Search

Formononetin-inhibited IgE-mediated mast cell activation and attenuated IgE/Ag-induced allergic inflammation by suppressing USP5 or USP13 expression and increasing proteasome-mediated FcεRIγ degradation.

This study aimed to determine whether the prevalence of thyroid nodules (TNs) increased due to modern lifestyles or other factors, despite the advances in screening and diagnostic tools.This study included 3474 pairs of participants, who were matched by gender and age (±3 years) from two cross-sectional sampling surveys: (1) the program on the iodine nutritional status and related health status of residents in Shanghai in 2009; (2) the thyroid disease screening program for adults in Shanghai between 2017 and 2018. The prevalence of TNs and thyroid diseases in 2009 and 2017-2018 were compared, and the potential risk factors of TNs were detected.The prevalence of TNs in 2009 was 28.9%: 22.5% in males and 34.5% in females. In 2017, this increased to 43.8%: 37.9% in males and 49.1% in females. The prevalence of TNs significantly increased from 2009 to 2017 (odds ratio, 1.486; 95% confidence interval, 1.238-1.786). In addition, female gender, thyroid disease history, and age were the main risk factors for TNs after adjusting for confounders in the logistic regression across the time period.The prevalence of TNs significantly increased across nearly 10 years in Shanghai.

The third-generation epidermal growth factor receptor (EGFR) inhibitor osimertinib (OSI) has been approved as the first-line treatment for EGFR-mutant non-small cell lung cancer (NSCLC). This study aims to explore a rational combination strategy for enhancing the OSI efficacy. In this study, OSI induced higher CD47 expression, an important anti-phagocytic immune checkpoint, via the NF-κB pathway in EGFR-mutant NSCLC HCC827 and NCI-H1975 cells. The combination treatment of OSI and the anti-CD47 antibody exhibited dramatically increasing phagocytosis in HCC827 and NCI-H1975 cells, which highly relied on the antibody-dependent cellular phagocytosis effect. Consistently, the enhanced phagocytosis index from combination treatment was reversed in CD47 knockout HCC827 cells. Meanwhile, combining the anti-CD47 antibody significantly augmented the anticancer effect of OSI in HCC827 xenograft mice model. Notably, OSI induced the surface exposure of "eat me" signal calreticulin and reduced the expression of immune-inhibitory receptor PD-L1 in cancer cells, which might contribute to the increased phagocytosis on cancer cells pretreated with OSI. In summary, these findings suggest the multidimensional regulation by OSI and encourage the further exploration of combining anti-CD47 antibody with OSI as a new strategy to enhance the anticancer efficacy in EGFR-mutant NSCLC with CD47 activation induced by OSI.

Background Biliary atresia (BA) is one of the main causes of neonatal end-stage liver disease. Without timely diagnosis and treatment, most children with BA will develop irreversible liver fibrosis within the first two months. While current theorized causes of BA include viral infection, immune disorders, and genetic defects, the comprehensive etiology is still largely unknown. Recently, biliatresone attracted much interest for its ability to induce BA in both zebrafish and mice, so we summarized the latest progress of biliatresone research in BA and tried to answer the question of whether it could provide further clues to the etiology of human BA. Data sources We conducted a PubMed search for any published articles related to the topic using search terms including “biliary atresia”, “biliatresone”, “GSH”, and “HSP90”. Relevant data were extracted from the original text or supplementary materials of the corresponding articles. Results Biliatresone had shown its unique toxicity in multiple species such as zebrafish and mice, and pathogenic factors involved included glutathione (GSH), heat shock protein 90 (HSP90) and the related pathways. In combination with epidemiological evidence and recent studies on the intestinal flora in biliary atresia, a new pathogenic hypothesis that the occurrence of biliary atresia is partly due to biliatresone or its structure-like compounds depositing in human body via vegetables or/and the altered intestinal flora structure can be tentatively established. Conclusions Based on the existing evidence, we emphasized that GSH and HSP90 are involved in the development of BA, and the maternal diet, especially higher vegetable intake of Asian women of childbearing age, accompanied by the altered intestinal flora structure, may contribute to the occurrence of biliary atresia and the higher incidence in the Asia group. However, the evidence from large sample epidemiological research is necessary.

