sichuang Tan,6 SipAin Tan,1 Xianzhong Xiao1 1Sepsis Translational Medicine, Key Laboratory of Hunan Province, Xiangya School of Medicine, Central South University, Changsha, Hunan, 410008, People’s Republic of China; 2Department of Pathology, Xiangya School of Medicine, Central South University, Changsha, Hunan, 410008, People’s Republic of China; 3Department of Breast Surgery, Xiangya Hospital, Central South University, Changsha, Hunan, 410008, People’s Republic of China; 4Department of Pathology, Tianjin Medical University Cancer Institute and Hospital, Tianjin, 300060, People’s Republic of China; 5Department of Traditional Chinese Medicine, The Third Xiangya Hospital, Central South University, Changsha, Hunan, 410008, People’s Republic of China; 6Department of Thoracic Surgery, Second Xiangya Hospital, Central South University, Changsha, Hunan, 410008, People’s Republic of ChinaCorrespondence: SipAin Tan Tel +86 – 13975805079Email springtan@csu.edu.cnSichuang Tan Tel +86 – 13975806723Email strongtan@csu.edu.cnPurpose: To explore the molecular mechanism of promoting cervical cancer by HSF1 in vivo and in vitro.Methods: The expression of HSF1 in 110 paraffin-embedded cervical cancer sections of different grades was examined via immunohistochemistry analyses. Expression of HSF1 downstream targets Metadherin (MTDH), VEGF-C and CD31 were studied using immunohistochemistry analyses. HSF1 transcriptional activity in the MTDH promoter region was detected by EMSA, CHIP and luciferase. Cell proliferation and clonality were detected by MTT and clonal formation assay. Cell migration and invasion ability were investigated by scratch analysis and transwell assay. HSF1-mediated tumorigenesis in vivo was examined in xenograft models.Results: HSF1 expression of cervical cancer cell line was increased compared to normal human cervical tissues. HSF1 enhanced the expression of MTDH, VEGF-C and CD31. HSF1 can combine with MTDH promoter to promote the expression of MTDH. HSF1 enhanced HeLa cell proliferation and clone formation. Furthermore, HSF1 increased HeLa cells migration and invasion in vitro. In the transplanted tumor model, HSF1 inhibited tumor growth in vivo after interference, and reduced the expression of MTDH, VEGF-C and CD31.Discussion: HSF1 can promote the proliferation, metastasis and invasion of cervical cancer.Keywords: HSF1, cervical cancer tissue, metastasis, MTDH, transcriptional regulation

MicroRNAs are a class of small non-coding RNAs that perform a crucial function in posttranscriptional gene regulation. Dysregulation of these microRNAs is associated with many types of cancer progression. In tumorigenesis, downregulated microRNAs might function as a tumour suppressor by repressing oncogenes, whereas overexpressed miRs might function as oncogenes by suppressing tumour suppressor. Similarly, Metadherin (also known as AEG-1/ LYRIC), is an oncogene, the levels of which are found to be very high in various cancers and play a crucial role in the proliferation of cells and invasion. Our review focuses on the study, which shows the alteration of microRNA expression profile and suppression of carcinogenesis when MTDH/AEG-1 is targeted. It summarises the studies where downregulation and upregulation of AEG-1 and microRNAs, respectively, alter the biological functions of the cell, such as proliferation and apoptosis. Studies have reported that AEG-1 can be direct or indirect target of microRNA, which could provide a new-insight to know the underlying molecular mechanism and might contribute to the progress of new therapeutic strategies for the disease.

Staphylococcal nuclease domain-containing protein 1 (SND1) is a multifunctional oncoprotein overexpressed in breast cancer. Binding of metadherin (MTDH) to SND1 results in the stabilization of SND1 and is important in the initiation and progression of breast cancer. Disruption of such interaction is a potential therapeutic for breast cancer. SN1/2 domain of SND1 was used as bait in a phage display screening to identify a 12-amino acid peptide 4-2. The activity of peptide 4-2 was evaluated by ELISA, coimmunoprecipitation, MTS, Western blot analysis, and xenograft mouse model. Peptide 4-2 could disrupt SND1–MTDH interaction. Cell penetrating derivative of peptide 4–2 (CPP-4–2) could penetrate and kill breast cancer cells by disrupting SND1–MTDH interaction and degrading SND1. Tryptophan 10 (W10) of peptide 4-2 was essential in mediating cytotoxicity, SND1 interaction, SND1–MTDH disruption, and SND1 degradation. CPP-4-2 could inhibit the growth of breast cancer in a xenograft mouse model. The SND1-interacting peptide 4-2 could kill breast cancer cells both in vitro and in vivo by interacting with SND1, disrupting SND1–MTDH interaction, and inducing SND1 degradation. W10 was an essential amino acid in the activity of peptide 4-2.

Background & Aims Identifying novel and actionable targets in hepatocellular carcinoma (HCC) remains an unmet medical need. TAK1 was originally identified as a transforming growth factor-β–activated kinase and was further proved to phosphorylate and activate numerous downstream targets and promote cancer progression. However, the role of TAK1 in developed HCC progression and targeted therapy resistance is poorly understood. Methods The expression of TAK1 or MTDH in HCC cell lines, tumor tissues, and sorafenib-resistant models was analyzed by in silico analysis, quantitative real-time polymerase chain reaction, Western blotting, and immunohistochemistry. In vivo and in vitro experiments were introduced to examine the function of TAK1 or MTDH in HCC and sorafenib resistance using small interfering RNA and pharmacologic inhibitors in combination with or without sorafenib. Co-immunoprecipitation and RNA immunoprecipitation were carried out to determine the binding between TAK1 and FBXW2 or between MTDH and FBXW2 mRNA. Protein half-life and in vitro ubiquitination experiment was performed to validate whether FBXW2 regulates TAK1 degradation. Results Our findings unraveled the clinical significance of TAK1 in promoting HCC and sorafenib resistance. We identified a novel E3 ubiquitin ligase, FBXW2, targeting TAK1 for K48-linked polyubiquitylation and subsequent degradation. We also found that MTDH contributes to TAK1 up-regulation in HCC and sorafenib resistance through binding to FBXW2 mRNA and accelerates its degradation. Moreover, combination of TAK1 inhibitor and sorafenib suppressed the growth of sorafenib-resistant HCCLM3 xenograft in mouse models. Conclusions These results revealed novel mechanism underlying TAK1 protein degradation and highlighted the therapeutic value of targeting TAK1 in suppressing HCC and overcoming sorafenib resistance.

Protein-metabolite interactions (PMIs) play important roles in various biological processes, especially in disease progression. However, due to the complexity of living cells, it is very difficult to identify specific PMIs. Herein, we chose one oncogenic factor, metadherin (MTDH), as a bait to identify its in vivo interacting metabolites in cancer cells. Cholesterol is an important metabolite and essential structural component of cell membranes. It could also drive several diseases including cancer. Interestingly, we found that cholesterol robustly interacted with MTDH and downregulated the expression of MTDH in cancer cells. Furthermore, MTDH disturbed metabolite alterations under cholesterol treatment in MTDH transduced cancer cells. Collectively, our results uncover an undescribed PMI where MTDH, as an oncogenic factor, might positively regulate cancer progression by interacting with cholesterol. This study interprets the theoretical basis of PMI-oriented cancer progression and targeting therapies in clinic.

Long non-coding RNAs (lncRNAs) have recently emerged as key regulators of the occurrence and progression of various human cancers, including colorectal cancer. However, the regulatory mechanism of lncRNAs in the tumorigenesis of colorectal cancer remains poorly understood. In this study, we aimed to elucidate the potential role of lncRNA HCG18 in colorectal cancer. Herein, we found that HCG18 expression was significantly upregulated in colorectal cancer tissues and cell lines. Knockdown of HCG18 significantly inhibited the growth and invasion of colorectal cancer cells, while its overexpression had the opposite effect. Moreover, HCG18 was identified as a sponge of miR-1271. Our results showed that knockdown of HCG18 markedly upregulated miR-1271 expression in colorectal cancer cells. Notably, HCG18 expression was inversely correlated with miR-1271 expression in colorectal cancer specimens. Further investigation revealed that HCG18 contributed to the enhancement of MTDH/Wnt/β-catenin signalling in colorectal cancer cells. The antitumour effect of HCG18 inhibition was significantly reversed by miR-1271 inhibition or MTDH overexpression. Overall, the results of our study demonstrate that HCG18 exerts a potential oncogenic function in colorectal cancer by enhancing MTDH/Wnt/β-catenin signalling via sponging of miR-1271, highlighting the importance of HCG18/miR-1271/ MTDH/Wnt/β-catenin signalling in the progression of colorectal cancer.