Nitrogen is an essential element required for plant growth and productivity. Understanding the mechanisms and natural genetic variation underlying nitrogen use in plants will facilitate the engineering of plant nitrogen use to maximize crop productivity while minimizing environmental costs. To understand the scope of natural variation that may influence nitrogen use, we grew 1,135 Arabidopsis thaliana natural genotypes on two nitrogen sources, nitrate and ammonium, and measured both developmental and defense metabolite traits. By using different environments and focusing on multiple traits, we identified a wide array of different nitrogen responses. These responses are associated with numerous genes, most of which were not previously associated with nitrogen responses. Only a small portion of these genes appear to be shared between environments or traits, while most are predominantly specific to a developmental or defense trait under a specific nitrogen source. Finally, by using a large population, we were able to identify unique nitrogen responses, such as preferring ammonium or nitrate, which appear to be generated by combinations of loci rather than a few large-effect loci. This suggests that it may be possible to obtain novel phenotypes in complex nitrogen responses by manipulating sets of genes with small effects rather than solely focusing on large-effect single gene manipulations.

Programmed death ligand 1 (PD-L1) is widely known as an immune checkpoint, and immunotherapy through the inhibition of checkpoint molecules has become an important component in the successful treatment of tumours via programmed death 1 (PD-1)/PD-L1 signalling pathways. However, its biological functions and expression profile in colorectal cancer (CRC) are elusive. We previously found that PD-L1 can bind to PD-L1 and cause cell detachment. However, the detailed molecular mechanisms of how PD-L1 binds to PD-L1 and how it transmits signals to the cell remain unclear. In this study, we disclosed that PD-L1 expression was dramatically upregulated in CRC compared to normal tissues. Ectopic expression of PD-L1 inhibits cell adhesive capacity and promotes cell migration in CRC cell lines, while silencing PD-L1 had the opposite effects and suppressed invasion and proliferation. Mechanistically, PD-L1 was found to promote epithelial-mesenchymal transition (EMT) through the ERK signalling molecule pathway and interacted with the 1-86 aa fragment of KRAS to transduce signals. Collectively, our study demonstrated the role of PD-L1 after binding to PD-L1 in CRC, thereby providing a new theoretical basis for further improving immunotherapy with anti-PD-L1 antibodies.

Varicosporellopsis shangrilaensis Z. R. Dou & K. L. Wang, sp. nov. (Figure 2) Index Fungorum number: IF 559862 Etymology: —The epithet “ shangrilaensis ” refers to the name of the host location, Shangri-la, where the holotype was collected. Holotype: — CHINA, Yunnan Province, Diqing City, Shangri-la County, N 27°26′19.715″ E 99°50′24.193″, from the rhizosphere soil of the plant Astragalus polycladus Bur. et Franch; 24 September 2021, Z. R. Dou, CGMCC 3.21000 holotype, deposited in the China General Microbiological Culture Collection Center. Ex-type living culture MTF 7, preserved in the Dali University Culture Collection. Description:—Asexual morph colonies on PDA initially white, gradually turning to pale yellow after two weeks, cottony, growing slowly, reaching 30 mm diam, after 10 days at 26 °C. Mycelium hyaline, branched, smooth, consisting of septate, frequently forming hyphal coils. Conidiophores macronematous, erect, septate, unbranched or branched at the base. Conidiogenous cells 10–20 × 3–5 μm (x̅ = 14.5 × 4.3 μm, n = 100), phialidic, with a minute collarette. Conidia two types: Macroconidia 8–20 × 2.5–8.5 μm (x̅ = 13.5 × 4.5 μm, n = 100), reniform or cymbiform, straight or slightly curved, guttulate, aseptate, hyaline, smooth; Microconidia 3.5–6.5 × 3–6 μm (x̅ = 4.9 × 4.8 μm, n = 100), spherical or teardrop-shaped, guttulate, aseptate, hyaline, smooth. Chlamydospores not observed. Sexual morph was undetermined. Known distribution: — Diqing City (H about 3,226 m), Shangri-la County, Yunnan Province, China. Additional material examined: — CHINA, Yunnan Province, Diqing City, Shangri-la County, N 27°26′19.715″ E 99°50′24.193″, from rhizosphere soil of Astragalus polycladus, 24 September 2021, K. L. Wang and Z. R. Dou, living culture KLF 01.