Neuroblastoma (NB) is responsible for 15% of all childhood cancer deaths. Despite advances in treatment and disease management, the overall 5-year survival rates remain poor in high-risk disease (25-40%). It is well known that miR-145 functions as a tumor suppressor in several types of cancer. However, the impact of miR-145 on NB is still ambiguous. Our aim was to investigate the potential tumor suppressive role and mechanisms of miR-145 in high-risk neuroblastoma. Expression levels of miR-145 in tissues and cells were determined using RT-qPCR. The effect of miR-145 on cell viability was evaluated using MTT assays, apoptosis levels were determined using TUNEL staining, and the MTDH protein expression was determined using western blot and RT-PCR. Luciferase reporter plasmids were constructed to confirm direct targeting for MTDH. The results showed that miR-145 expression was significantly lower in high-risk MYCN amplified (MNA) tumors and low miR-145 expression was associated with worse EFS and OS in our cohort. Over-expression of miR-145 reduced cell viability and increased apoptosis in SH-SY-5Y cells. We identified MTDH as a direct target for miR-145 in SH-SY-5Y cells. Targeting MTDH has the similar results as miR-145 overexpression. Our findings suggest that low miR-145 expression was associated with poor prognosis in patients with NB, and the overexpression of miR-145 inhibited NB cells growth by down-regulating MTDH, thus providing a potential target for the development of microRNA-based approach for NB therapy.

Cell-free circular RNAs (circRNAs) stably and abundantly exist in body fluids. In this study we aimed to investigate the potential of urinary cell-free circRNAs as a novel class of noninvasive disease biomarkers for diagnosis of bladder cancer. Differentially expressed circRNAs from 10 normal and 10 bladder cancer urine samples were firstly detected by microarray. Hsa_circ_0137439 was then screened and validated in 30 normal and 116 bladder cancer samples. A receiver operating characteristic (ROC) curve was used to evaluate the diagnostic value of hsa_circ_0137439. The Kaplan-Meier method was used to evaluate the significance of hsa_circ_0137439 in the prognosis of bladder cancer. We found that hsa_circ_0137439 was significantly upregulated in bladder cancer samples. Moreover, increased expression of hsa_circ_0137439 was correlated with higher tumor stage, higher tumor grade, higher lymph node status, and history of muscle-invasive bladder cancer (MIBC). Also, urinary cell-free hsa_circ_0137439 could not only differentiate bladder cancer from normal controls but also distinguish MIBC from non-muscle-invasive bladder cancer (NMIBC). Additionally, hsa_circ_0137439 in urine supernatant could serve as an independent prognostic predicator of recurrence-free survival and overall survival for patients with bladder cancer. Cell assays showed that hsa_circ_0137439 knockdown contributed to the inhibition of cell proliferation and migration via hsa_circ_0137439/miR-142-5p/ MTDH axis. In conclusion, urinary cell-free hsa_circ_0137439 could be a promising biomarker for tumor diagnosis and prognostic assessment of bladder cancer patients.

Metadherin (MTDH) has been recognized as a novel protein that is critical for the progression of multiple types of human malignancies. Studies have reported that MTDH enhances the metastatic potential of cancer cells by regulating multiple signaling pathways. miRNAs and various tumor-related proteins have been shown to interact with MTDH, making it a potential therapeutic target as well as a biomarker in human malignancies. MTDH plays a critical role in inflammation, angiogenesis, hypoxia, epithelial–mesenchymal transition and autophagy. In this review, we present the function and mechanisms of MTDH for cancer initiation and progression.

Summary Immune checkpoint blockade (ICB) persistently provides a prognosis improvement but only in a small fraction of patients with cancer due to immunotherapy resistance induced by the consecutive activated oncogenic pathways, including MAPK, Akt, and WNT pathway partially driven by Metadherin (MTDH). However, there is no study to investigate the potential role and mechanisms of MTDH in ICB-treated cancers. Here, we systematically explored the cohorts from The Cancer Genome Atlas (TCGA) and independent cancer cohorts. Elevated MTDH expression was founded to associate with a worse overall survival and poorer immune response in patients with cancer. Dysregulated tumor-infiltrating immune cells and inhibitory immune checkpoint expression were correlated with MTDH expression. Furthermore, the mutual interactions between epithelial-to-mesenchymal-transition, m6A-RNA-methylation, and MTDH may illustrate the potential mechanisms of MTDH resistant to ICB treatment. Although more designed experiments and trials are needed in the future, targeting MTDH may help to overcome immunotherapy resistance in a wide range of cancers., Graphical abstract, Highlights • MTDH associates with prognosis and immunotherapy response for patients with cancer • MTDH associates with dysregulated tumor immune environment and checkpoint expression • The MTDH/m6A RNA methylation/EMT pathway may contribute to immunotherapy resistance, Biological sciences; Molecular biology; Expression study

Medulloblastoma is a pediatric brain tumor arising from the cerebellum and brainstem that is curable with surgery and aggressive cytotoxic therapy including craniospinal irradiation and chemotherapy. Preclinical models indicate that a small pool of de-differentiated, stem cell-like medulloblastoma cells are resistant to cytotoxic treatment and contribute to medulloblastoma relapse after aggressive therapy. Here, we identify Miat as a Shh and Myc regulated long noncoding RNA (lncRNA) that is required for maintenance of a treatment-resistant medulloblastoma stem-like phenotype. Loss of Miat delays medulloblastoma formation in genetically defined mouse models of Shh medulloblastoma and enforces differentiation of tumorigenic stem-like medulloblastoma cells into a non-tumorigenic cell state. Miat facilitates treatment resistance in part by downregulating p53 signaling and impairing radiation induced cell death in stem-like MB cells, which can be reversed by therapeutic inhibition of Miat with antisense oligonucleotides. The RNA binding protein Metadhedrin (Mtdh), which is associated with resistance to cytotoxic therapy in numerous types of cancer, co-localizes with Miat in stem-like medulloblastoma cells. Further, loss of Mtdh activates p53 signaling and reduces tumorigenicity in stem-like medulloblastoma cells. Taken together, these data reveal a critical role for the lncRNA Miat in sustaining a treatment resistant pool of tumorigenic stem-like medulloblastoma cells.

This study explored the role of MTDH in regulating the sensitivity of breast cancer cell lines to gemcitabine (Gem) and the potential miRNAs targeting MTDH. The expression of MTDH in cancer tissues and cells was detected by immunohistochemical staining or qRT-PCR. The target genes for MTDH were predicted by bioinformatics and further confirmed by dual-luciferase reporter assay and qRT-PCR. Cancer cells were transfected with siMTDH, MTDH, miR-9-3p inhibitor, or mimics and treated by Gem, then CCK-8, colony formation assay, tube formation assay, flow cytometry, wound healing assay, and Transwell were performed to explore the effects of MTDH, miR-9-3p, and Gem on cancer cell growth, apoptosis, migration, and invasion. Expressions of VEGF, p53, cleaved caspase-3, MMP-2, MMP-9, E-Cadherin, N-Cadherin, and Vimentin were determined by Western blot. MTDH was high-expressed in cancer tissues and cells, and the cells with high-expressed MTDH were less sensitive to Gem, while silencing MTDH expression significantly promoted the effect of Gem on inducing apoptosis, inhibiting cell migration, invasion, and growth, and on regulating protein expressions of cancer cells. Moreover, miR-9-3p had a targeted binding relationship with MTDH, and overexpressed miR-9-3p greatly promoted the toxic effects of Gem on cancer cells and expressions of apoptosis-related proteins, whereas overexpressed MTDH partially reversed such effects of overexpressed miR-9-3p. The study proved that miR-9-3p regulates biological functions, drug resistance, and the growth of Gem-treated breast cancer cells through targeting MTDH.