Varicosporellopsis shangrilaensis Z. R. Dou & K. L. Wang, sp. nov. (Figure 2) Index Fungorum number: IF 559862 Etymology: —The epithet “ shangrilaensis ” refers to the name of the host location, Shangri-la, where the holotype was collected. Holotype: — CHINA, Yunnan Province, Diqing City, Shangri-la County, N 27°26′19.715″ E 99°50′24.193″, from the rhizosphere soil of the plant Astragalus polycladus Bur. et Franch; 24 September 2021, Z. R. Dou, CGMCC 3.21000 holotype, deposited in the China General Microbiological Culture Collection Center. Ex-type living culture MTF 7, preserved in the Dali University Culture Collection. Description:—Asexual morph colonies on PDA initially white, gradually turning to pale yellow after two weeks, cottony, growing slowly, reaching 30 mm diam, after 10 days at 26 °C. Mycelium hyaline, branched, smooth, consisting of septate, frequently forming hyphal coils. Conidiophores macronematous, erect, septate, unbranched or branched at the base. Conidiogenous cells 10–20 × 3–5 μm (x̅ = 14.5 × 4.3 μm, n = 100), phialidic, with a minute collarette. Conidia two types: Macroconidia 8–20 × 2.5–8.5 μm (x̅ = 13.5 × 4.5 μm, n = 100), reniform or cymbiform, straight or slightly curved, guttulate, aseptate, hyaline, smooth; Microconidia 3.5–6.5 × 3–6 μm (x̅ = 4.9 × 4.8 μm, n = 100), spherical or teardrop-shaped, guttulate, aseptate, hyaline, smooth. Chlamydospores not observed. Sexual morph was undetermined. Known distribution: — Diqing City (H about 3,226 m), Shangri-la County, Yunnan Province, China. Additional material examined: — CHINA, Yunnan Province, Diqing City, Shangri-la County, N 27°26′19.715″ E 99°50′24.193″, from rhizosphere soil of Astragalus polycladus, 24 September 2021, K. L. Wang and Z. R. Dou, living culture KLF 01., Published as part of Dou, Zheng-Rong, Zhang, Fa, Zhou, Guo-Jun, Gong, Gao-Fen, Ding, Jia-Jie, Fan, Xu-Hang, Jiang, Bei & Wang, Kai-Ling, 2023, Varicosporellopsis shangrilaensis sp. nov. (Nectriaceae, Hypocreales), a new terricolous species isolated from the rhizosphere soil of Astragalus polycladus in northwestern Yunnan, China, pp. 230-238 in Phytotaxa 600 (4) on pages 233-234, DOI: 10.11646/phytotaxa.600.4.2, http://zenodo.org/record/8093963

Dou, Zheng-Rong, Zhang, Fa, Zhou, Guo-Jun, Gong, Gao-Fen, Ding, Jia-Jie, Fan, Xu-Hang, Jiang, Bei, Wang, Kai-Ling (2023): Varicosporellopsis shangrilaensis sp. nov. (Nectriaceae, Hypocreales), a new terricolous species isolated from the rhizosphere soil of Astragalus polycladus in northwestern Yunnan, China. Phytotaxa 600 (4): 230-238, DOI: 10.11646/phytotaxa.600.4.2, URL: http://dx.doi.org/10.1094/PDIS-04-22-0755-PDN

YAG (Y3Al5O12) transparent ceramics have attractive application prospects for transparent armor protection modules because of their excellent light transmittance and anti-ballistic capability. Understanding the fracture behavior and damage mechanism of YAG is necessary for armor design. To explore the damage characteristics of YAG under compression and tension, shock compression and shockless spalling experiments with soft recovery technique are conducted. The spall strength of YAG is obtained and the recovered samples are observed by CT and SEM. It is shown that the macroscopic damage characteristic of YAG under compression is vertical split cracks with oblique fine cracks distributed in the entire sample, while that under tension is horizontal transgranular cracks concentrated near the main spall surface. The cracks generated by macroscopic compression, tension and shear stress extend in similar tensile form at the microscale. The proportion of transgranular fractures on spall surfaces is higher than that of cracks induced by macroscopic compression. Meanwhile, higher loading rate and longer loading duration increase the transgranular fracture percentage.

To evaluate the clinical characteristics, biochemical parameters and the distribution of HLA-DQ genotypes among adult patients with celiac disease (CD) in Northwest China.This cross-sectional study retrospectively collected clinical, biochemical, and HLA-DQ genotype of patients with CD from a tertiary hospital in Xinjiang Uygur Autonomous Region, China between March 2016 and December 2021. Small intestinal biopsy and serum-specific antibodies were used to diagnose CD.Of the 102 CD patients, 63.7% were women (female: male = 1.76:1), and the mean age was 47.3 ± 14.7 years at diagnosis. Common gastrointestinal symptoms included abdominal pain (50.0%), diarrhea (39.2%), and abdominal distension (24.5%). While common extraintestinal manifestations were anemia (48.0%), osteopenia or osteoporosis (36.3%), and fatigue (35.3%). Approximately 34.3% of patients with CD had comorbidities, with the most common being thyroid diseases (18.6%). Biochemical profiles showed lower hemoglobin, higher platelet count, and 25-hydroxyvitamin D (25[OH]D) deficiency. HLA-DQ2/DQ8 was detected among all 53 patients who underwent genotype testing; the frequency of the HLA-DQ2.5, DQ2.2, and DQ8 haplotypes was 71.7%, 24.5%, and 3.8%, respectively.CD was more common among women. Clinical manifestations include primarily gastrointestinal symptoms, but extraintestinal manifestations were not uncommon. Lower hemoglobin level, higher platelet count, and 25[OH]D deficiency are the main biochemical manifestations. The HLA-DQ2.5 and DQ2.2 haplotypes are the most common genotypes in CD.

Imaging examinations following sublobar resection of lung cancer often find thickened neoplasms around the resection margin. Identifying whether the neoplasms are postoperative recurrence or margin granulomas is vitally important. However, the identification mainly depends on the empirical judgment of the imaging department or clinicians in each clinical center at present, and there are few relevant studies, so it is hard to formulate a relatively unified standard. Therefore, we collected data from patients with thickened neoplasms around the resection margin after sublobar resection and sought to discover how to differentiate granulomas from tumor recurrence.We examined the clinical records of 15 patients with neoplasms around the margins which identified as malignant in auxiliary examination reports, and received second surgery after first sublobar resection. We collected their postoperative pathology and auxiliary examination parameters. The univariate predictors helpful in distinguishing between recurrence and granuloma were analyzed as a diagnostic test.Of the 15 patients with neoplasms around the resection margin, six were diagnosed with benign granulomas, and nine were diagnosed with primary lung cancer recurrence. The results revealed that age, gender, specific surgical method, maximum standardized fluorodeoxyglucose uptake value (SUVmax), and follow-up time were not significantly different, but there were significant differences in enhanced computed tomography (CT) values in several analyses, which calculated by the hospital imaging system. The maximum CT values of the tumor recurrence and granuloma were 104.9±8.051 and 130.3±7.017 (P=0.045), the minimum CT values (15.67±5.113Differentiating between tumor recurrence and granulomas after sublobar resection remains difficult. However, examining the differences in enhanced CT allows the clinician to make an informed diagnosis that aids further investigation and treatment.

Peritoneal dialysis (PD) is a widely accepted renal replacement therapy for patients with end-stage renal disease (ESRD). Morphological and functional changes occur in the peritoneal membranes (PMs) of patients undergoing long-term PD. Peritoneal fibrosis (PF) is a common PD-related complication that ultimately leads to PM injury and peritoneal ultrafiltration failure. Autophagy is a cellular process of “self-eating” wherein damaged organelles, protein aggregates, and pathogenic microbes are degraded to maintain intracellular environment homeostasis and cell survival. Growing evidence shows that autophagy is involved in fibrosis progression, including renal fibrosis and hepatic fibrosis, in various organs. Multiple risk factors, including high-glucose peritoneal dialysis solution (HGPDS), stimulate the activation of autophagy, which participates in PF progression, in human peritoneal mesothelial cells (HPMCs). Nevertheless, the underlying roles and mechanisms of autophagy in PF progression remain unclear. In this review, we discuss the key roles and potential mechanisms of autophagy in PF to offer novel perspectives on future therapy strategies for PF and their limitations.