Intestinal barrier dysfunction is characterized by increased intestinal permeability to lumen endotoxin, showing remarkable predisposition to immune enteropathy, and colorectal cancer tumor necrosis factor (TNF)-α is associated with this pathological process, while the mechanism remains unknown. In this study, different doses of TNF-α were used for Caco-2 cell treatment. We discovered that miR-21-3p expression was obviously increased by TNF-α in a dose-dependent manner. Further study demonstrated that TNF-α could upregulate miR-21-3p expression through the NF-κB signaling pathway. Then, TargetScan and miRWalk miRNA–mRNA interaction prediction online tools were introduced, and metadherin (MTDH) was screened out as a potential target of miR-21-3p. We subsequently found that miR-21-3p could directly target the 3′-untranslated region (UTR) of MTDH mRNA and inhibit its expression. Furthermore, it was demonstrated that miR-21-3p could regulate the Wnt signaling pathway by targeting MTDH mRNA, suggesting the effect of miR-21-3p/MTDH/Wnt axis on intestinal barrier dysfunction. Our findings provide a novel potential biomarker and therapeutic target for intestinal barrier dysfunction and related diseases.

In recent years, there have been reports about the involvement of circular RNAs (circRNAs) in the pathogenesis of gastric cancer (GC), but the molecular mechanism in cell proliferation, invasion, and migration is still unclear. Based on The Cancer Genome Atlas (TCGA) database, we analyzed differentially expressed circRNAs between GC and non-tumor tissues. Gene Ontology and Kyoto Encyclopedia of Genes and Genomes enrichment analysis were used to clarify the functional role in GC. Here, we showed that circITGA7 was lowly expressed in GC tissues based on the TCGA database. In vitro, silencing the expression of circITGA7 increased cell proliferation and metastasis, whereas overexpression did the opposite. Mechanistically, miR-1471 has circITGA7 as a sponge, and miR-1471 has metadherin (MTDH) as a target gene. Consequently, functional analysis showed that the tumor suppressor effect of circITGA7 was the result of regulating the miR-1471/MTDH axis. Overall, the circITGA7/miR-1471/MTDH signaling pathway may play a crucial role in GC, providing a new potential mechanism involved in GC progression.

BACKGROUND Breast cancer is a common malignant tumor worldwide. Despite the huge advances in modern medicine, many patients still face a high risk of recrudescent and metastatic breast cancer. Berberine was widely implemented in clinic treatment of breast cancer. This study was performed to contribute to a better understanding on the mechanisms underlying berberine affecting breast cancer. MATERIAL AND METHODS We mined survival data of metadherin (MTDH) in breast cancer patients through Kaplan-Meier Plotter and analyzed the transcriptional and posttranscriptional expression profile of MTDH in several breast cancer cell lines. The cell viability and MTDH mRNA level were detected under the si-MTDH vector and different concentrations of berberine. The MTDH-expression vector was transfected into MCF-7 and MDA-MB-231 cells, and the changes of cell viability and apoptosis were determined after berberine (50 μM) treatment. RESULTS High MTDH expression was related to worse relapse-free survival (RFS) of breast cancer (P-value=6.2e-08). High-expressed MTDH is common in breast cancer cells, compared with that in normal breast cells (P

Cisplatin (CDDP)-based chemotherapy is the gold standard treatment for many types of cancer. However, the phenotypic hallmark of tumors often changes after CDDP treatment, with the acquisition of epithelial-to-mesenchymal transition (EMT) and platinum resistance. Furthermore, the mechanisms by which cancer cells acquire EMT under the control of CDDP remain unclear. Following an investigation of urothelial carcinoma (UC) before and after the acquisition of platinum resistance, we offer the new target TNFAIP2, which led to EMT and tumor invasion in platinum-treated UC cells. TNFAIP2 expression in cancer was examined at the protein and transcriptional levels. A potential target for TNFAIP2 during EMT was assessed by microarray. Clinically, upregulated TNFAIP2 expression was identified as a significant predictor of mortality following surgery in three different cohorts of patients with UC (n = 156, n = 119, and n = 54). Knockdown of TNFAIP2 resulted in upregulation of E-cadherin expression and downregulation of TWIST1 expression, which decreased motile function in platinum-resistant UC cells. TNFAIP2 overexpression led to downregulation of E-cadherin expression and upregulation of TWIST1 expression in platinum-naïve UC cells. Clinical investigation of matched pre- and post-CDDP-treated UC sections confirmed upregulation of TNFAIP2 expression in CDDP-treated tumors but downregulation of E-cadherin expression. Global gene expression analysis following TNFAIP2 knockdown identified MTDH as a positive regulator of TNFAIP2-derived EMT acquisition in cancer cells. The present results suggest a relationship between TNFAIP2 and EMT in cancers under the control of CDDP, in which MTDH expression levels in cancer cells are vital for promoting TNFAIP2-derived EMT acquisition.

Purpose Metadherin (MTDH), as an oncogene, is associated with metastasis and poor prognosis. This study investigated MTDH expressions and development of gastric cancer (GC) cell phenotypes and the contribution of MTDH to epithelial-mesenchymal transition (EMT). Patients and methods MTDH expression was assayed in human GC cell lines and tumor tissue from 92 GC patients. Functional experiments were performed to characterize MTDH activity. Expressions of EMT-related proteins (vimentin and E-cadherin), phosphorylated β-catenin and β-catenin were assayed by immunohistochemistry, Western blotting, immunofluorescence, and co-immunoprecipitation, respectively. Results MTDH expressions were higher in GC tissue than that in gastric mucosa from the same patient. MTDH overexpression was correlated with metastasis and enhanced malignant GC phenotypes, i.e., proliferation, migration, invasiveness, and chemoresistance. MTDH overexpression was associated with expressions of vimentin, E-cadherin and cancer stem-cell biomarkers including CD44, CD133, and Oct4. MTDH complexed with β-catenin and decreased phosphorylated β-catenin levels to facilitate β-catenin translocation into the nucleus and expressions of downstream genes. Conclusion MTDH overexpression in GC cells is associated with EMT and development of cancer stem cell malignant phenotypes and affects the subcellular translocation of β-catenin. The results warrant investigation of the prognostic value of MTDH in GC and as a therapeutic target.

Background: MicroRNA-876-5p (miR-876) dysregulation contributes to the aggressiveness of various types of human cancer. This study was aimed at measuring miR-876 expression in breast cancer, determining the specific roles of miR-876 in the progression of breast cancer and understanding the corresponding molecular mechanisms. Materials and methods: miR-876 expression in breast cancer tissues and cell lines was quantified via RT-qPCR. The effect of miR-876 upregulation on the malignant phenotype of breast cancer cells was investigated using CCK-8 assays, flow cytometry, Transwell migration and invasion assays and tumor xenograft experiments. The mechanisms underlying the tumor-suppressive action of miR-876 in breast cancer cells were explored using bioinformatic analysis, luciferase reporter assays, RT-qPCR and Western blot analysis. Results: miR-876 was found to be underexpressed in breast cancer tissues and cell lines. Decreased miR-876 expression notably correlated with lymphatic invasion metastasis, TNM stage and differentiation grade. Overall survival was lower among patients with breast cancer and low miR-876 expression than in patients with high miR-876 expression. Restoration of miR-876 expression decreased breast cancer cell proliferation, migration and invasion in vitro and restricted tumor growth in vivo as well as increased cell apoptosis. Metadherin (MTDH) was identified as a novel target of miR-876 in breast cancer cells. Furthermore, long intergenic nonprotein-coding RNA 707 (LINC00707) acted as a molecular sponge for miR-876, thereby regulating MTDH expression in breast cancer. Finally, silencing miR-876 expression attenuated the influence of a LINC00707 knockdown on the malignancy of breast cancer cells. Conclusion: This study, thus, revealed the vital functions of the LINC00707-miR-876-MTDH pathway in breast cancer and provided attractive targets and markers for its treatment.