Studies using rodent models have shown that relapse to drug or food seeking increases progressively during abstinence, a phenomenon termed ‘incubation of craving’. Mechanistic studies of incubation of craving have focused on specific neurobiological targets within pre- selected brain areas. Recent methodological advances in whole-brain immunohistochemistry, clearing, and imaging now enable unbiased brain-wide cellular resolution mapping of regions and circuits engaged during learned behaviors. However, these whole brain imaging approaches were developed for mouse brains while incubation of drug craving has primarily been studied in rats and incubation of food craving has not been demonstrated in mice. Here, we established a mouse model of incubation of palatable food craving and examined food reward seeking after 1, 15, and 60 abstinence days. We then used the neuronal activity marker Fos with intact brain mapping procedures to identify corresponding patterns of brain-wide activation. Relapse to food seeking was significantly higher after 60 abstinence days than after 1 or 15 days. Using unbiased ClearMap analysis, we identified increased activation of multiple brain regions, particularly corticostriatal structures, following 60, but not 15 abstinence days. We used orthogonal SMART2 analysis to confirm these findings within corticostriatal and thalamocortical subvolumes and applied expert-guided registration to investigate subdivision and layer-specific activation patterns. Overall, we (1) identified novel brain-wide activity patterns during incubation of food seeking using complementary analytical approaches, and (2) provide a single-cell resolution whole-brain atlas that can be used to identify functional networks and global architecture underlying incubation of food craving.Significance StatementRelapse to reward seeking progressively increases during abstinence, a phenomenon termed incubation of craving. Mechanistic studies of incubation can lead to novel relapse treatments. However, previous studies have primarily used rat models and targeted region-by-region analyses and a brain-wide functional atlas of incubation of reward seeking is lacking. We established a behavioral procedure for incubation of palatable food seeking in mice and applied whole-brain activity mapping with Fos as a neuronal activity marker to identify the functional connectome of this incubation. Like rats, mice showed incubation of food seeking during abstinence. Using two complementary activity mapping approaches, we identified a brain-wide pattern of increased neural activation that mirrored incubation of food seeking after 60, but not 15, days of abstinence.

Background The detachment of the left atrial appendage(LAA) thrombus is the major cause of cardioembolic stroke(CES)/cardioembolic transient ischemic attack(CETIA). A LAA thrombus is very infrequently detected in the presence of sinus rhythm. Patent foramen ovale(PFO) is another common reason for CES/CETIA, especially among young and middle-aged people.Case Presentation We reported a case of a 46-year-old man who presented with TIAs and was subsequently found with PFO and LAA thrombus despite being in sinus rhythm and basically normal structure and function. Anticoagulant treatment was effective, and there was no recurrence of TIA or stroke.Conclusion LAA thrombus could occur in sinus rhythm and normal structure and function. The PFO may be an innocent bystander in the patient suspected of CES/CETIA. Percutaneous PFO closure should be cautiously chosen.

Topological insulators and semimetals have exotic surface and bulk states with massless Dirac or Weyl fermions, demonstrating microscopic transport phenomenon based on relativistic theory. Chiral anomaly induced negative magnetoresistance (negative MR) under parallel magnetic field and current has been used as a probable evidence of Weyl fermions in recent years. Here we report a novel negative MR result with mutually perpendicular in-plane magnetic field and current in Cd3As2 nanowires. The negative MR has a considerable value with -16% around 1.5 K and could persist to room temperature of 300 K with -1%. The gate tuning and angle dependence of the negative MR demonstrate the mechanism of the observed negative MR is different from chiral anomaly. Percolating current paths induced by charge puddles and disorder might be involved to produce such considerable negative MR. Our results indicate the negative MR effect in topological semimetals involves synergistic effects of many mechanisms despite chiral anomaly.

PDF file - 82K

PDF file - 1061K, S Figure 1. Down-regulation of CRT expression reduces PTP1B mRNA and protein expression in KYSE510 cells. S2. PTP1B promotes ESCC cell invasion and migration in KYSE510 cells. KYSE510 cells were transfected with PTP1B-specific siRNA (siRNA PTP1B) or non-targeting siRNA control (siRNA cont). S3. CRT regulates cell motility through PTP1B in ESCC cells. (A-C) 1x105 KYSE150 cells were seeded on 24-well plates with three replicates. S4. Cancer-inflammatory response was minimal in mouse model. S5. CRT regulates PTP1B transcription through Stat5a. S6. The expression of CRT and PTP1B in 176 ESCC tissues was evaluated by immunohistochemistry. S7. Correlation of PTP1B expression with overall survival in patients with CRT high-expressing tumors. S8. Model of CRT-dependent signaling pathway in ESCC cell metastasis. Supplementary Table 1. The candidate genes altered in CRT-siRNA KYSE150 cells as compared with non-targeting siRNA cells detected by cDNA array

PDF file - 927K, The effect of Aurora kinases inhibitors on HL-60 and KG1a cell lines.