Studies have rarely been conducted on the role of miRNAs in prostate cancer (PCa) cell progression by directly targeting MTDH, at least to the best of our knowledge. Thus, the present study aimed to identify miRNAs closely related with metadherin (MTDH) and to determine their roles in PCa. For this purpose, the expression levels of MTDH in PCa tissues and cell lines were examined by RT‑qPCR, immunohistochemistry and western blot analysis. By cell transfection, MTDH was either overexpressed in the normal prostate epithelial cell lines or silenced in tumor cell lines to determine cell viability, invasion and migration. Bioinformatics analysis, RT‑qPCR, western blot analysis, dual‑luciferase reporter assay and MTT assay were performed to identify direct the target of MTDH and to examine tumor cell viability. Rescue experiments using the PC‑3 and LNCaP cells were carried out by MTT assay, scratch wound assay, Transwell assay, RT‑qPCR and western blot analysis. Experiments were also conducted using 46 PCa human cancer and adjacent tissues, as wells as on 501 cases of PCa from the TCGA database. It was confirmed that the overexpression of MTDH was associated with a poor prognosis of patients. The overexpression of MTDH was found to promote the viability, invasion and migration of PCa cells. miR‑145‑5p and miR‑145‑3p identified from 16 miRNAs were found to be closely related to PCa and to be the targets of MTDH. Both these miRNAs were found to significantly suppress the growth and metastasis of PCa cells by negatively regulating the expression of MTDH. On the whole, the findings of this study demonstrate that MTDH functions as an oncogene in PCa and the inhibition of MTDH by miR‑145‑5p or miR‑145‑3p suppressed the growth and metastasis of PCa cells. The miR‑145‑5p/MTDH and miR‑145‑3p/MTDH pathways may thus become novel treatment targets for PCa.

Metastasis of ovarian cancer is regulated by microRNAs. This study focused on the effects of miR-30a-5p on ovarian cancer migration and invasion. Our results showed that the miR-30a-5p and mucin type O-glycan biosynthesis are closely related to ovarian cancer, and that miR-30a-5p was downregulated in ovarian cancer cells. miR-30a-5p overexpression reduced cell viability and inhibited migration and invasion in HO-8910 and HO-8910PM cells. S phase kinase-associated protein 2 (SKP2), B cell lymphoma 9 (BCL9), and NOTHC1 are direct target genes of miR-30a-5p. MTDH, SKP2, BCL9, and NOTCH1 genes were overexpressed in ovarian cancer cells, and they are direct target genes of miR-30a-5p. miR-30a-5p overexpression inhibited epithelial-mesenchymal transition (EMT) process, while upregulation of SKP2, BCL9, and NOTCH1 gene expression levels reduced the inhibition of EMT process by miR-30a-5p. miR-30a-5p was lowly expressed in ovarian cancer, and such a phenomenon is related to ovarian cancer metastasis. miR-30a-5p might inhibit the migration and invasion of ovarian cancer cells by downregulating the expression of SKP2, BCL9, and NOTCH1 genes.

Metastasis is one of the primary causes for high mortality in patients with squamous cell carcinoma of the head and neck (SCCHN). Our previous study showed that chemokine (C‐C motif) ligand 18 (CCL18), derived from tumour‐associated macrophages (TAMs), regulates SCCHN metastasis by promoting epithelial‐mesenchymal transition (EMT) and preserving stemness. However, the underlying mechanism needs to be further investigation. Interestingly, metadherin (MTDH) expression was induced when SCCHN cells were stimulated with recombinant CCL18 protein in this study. Suppressing MTDH expression reversed CCL18‐induced migration, invasion and EMT in SCCHN cells. Furthermore, the NF‐κB signalling pathway was involved in the MTDH knock‐down cells with CCL18 stimulation. We performed ELISA to evaluate the CCL18 levels in the serums of 132 treatment‐naive SCCHN patients, 25 patients with precancerous lesion and 32 healthy donors. Our results demonstrated that serum CCL18 levels were significantly higher in SCCHN patients than patients with precancerous lesion and healthy individuals. CCL18 levels were found to be significantly correlated with tumour classification, clinical stage, lymph node metastasis and histological grade in SCCHN patients. Thus, our findings suggest that CCL18 may serve as a potential biomarker for diagnosis of SCCHN and promote SCCHN invasion, migration and EMT by MTDH‐NF‐κB signalling pathway.

Macrophages in the intestinal mucosa can rapidly engage Toll-like receptor [TLR]-mediated inflammatory responses to protect against pathogen invasion, but these same innate immune responses can also drive the induction of colitis. Our previous research revealed that metadherin [MTDH] is overexpressed in multiple cancers and plays vital roles in tumour progression. However, the role of MTDH in intestinal inflammation is largely unknown. In this study, we found the MTDH expression in colonic lamina propria [CLP] macrophages was positively correlated with inflammatory colitis severity. MTDH-/- mice were protected against the symptoms of dextran sodium sulphate [DSS]-induced colitis; however, adoptive transfer of MTDH wild-type [WT] monocytes partially restored the susceptibility of MTDH-/- mice to DSS-induced colitis. TLR stimulation was sufficient to induce the expression of MTDH, whereas the absence of MTDH was sufficient to suppress TLR-induced production of inflammatory cytokines by macrophages. From a mechanistic perspective, MTDH recruited TRAF6 to TAK1, leading to TRAF6-mediated TAK1 K63 ubiquitination and phosphorylation, ultimately facilitating TLR-induced NF-κB and MAPK signalling. Taken together, our results indicate that MTDH contributes to colitis development by promoting TLR-induced pro-inflammatory cytokine production in CLP macrophages and might represent a potential therapeutic approach for intestine inflammation intervention.

BACKGROUND Head and neck cancer (HNSCC) is one of the most lethal cancers characterized by high relapse and poor prognosis. Several miRNAs have been implicated in HNSCC, including the tumor suppressor miR-137. A large CpG island (CpG73) spans most of the miR-137 gene sequence and stretches 659-bp downstream, ending just upstream of miR-2682 in the same host gene. Here, we assessed the role of the MIR137/MIR2682 locus in HNSCC. METHODS MiRNA expression was analyzed in paired cancerous and normal tissues from 77 HNSCC patients by Quantitative Reverse-Transcription PCR. CpG73 methylation in paired tissues from 48 patients was determined by combined bisulfite restriction analysis. Associations between expression and methylation levels and patient clinicopathological parameters were investigated. RESULTS Decreased expression of miR-137 (P

Since its initial cloning in 2002, a plethora of studies in a vast number of cancer indications, has strongly established AEG-1 as a bona fide oncogene. In all types of cancer cells, overexpression and knockdown studies have demonstrated that AEG-1 performs a seminal role in regulating proliferation, invasion, angiogenesis, metastasis and chemoresistance, the defining cancer hallmarks, by a variety of mechanisms, including protein-protein interactions activating diverse oncogenic pathways, RNA-binding promoting translation and regulation of inflammation, lipid metabolism and tumor microenvironment. These findings have been strongly buttressed by demonstration of increased tumorigenesis in tissue-specific AEG-1 transgenic mouse models, and profound resistance of multiple types of cancer development and progression in total and conditional AEG-1 knockout mouse models. Additionally, clinicopathologic correlations of AEG-1 expression in a diverse array of cancers establishing AEG-1 as an independent biomarker for highly aggressive, chemoresistance metastatic disease with poor prognosis have provided a solid foundation to the mechanistic and mouse model studies. In this review a comprehensive analysis of the current and up-to-date literature is provided to delineate the clinical significance of AEG-1 in cancer highlighting the commonality of the findings and the discrepancies and discussing the implications of these observations.