PDF file - 1715K, Aurora kinases inhibitors and mTOR inhibitors had synergistic effect on the expression of cleaved caspase 3 and p62.

PDF file - 1113K, Aurora kinases inhibitors had no effect with mTOR inhibitors on PBMCs.

PDF file - 1100K, Combination of Aurora kinases inhibitors and 2DG had minimal effect on PBMCs.

Calreticulin (CRT) is a Ca2+-binding chaperone protein that alters cellular Ca2+-homeostasis in the endoplasmic reticulum (ER). Previously it was shown that CRT was overexpressed in esophageal squamous cell carcinoma (ESCC), and elevated CRT expression promoted the migration and invasion of ESCC cells. In the present study, the mechanisms underlying the role of CRT in esophageal carcinoma progression were investigated. Critically, depletion of CRT or protein-tyrosine phosphatase 1B (PTP1B) reduced ESCC cell migration and metastasis to the lung, whereas restoration of PTP1B protein levels rescued cell migration in CRT-silenced cells. Knockdown of CRT decreased PTP1B protein expression by reducing phosphorylation at the Y694 site of STAT5A, whereas knockdown of PTP1B reduced ERK1/2 phosphorylation at T204. Immunohistochemical analysis of CRT and PTP1B expression in ESCC patient tissues was strongly correlated. Importantly, PTP1B expression was associated with poor survival in patients with CRT overexpression. Overall, these data indicate a novel signaling pathway connecting CRT, STAT5A, PTP1B, and ERK1/2 in the regulation of ESCC cell migration.Implications: These findings suggest that PTP1B is a downstream effector of CRT signaling, promotes tumor progression, and can potentially be used as a new drug target for ESCC. Mol Cancer Res; 11(9); 986–94. ©2013 AACR.

PDF file - 1470K, Aurora kinases inhibitors had synergistic effect with mTOR inhibitors on the glycolytic metabolism of U937 cells.

PDF file - 2735K, Aurora kinases inhibitors had synergistic effect with mTOR inhibitors on the proliferation of U937 and NB4 cells.

PDF file - 1081K, Cell viability detection in Aurora kinase A knockdown-U937 cells.

Purpose: We previously revealed that the calreticulin (CRT) gene is a candidate oncogene promoting cell migration and invasion and that neuropilin-1 (NRP1) is a possible effector downstream of CRT in esophageal squamous carcinoma cells. This study aims to explore the mechanisms underlying the migration and invasion of esophageal cancer cells regulated by CRT through NRP1.Experimental Design: Quantitative reverse-transcription polymerase chain reaction, Western blot analysis, chromatin immunoprecipitation, and reporter gene assays were used to investigate the relationship between CRT and NRP1. In vitro and in vivo assays were carried out to evaluate the effects of NRP1 on malignant phenotypes of ESCC cells and tumor metastasis in NOD/SCID mice. Immunohistochemistry was performed to analyze the expression of CRT and NRP1 in esophageal squamous cell carcinomas (ESCC).Results: Knockdown of CRT decreased the expression of NRP1. Inhibition of NRP1 reduced ESCC cell motility in vitro and experimental metastasis in vivo. Ectopic expression of NRP1 rescued the defects of cell migration and invasion in CRT-shRNA cells. CRT depletion inhibited STAT5A phosphorylation at the Y694 site via a CaMKII-independent pathway. Moreover, STAT5A directly regulated NRP1 transcription. Knockdown of CRT or NRP1 led to a downregulation of MMP2, MMP9, and FAK. Notably, positive correlation was found between CRT and NRP1 expression in ESCC tissues (P = 5.87 × 10−5). CRT and NRP1 coexpression was significantly associated with lymph node metastasis (P = 0.025).Conclusions: Our findings suggest that NRP1 is a critical downstream effector of CRT in promoting cell migration and invasion, which might contribute to the metastasis of ESCC. Clin Cancer Res; 20(23); 6153–62. ©2014 AACR.