Vasculogenic mimicry (VM) refers to vessel-like structures formed by aggressive tumor cells and is closely associated with cancer invasion and metastasis. Here, we investigated the effect of macrophage-derived MTDH on VM formation in head and neck squamous cell carcinoma (HNSCC) and its underlying mechanism. Macrophages with MTDH overexpression (Mac-MTDH) promoted cancer cell VM formation, migration, and invasion in vitro. Moreover, MTDH overexpression triggered macrophage polarization into M2 type tumor-associated macrophages. Analysis of HNSCC clinical samples revealed that MTDH+ macrophages were predominantly located in the tumor-stromal region in proximity to VM and correlated with lymph node metastasis. Mechanistically, Mac-MTDH enhanced the expression and secretion of VEGFA-165 rather than other VEGFA isoforms via s-catenin. The VEGFA-165/Flt-1 axis was responsible for Mac-MTDH’s effects in cancer cells through p-STAT3/Twist1/VE-cadherin pathway. Using mouse model, we further confirmed that Mac-MTDH increased VM formation and cancer metastasis in vivo. Furthermore, in subcutaneous xenograft mouse model, HN6 + Mac-MTDH tumor exhibited elevated expression of p-STAT3 and Twist1 than HN6 + Mac-NC tumors. This study revealed that Mac-MTDH promoted VM formation, cancer cell migration and invasion, and cancer metastasis through VEGFA-165/Flt-1 axis, and that macrophage-derived MTDH could be a potential therapeutic target in HNSCC.

BackgroundIdentifying novel and actional targets in hepatocellular carcinoma (HCC) remains an unmet medical need. TAK1 was originally identified as a TGF-β-activated kinase and was further proved to phosphorylate and activate numerous downstream targets and promote cancer progression. Although TAK1 depletion leads to early onset of hepatocarcinogenesis in mice, the role of TAK1 in developed HCC progression and targeted therapy resistance is poorly understood. MethodsThe expression of TAK1 and MTDH in HCC cell lines and patients and sorafenib-resistant models was analyzed by in silico analysis, quantitative real-time PCR, western blotting and immunohistochemistry. In vivo and In vitro experiments was introduced to examine the function of TAK1 or MTDH in HCC and sorafenib resistance using small interfering RNA and pharmacological inhibitors in combination with or without sorafenib. Co-immunoprecipitation and RNA immunoprecipitation was carried out to determine the binding between TAK1 and FBXW2 or between MTDH and FBXW2 RNA.ResultsOur findings unraveled the clinical significance of TAK1 in promoting HCC and sorafenib resistance. We identified a novel E3 ubiquitin ligase, FBXW2, targeting TAK1 for K48-linked polyubiquitylation and subsequent degradation. We also found that MTDH contributes to TAK1 upregulation in HCC and sorafenib resistance, through binding to FBXW2 mRNA and accelerate its degradation. Moreover, combination of TAK1 inhibitor and sorafenib suppressed the growth of sorafenib-resistant HCCLM3 xenograft in mouse models.ConclusionThese results revealed novel mechanism underlying TAK1 protein degradation and highlighted the therapeutic value of targeting TAK1 in suppressing HCC and overcoming sorafenib resistance.

[ES] El cáncer de mama es el cáncer más común en mujeres en todo el mundo y la principal causa de muerte por cáncer en mujeres junto al cáncer de pulmón. Este cáncer tiene muy buen pronóstico en general, con una supervivencia del 80%. Sin embargo, el pronóstico del cáncer de mama triple negativo es mucho peor, al no conocerse ninguna diana farmacológica y tratarse de forma inespecífica. La metformina, fármaco prescrito para la diabetes, ha mostrado algunos buenos resultados preliminares como potencial terapia. Por otro lado, el principal tratamiento dirigido de las pacientes HER2+ es el trastuzumab, que neutraliza al receptor HER2 amplificado; sin embargo, un elevado número de pacientes desarrollan resistencias al tratamiento. Los microRNAs son pequeños RNAs no codificantes capaces de regular la expresión génica epigenéticamente, y pueden ser secretados de la célula en vesículas llamadas exosomas. El objetivo de este trabajo es abordar estas dos problemáticas en cáncer de mama. Son necesarios estudios de los mecanismos de acción o resistencia de estos fármacos a través de la regulación epigenética por microRNAs. Queremos determinar la relación del miR-26a y sus dianas con el efecto de la metformina en cáncer de mama triple negativo y estudiar las diferencias de expresión de microRNAs que generan resistencias a trastuzumab en cáncer de mama HER2+, así como estudiar su modo de transmisión entre células. Se realizaron ensayos celulares tratando con metformina las líneas MDA-MB-231, MDA-MB-468 y MCF-7 así como sobreexpresando o inhibiendo miR-26a y se midieron sus dianas teóricas por qPCR. Para las líneas HER2+ se realizó un Affymetrix Genechip miRNA 4.0 microarray comparando líneas SKBR-3wt y BT-474wt con sus respectivas líneas con resistencia generada a trastuzumab y HCC-1954 como resistente innata. Se estudiaron los microRNAs más relevantes del array en las líneas celulares y en pacientes y se comprobó su presencia en exosomas, así como el efecto de los exosomas en la transmisión de la resistencia. La sobreexpresión de miR-26a resultó en una reducción en la viabilidad celular que se recuperó parcialmente al inhibirla. E2F3, MCL-1, EZH2, MTDH y PTEN fueron regulados negativamente por miR-26a y la proteína PTEN también se redujo tras la sobreexpresión de miR-26a. El tratamiento con metformina redujo la viabilidad de las células de cáncer de mama, aumentó la expresión de miR-26a y condujo a una reducción en la expresión de BCL-2, EZH2 y PTEN. La inhibición de miR-26a previene parte del efecto en viabilidad de la metformina y la reducción de la expresión de PTEN y EZH2. En las líneas HER2+, miR-23b-3p y miR-146a-5p fueron los principales candidatos extraídos del array. miR-23b-3p inhibió PTEN significativamente en la línea BT-474. miR-146a-5p aumentó la resistencia de las células SKBR-3 al trastuzumab y su inhibición redujo la resistencia de las SKBR-3r. El aumento de miR-146a-5p en SKBR-3wt tuvo un efecto en ciclo celular aumentando la fase S y la G2/M, inhibiendo la expresión de CDKN1A y aumentando la de CCNB1. Los exosomas de las SKBR-3 contenían miR-146a-5p, con mayores niveles en los de las resistentes (exoR). Los exoR aumentaron la resistencia a trastuzumab, la transición epitelio-mesenquimal y la migración al co-cultivarse con SKBR-3wt, y la angiogénesis en las HUVEC. Nuestros resultados sugieren que el efecto de la metformina está mediado por una mayor expresión de miR-26a y reducción de sus dianas, PTEN y EHZ2. Por tanto, el uso de metformina en el tratamiento del cáncer de mama constituye una prometedora potencial terapia. En HER2+, miR-23b parece provocar resistencia a trastuzumab vía PTEN y miR-146a a través del ciclo celular. Además, miR-146a se transmite en exosomas, que son capaces de reducir la sensibilidad al trastuzumab de las células sensibles y aumentar la TEM, migración y angiogénesis., [EN] Breast cancer is the most common cancer in women worldwide and the leading cause of cancer death in women along with lung cancer. This cancer has a very good general prognosis, with a survival of 80%. However, the prognosis for triple negative breast cancer is much worse, as it has no pharmacological target and treats it nonspecifically. Metformin, a prescribed diabetes drug, has shown some good preliminary results as potential therapy. On the other hand, the main targeted treatment for HER2 + patients is trastuzumab, which neutralizes the amplified HER2 receptor, but a large number of patients experienced resistance to treatment. MicroRNAs are small non-coding RNAs that are part of epigenetics and are capable of regulating gene expression, and which can be secreted from the cell into vesicles called exosomes. The objective of this work is to address these two problems in breast cancer, which need to study the mechanism of action or resistance of these drugs, through the epigenetics of microRNAs. We want to determine the relationship of miR-26a and its targets with the effect of metformin in triple negative breast cancer and to study the differences in the expression of microRNAs that process resistance to trastuzumab in HER2 + breast cancer, as well as to study its mode of transmission between cells. Cellular assays were performed treating the MDA-MB-231, MDA-MB-468 and MCF-7 lines with metformin as well as overexpressing or inhibiting miR-26a, and their theoretical targets were measured by qPCR. For the HER2+ cell lines, an Affymetrix Genechip miRNA 4.0 microarray was performed comparing SKBR-3wt and BT-474wt lines with their respective cell lines with generated resistance to trastuzumab and HCC-1954 as innate resistance. The most relevant microRNAs of the array in cell lines and in patients were studied and their presence in exosomes was verified, as well as the effect of exosomes in the transmission of resistance. The overexpression of miR-26a resulted in a reduction in cell viability that was partially recovered by inhibiting it. E2F3, MCL-1, EZH2, MTDH, and PTEN were down-regulated by miR-26a, and the PTEN protein was also reduced after overexpression of miR-26a. Metformin treatment reduced the viability of breast cancer cells, increased miR-26a expression, and led to a reduction in BCL-2, EZH2, and PTEN expression. Inhibition of miR-26a partly prevents the effect of metformin in viability and the reduction of the expression of PTEN and EZH2. In the HER2+ lines, miR-23b-3p and miR-146a-5p were the main candidates extracted from the array. miR-23b-3p was shown to significantly inhibit PTEN in the BT-474 cell line. miR-146a-5p increased resistance of SKBR-3wt cells to trastuzumab and its inhibition reduced resistance of SKBR-3r. The increase of miR-146a-5p in SKBR-3wt had effect on the cell cycle by increasing the S phase and the G2/M, inhibiting the expression of CDKN1A and increasing CCNB1 levels. Exosomes isolated from SKBR-3 cell lines contained miR-146a-5p, with higher levels in exosomes from the resistant cell line (exoR). The exoR were shown to increase trastuzumab resistance, EMT, and migration when co-cultivated with SKBR-3wt, and angiogenesis when in culture with HUVEC. Our results indicate that metformin effectively reduces breast cancer cell viability and suggests that the effects of the drug are mediated by an increase in miR-26a expression and a reduction of its targets, PTEN and EHZ2. Thus, the use of metformin constitutes a promising potential triple negative breast cancer therapy. In HER2+ breast cancer, miR-23b appears to elicit resistance to trastuzumab via PTEN and miR-146a throughout the cell cycle. Furthermore, miR-146a is transmitted in exosomes, which have been shown to reduce the sensitivity to trastuzumab of sensitive cells and increase EMT, migration, and angiogenesis., [CA] El càncer de mama és el càncer més comú en dones arreu del món i la principal causa de mort per càncer en dones junt amb el càncer de pulmó. Aquest càncer té molt bon pronòstic en general, amb una supervivència del 80%. No obstant això, el pronòstic del càncer de mama triple negatiu és molt pitjor, al no conèixer-se'n cap diana farmacològica i tractar-se de forma inespecífica. La metformina, fàrmac prescrit per a la diabetis, ha mostrat alguns bons resultats preliminars com a potencial teràpia. D'altra banda, el principal tractament dirigit de les pacients HER2+ és el trastuzumab, que neutralitza el receptor HER2 amplificat; tanmateix, un elevat nombre de pacients desenvolupen resistències al tractament. Els microRNAs són xicotets RNAs no codificants capaços de regular l'expressió gènica epigenèticament, i poden ser secretats de la cèl·lula en vesícules anomenades exosomes. L'objectiu d'aquest treball és abordar aquestes dues problemàtiques en càncer de mama. Són necessaris estudis dels mecanismes d'acció o resistència d'aquests fàrmacs a través de la regulació epigenètica per microRNAs. Volem determinar la relació del miR-26a i les seues dianes amb l'efecte de la metformina en càncer de mama triple negatiu i estudiar les diferències d'expressió dels microRNAs que generen resistències al trastuzumab en càncer de mama HER2+, així com estudiar la seua manera de transmissió entre cèl·lules. Es van realitzar assajos cel·lulars tractant amb metformina les línies MDA-MB-231, MDA-MB-468 i MCF-7 així com sobreexpressant o inhibint miR-26a i es van mesurar les seues dianes teòriques per qPCR. Per a les línies HER2+ es va realitzar un Affymetrix Genechip miRNA 4.0 microarray comparant línies SKBR-3wt i BT-474wt amb les seues respectives línies amb resistència generada a trastuzumab i HCC-1954 com resistent innata. Es van estudiar els microRNAs més rellevants de l'array en les línies cel·lulars i en pacients i es va comprovar la seua presència a exosomes, així com l'efecte dels exosomes en la transmissió de la resistència. La sobreexpressió de miR-26a resultà en una reducció de la viabilitat cel·lular que es recuperà parcialment en inhibir-la. E2F3, MCL-1, EZH2, MTDH i PTEN foren regulats negativament per miR-26a i la proteïna PTEN també es va reduir en sobreexpressar miR-26a. El tractament amb metformina va reduir la viabilitat de les cèl·lules de càncer de mama, va augmentar l'expressió de miR-26a i va conduir a una reducció en l'expressió de BCL-2, EZH2 i PTEN. La inhibició de miR-26a prevé part de l'efecte en la viabilitat de la metformina i la reducció de l'expressió de PTEN i EZH2. En les línies HER2+, miR-23b-3p i miR-146a-5p foren els principals candidats extrets de l'array. miR-23b-3p va inhibir PTEN significativament en la línia BT-474. miR-146a-5p va augmentar la resistència de les cèl·lules SKBR-3 al trastuzumab i la seua inhibició va reduir la resistència de les SKBR-3r. L'augment de miR-146a-5p en SKBR-3wt va tindre un efecte en cicle cel·lular augmentant la fase S i la G2/M, inhibint l'expressió de CDKN1A i augmentant la de CCNB1. Els exosomes de les SKBR-3 contenien miR-146a-5p, amb majors nivells en els de les resistents (exoR). Els exoR van augmentar la resistència a trastuzumab, la transició epiteli-mesenquimal i la migració en co-cultivar-los amb SKBR-3wt, i l'angiogènesi de les HUVEC. Els nostres resultats suggereixen que l'efecte de la metformina està intervingut per una major expressió de miR-26a i reducció de les seues dianes, PTEN i EHZ2. Per tant, l'ús de metformina al tractament de el càncer de mama constitueix una prometedora potencial teràpia. En HER2+, miR-23b sembla provocar resistència a trastuzumab mitjançant PTEN i miR-146a a través del cicle cel·lular. A més, miR-146a es transmet en exosomes, que són capaços de reduir la sensibilitat al trastuzumab de les cèl·lules sensibles i augmentar la TEM, migració i angiogènesi.

Osteosarcoma is the most common primary malignant tumor, especially in children and adolescents. Circular RNAs (circRNAs) are found to play roles in the progression of osteosarcoma. However, the exact functions of circRNAs in osteosarcoma development still need to be clarified. We obtained differentially expressed circRNAs and miRNAs from a GSE99671 data set (GEO database). The gene co-expression network of ceRNAs and osteosarcoma-related genes was analyzed using the STRING database. qRT-PCR was used to detect the expression of circ-03955 and miR-3662. Transwell assays and flow cytometry were performed to detect phenotypic changes in cell function. A xenograft tumor model was established using BALB/c nude mice. Dual luciferase activity and RNA immunoprecipitation assays were performed to assess the relationship between circ-03955, miR-3662, and metadherin (MTDH). Immunohistochemistry, immunofluorescence, and Western blotting were used to assess protein expression levels. Circ-03955 was significantly upregulated, and miR-3662 was downregulated in osteosarcoma. Circ-03955 silencing inhibited the growth and metastasis of osteosarcoma. Mechanism analysis revealed that circ-03955 could bind to miR-3662, and the latter could target MTDH, leading to its suppressed expression and facilitating epithelial-mesenchymal transition (EMT). All these findings demonstrate that the presence of circ-03955 promotes EMT in osteosarcoma by acting as miR-3662 sponge-mediated MTDH expression.