Effect of ANO1 on cell proliferation. Knock-down the expression of ANO1 inhibited the cell proliferation of KYSE30 (A) and KYSE510 cells (B). The absorbance at 450 nm was measured.

Supplementary Table 2. Relationship between NRP1/CRT co-expression and lymph node metastasis in 216 ESCC patients

Supplementary Figure 2. Quantitative real-time PCR analysis of STAT5A and STAT5B expression in ESCC cells transiently transfected with non-silencing siRNA, STAT5A siRNA or STAT5B siRNA. Columns, mean; Bars, SD (n = 3); **, P < 0.01.

Amplification and overexpression of MCM7 in ESCC. A. Amplification of MCM7 in array CGH study. B. Validation of MCM7 amplification by FISH. Left: two MCM7 signals in peripheral blood cells; Middle: two MCM7 signals in ESCC cells; Right: more than four MCM7 signals in ESCC cells. C. MCM7 overexpressed in ESCC by semi-quantitative RT-PCR. D. MCM7 overexpressed in ESCC by Western blotting. E. MCM7 overexpressed in ESCC by IHC.

charateritics of 84 HNSCC patients

Supplementary Data. Supplementary Materials and Methods, Supplementary Figure Legends

S1. Identification of RARS-MAD1L1 by RNA-Seq. S2. RARS-MAD1L1 promotes the proliferation and colony-formation ability of p53-/- MEFs and S26 cells. S3. RARS-MAD1L1 enhances CSC properties in NPC cells. S4. Knockdown of endogenous RARS-MAD1L1 reduces the CSC characteristics of C666-1 cells. S5. Knockdown of RARS-MAD1L1 reduces the oncogenic function of S26 cells induced by RARS-MAD1L1. S6. RARS-MAD1L1 overexpression presents a stronger oncogenic function than wild-type RARS in NPC cells. S7. Knockdown of c-Myc reduces the expression of ABCG2 S8. Schematic illustration of the mechanism through which RARS-MAD1L1 induces cancer stem cell properties and therapeutic resistance through the FBP1/c-Myc axis.

Frequency plot comparison analysis. A. Frequency plot comparison of ESCC with or without lymph node metastasis. B. Frequency plot comparison of ESCC at different stages.

Purpose: Nasopharyngeal carcinoma (NPC) is the most common head and neck cancer in Southeast Asia. Because local recurrence and distant metastasis are still the main causes of NPC treatment failure, it is urgent to identify new tumor markers and therapeutic targets for advanced NPC.Experimental Design: RNA sequencing (RNA-seq) was applied to look for interchromosome translocation in NPC. PCR, FISH, and immunoprecipitation were used to examine the fusion gene expression at RNA, DNA, and protein levels in NPC biopsies. MTT assay, colony formation assay, sphere formation assay, co-immunoprecipitation, chromatin immunoprecipitation assay, and in vivo chemoresistance assay were applied to explore the function of RARS-MAD1L1 in NPC.Results: We demonstrated that RARS-MAD1L1 was present in 10.03% (35/349) primary NPC biopsies and 10.7% (9/84) in head and neck cancer (HNC) samples. RARS-MAD1L1 overexpression increased cell proliferation, colony formation, and tumorigenicity in vitro, and the silencing of endogenous RARS-MAD1L1 reduced cancer cell growth and colony formation in vitro. In addition, RARS-MAD1L1 increased the side population (SP) ratio and induced chemo- and radioresistance. Furthermore RARS-MAD1L1 interacted with AIMP2, which resulted in activation of FUBP1/c-Myc pathway. The silencing of FUBP1 or the administration of a c-Myc inhibitor abrogated the cancer stem cell (CSC)-like characteristics induced by RARS-MAD1L1. The expression of c-Myc and ABCG2 was higher in RARS-MAD1L1–positive HNC samples than in negative samples.Conclusions: Our findings indicate that RARS-MAD1L1 might contribute to tumorigenesis, CSC-like properties, and therapeutic resistance, at least in part, through the FUBP1/c-Myc axis, implying that RARS-MAD1L1 might serve as an attractive target for therapeutic intervention for NPC. Clin Cancer Res; 24(3); 659–73. ©2017 AACR.

Supplemental Methods and Materials

Supplementary Figures S1-S8; Supplementary Tables S1-S4.

RARS-MAD1L1 gDNA fusion sequence by Sanger Sequencing