Cytoplasmic polyadenylation element-binding protein 3 (CPEB3) is a sequence-specific RNA-binding protein. We had reported that CPEB3 is involved in hepatocellular carcinoma (HCC) progression. However, the underlying mechanisms of CPEB3 in HCC remain unclear. In this study, we firstly performed RNA immunoprecipitation to uncover the transcriptome-wide CPEB3-bound mRNAs (CPEB3 binder) in HCC. Bioinformatic analysis indicates that CPEB3 binders are closely related to cancer progression, especially HCC metastasis. Further studies confirmed that metadherin (MTDH) is a direct target of CPEB3. CPEB3 can suppress the translation of MTDH mRNA in vivo and in vitro. Besides, luciferase assay demonstrated that CPEB3 interacted with 3′-untranslated region of MTDH mRNA and inhibited its translation. Subsequently, CPEB3 inhibited the epithelial–mesenchymal transition and metastasis of HCC cells through post-transcriptional regulation of MTDH. In addition, cpeb3 knockout mice are more susceptible to carcinogen-induced hepatocarcinogenesis and subsequent lung metastasis. Our results also indicated that CPEB3 was a good prognosis marker, which is downregulated in HCC tissue. In conclusion, our results demonstrated that CPEB3 played an important role in HCC progression and targeting CPEB3-mediated mRNA translation might be a favorable therapeutic approach.

Purpose Invasive micropapillary carcinoma (IMPC) of the breast has a high propensity for lymphovascular invasion and axillary lymph node metastasis and displays an ‘inside-out’ growth pattern, but the molecular mechanism of metastasis and tumor cell polarity reversal in IMPC is unclear. Methods and Patients: Luciferase reporter assays and western blotting were performed to confirm that the MTDH 3’UTR bound to miR-30c. Growth curves, tumor sphere formation assays, Transwell migration and invasion assays, mouse xenograft model and immunofluorescence were evaluated to investigate the effects of miR-30c and MTDH. MiRNA in situ hybridization (ISH) and immunohistochemistry (IHC) were carried out on IMPC patient tissues for miR-30c and MTDH expression, respectively. Results We found that miR-30c could directly target the MTDH (metadherin) 3’UTR. MiR-30c overexpression inhibited breast cancer cell proliferation, invasion and metastasis by targeting MTDH in vitro and in vivo. Moreover, miR-30c regulated IMPC cell polarity reversal by targeting MTDH. By in situ hybridization and immunohistochemistry analyses, miR-30c and MTDH were significantly correlated with tumor size, lymph nodule status and tumor grade, the ‘inside-out’ growth pattern, overall survival (OS) and disease-free survival (DFS) in IMPC patients (p

(1) Background: Tumor hypoxia leads to metastasis and certain immune responses, and interferes with normal biological functions. It also affects glucose intake, down-regulates oxidative phosphorylation, and inhibits fatty-acid desaturation regulated by hypoxia-inducible factor 1&alpha, (HIF-1&alpha, ). Although tumor hypoxia has been found to promote tumor metastasis, the roles of HIF-1&alpha, regulated genes and their application are not completely integrated in clinical practice. (2) Methods: We examined the correlation between HIF-1&alpha, metadherin (MTDH), and interleukin (IL)-10 mRNA expression, as well as their expression patterns in the prognosis of breast cancer using the Gene Expression Profiling Interactive Analysis (GEPIA) databases via a web interface, tissue microarrays (TMAs) were stained for MTDH and IL-10 protein expression using immunohistochemistry. (3) Results: HIF-1&alpha, MTDH, and IL-10 mRNA expression are highly correlated and strongly associated with poor prognosis. MTDH and IL-10 protein expression of breast cancer patients usually harbored negative estrogen receptor (ER) or progesterone receptor (PR) status, and late-stage tumors have higher IL-10 expression. With regard to MTDH and IL-10 protein expression status for using univariate and multivariate analysis, the results showed that the protein expression of MTDH and IL-10 in ER-negative or PR-negative breast cancer patients have the worse prognosis. (4) Conclusions: we propose a new insight into hypoxia tumors in the metabolism and immune evidence for breast cancer therapy.

Background The well-known traditional Chinese herbal formula Guizhi Fuling Wan (GFW) was recently reported to improve the curative effects of chemotherapy for ovarian cancer with few clinical side effects. The present study aimed to investigate the reversal mechanism of sera derived from rats exposed to Guizhi Fuling Wan extract (GFWE) in cisplatin-resistant human ovarian cancer SKOV3/DDP cells; the proteins examined included phosphatase and tensin homolog (PTEN) and metadherin (MTDH), and the possible protein interaction between PTEN and MTDH was explored. Methods GFWE was administered to healthy Wistar rats, and the sera were collected after five days. The PubMed and CNKI databases were searched for literature on the bioactive blood components in the sera. The systemsDock website was used to predict potential PTEN/MTDH interactions with the compounds. RT-qPCR, western blotting, and immunofluorescence analyses were used to analyze the mRNA and protein levels of MTDH and PTEN. Laser confocal microscopy and coimmunoprecipitation (co-IP) were used to analyze the colocalization and interaction between MTDH and PTEN. Results Sixteen bioactive compounds were identified in GFWE sera after searching the PubMed and CNKI databases. The systemsDock website predicted the potential PTEN/MTDH interactions with the compounds. RT-qPCR, western blotting, and immunofluorescence analyses showed decreased MTDH expression and increased PTEN expression in the sera. Laser confocal microscopy images and coimmunoprecipitation (co-IP) analyses demonstrated that a colocalization and interaction occurred between MTDH and PTEN, and the addition of the sera changed the interaction status. Conclusions GFWE restored sensitivity to cisplatin by inhibiting MTDH expression, inducing PTEN expression, and improving the interaction between MTDH and PTEN in SKOV3/DDP cells, and these proteins and their interaction may serve as potential targets for cancer treatment. The sera may represent a new source of anticancer compounds that could help to manage chemoresistance more efficiently and safely.

Ovarian cancer (OC) has a high mortality rate among women worldwide. However, even with the advances in detection and therapeutics, the number of cases is increasing worldwide. Increasingly, microRNAs (miRNAs), including miR-497-5p, have been implicated in the progression of many cancers, but the role of miR-497-5p in OC remains unknown. The purpose of this study was to investigate the underlying molecular mechanism of miR-497-5p in OC. Herein, we find that miR-497-5p is down-regulated in OC tissues, and overexpression of miR-497-5p enhances apoptosis in OC cells. The increased apoptosis was correlated with enhanced expression of apoptosis-related proteins. MiR-497-5p directly bound the 3’-untranslated region of metadherin (MTDH), leading to the reduction of MTDH in mRNA and protein levels. Moreover, MTDH knockout promoted the apoptosis of OC cells. Taken together, we conclude that miR-497-5p contributes to cell apoptosis in OC by regulating MTDH.

Resistance to chemotherapy and radiation therapy is considered a major therapeutic barrier in breast cancer. Cancer stem cells (CSCs) play a prominent role in chemo and radiotherapy resistance. The established chemo and radio-resistant triple-negative breast cancer (TNBC) cell line MDA-MB-231/IR displays greater CSC characteristics than the parental MDA-MB-231 cells. Escalating evidence demonstrates that metadherin (MTDH) is associated with a number of cancer signaling pathways as well as breast cancer therapy resistance, making it an attractive therapeutic target. Kaplan&ndash, Meier plot analysis revealed a correlation between higher levels of MTDH and shorter lifetimes in breast cancer and TNBC patients. Moreover, there was a positive correlation between the MTDH and CD44 expression levels in The Cancer Genome Atlas breast cancer database. We demonstrate that MTDH plays a pivotal role in the regulation of stemness in MDA-MB-231/IR cells. Knockdown of MTDH in MDA-MB-231/IR cells resulted in a reduction in the CSC population, aldehyde dehydrogenase activity, and major CSC markers, including &beta, catenin, CD44+, and Slug. In addition, MTDH knockdown increased reactive oxygen species (ROS) levels in MDA-MB-231/IR cells. We found that phenethyl isothiocyanate (PEITC), a well-known pro-oxidant phytochemical, suppressed stemness in MDA-MB-231/IR cells through ROS modulation via the downregulation of MTDH. Co-treatment of PEITC and N-Acetylcysteine (a ROS scavenger) caused alterations in PEITC induced cell death and CSC markers. Moreover, PEITC regulated MTDH expression at the post-transcriptional level, which was confirmed using cycloheximide, a protein synthesis inhibitor.

Background Breast cancer, the most common cancer in women worldwide, causes the vast majority of cancer-related deaths. Undoubtedly, tumor metastasis and recurrence are responsible for more than 90 percent of these deaths. MicroRNAs are endogenous noncoding RNAs that have been integrated into almost all the physiological and pathological processes, including metastasis. In the present study, the role of miR-128 in breast cancer was investigated. Results Compared to the corresponding adjacent normal tissue, the expression of miR-128 was significantly suppressed in human breast cancer specimens. More importantly, its expression level was reversely correlated to histological grade of the cancer. Ectopic expression of miR-128 in the aggressive breast cancer cell line MDA-MB-231 could inhibit cell motility and invasive capacity remarkably. Afterwards, Metadherin (MTDH), also known as AEG-1 (Astrocyte Elevated Gene 1) and Lyric that implicated in various aspects of cancer progression and metastasis, was further identified as a direct target gene of miR-128 and its expression level was up-regulated in clinical samples as expected. Moreover, knockdown of MTDH in MDA-MB-231 cells obviously impaired the migration and invasion capabilities, whereas re-expression of MTDH abrogated the suppressive effect caused by miR-128. Conclusions Overall, these findings demonstrate that miR-128 could serve as a novel biomarker for breast cancer metastasis and a potent target for treatment in the future.

Background: Metadherin (MTDH) has been known as an essential oncogene in carcinogenesis and tumor spread in several malignancies, via its effect on pathways of signal transduction. Objective: We aimed to evaluate the role of serum MTDH mRNA expression in the diagnosis of hepatocellular carcinoma (HCC) and to compare its expression levels with serum levels of Alpha-fetoprotein (AFP). Subjects & methods: A total of 150 subjects (90 HCC patients & 60 healthy volunteers) were enrolled in the current study. Serum MTDH mRNA relative expression was analyzed by Real Time PCR technique. Results: There was a significant statistical increase of serum MTDH mRNA expression in HCC group when compared to controls (P < 0.05). MTDH mRNA expression was significantly associated with clinicopathological data, advanced tumor stage and poor histological differentiation in HCC patients (p < 0.05). There was direct positive correlation between MTDH mRNA expression and serum AFP levels in HCC group (r = 0.445), P value =

In this study, we evaluated the function and regulation of the long non-coding RNA (lncRNA) FAM83H-AS1 in triple-negative breast cancer (TNBC). Our data show that the FAM83H-AS1 levels are increased in human TNBC cells and tissues. Proliferation, migration, and invasion of TNBC cells are decreased by FAM83H-AS1 suppression, but increased by FAM83H-AS1 overexpression. Bioinformatics analysis revealed that miR-136-5p is a potential target of FAM83H-AS1. MiR-136-5p expression is decreased in TNBC tissues, and its overexpression suppresses TNBC cell proliferation, migration, and invasion. MiR-136-5p suppression reverses the FAM83H-AS1 silencing-mediated inhibition of TNBC cell proliferation, migration, and invasion, suggesting that FAM83H-AS1 exerts its oncogenic effect by inhibiting miR-136-5p. Our data identify metadherin (MTDH) as the target gene of miR-136-5p, and demonstrate that the MTDH expression is increased in human TNBC tissues, which induces proliferation, migration, and invasion of TNBC cells. Importantly, our in vivo data show that FAM83H-AS1 also promotes tumor growth in TNBC mouse xenografts. Together, our results demonstrate that FAM83H-AS1 functions as an oncogenic lncRNA that regulates miR-136-5p and MTDH expression during TNBC progression, and suggest that targeting the FAM83H-AS1/miR-136-5p/MTDH axis may serve as a novel therapeutic target in TNBC.

Malignant glioma is the most common and lethal type of primary tumor of the central nervous system. The incidence of glioma is increasing year by year. In recent years, a variety of new treatment methods have emerged, among which gene therapy has become a hotspot. MicroRNAs (miRNAs) are a class of small non-coding single-strand RNAs that negatively regulate gene expression at the post-transcriptional and/or translational level by binding loosely complimentary sequences in the 3′ untranslated regions (UTRs) of target mRNAs. Several miRNAs have been reported to modulate glioma progression. This study aimed to determine the function of miR-30b-5p in glioma and its underlying molecular mechanism. miR-30b-5p expression was significantly lower in gliomas than the normal brain tissues. Overexpression of miR-30b-5p was found to significantly inhibit glioma cell proliferation in vitro. Further, MTDH expression was significantly higher in the gliomas compared with the normal brain tissues. In addition, MTDH was validated as direct target of miR-30b-5p. Moreover, cellular proliferation was increased after MTDH overexpression in the glioma cells, which reversed the effects of miR-30b-5p. Taken together, these results reveal miR-30b-5p impacts glioma cell proliferation via direct targeting MTDH and could be a potential novel therapeutic target for the treatment of glioma. Keywords: miR-30b-5p, MTDH, Proliferation

Objective Chromosomal alterations and copy number changes are frequent events in tumors, leading to amplification of focal regions containing several oncogenes. Gains and losses of several regions have been reported in head and neck cancer (HNC) but the copy number changes of the individual genes located in these regions have not been analyzed so far. In this study we aimed to analyze the copy number variations in patients with HNC. Design Prospective study Setting University hospital Participants 50 patients with squamous cell carcinoma of the head and neck Methods Copy number changes and amplifications of 22 genes in tumors and matched tissue were analyzed by MLPA which allows simultaneous analysis of gene copy numbers in multiple genetic regions. Results Amplifications were observed in 52% and losses were detected in 20% of the samples. Chromosome 8 was found to harbor the most frequent copy number alterations. The most frequently amplified genes were CCND1 and the MED1 genes followed by the MTDH and MYC genes on the long arm and ZNF703 on the short arm of chromosome 8. Amplification of the ZNF703, PRDM14 and MYC genes were highly correlated suggesting that the genes displaying high copy number changes on chromosome 8 collaborate during carcinogenesis. Conclusions The alterations found in our study supports the contribution of gene amplifications and indicate cooperation between certain oncogenes in the pathogenesis of HNSCC. Correlations between amplification of less familiar genes and known oncogenes warrant further investigation. This article is protected by copyright. All rights reserved.

ObjectiveReliable biomarkers associated with aggressiveness of non-functioning gonadotroph adenomas (GAs) are lacking. As the growth of tumor remnants is highly variable, molecular markers for growth potential prediction are necessary. We hypothesized that fast- and slow-growing GAs present different gene expression profiles and reliable biomarkers for tumor growth potential could be identified, focusing on the specific role of epithelial-mesenchymal transition (EMT).Design and methodsEight GAs selected for RNA sequencing were equally divided into fast- and slow-growing group by the tumor volume doubling time (TVDT) median (27.75 months). Data were analyzed by tophat2, cufflinks and cummeRbund pipeline. 40 genes were selected for RT-qPCR validation in 20 GAs based on significance, fold-change and pathway analyses. The effect of silencingMTDH(metadherin) andEMCN(endomucin) onin vitromigration of human adenoma cells was evaluated.Results350 genes were significantly differentially expressed (282 genes upregulated and 68 downregulated in the fast group,P-adjusted RP PCDH18, UNC5D, EMCN, MYO1B, GPM6Aand six EMT-related genes (SPAG9, SKIL, MTDH, HOOK1, CNOT6LandPRKACB).MTDH, but notEMCN, demonstrated involvement in cell migration and association with EMT markers.ConclusionsFast- and slow-growing GAs present different gene expression profiles, and genes related to EMT have higher expression in fast-growing tumors. In addition toMTDH, identified as an important contributor to aggressiveness, the other genes might represent markers for tumor growth potential and possible targets for drug therapy.