Search

Clear cell renal cell carcinoma (ccRCC) is one of the most common malignant tumors and is characterized by a poor prognosis. Although G2- and S -phase expressed-1 (GTSE1) is known to be involved in the progression and metastasis of various cancers, its significance and mechanism in ccRCC remain unknown. In the present study, we found that GTSE1 was overexpressed in ccRCC tissues, especially in metastatic samples. Moreover, high GTSE1 expression was positively correlated with higher pT stage, tumor size, clinical stage, and WHO/ISUP grade and worse prognosis. And GTSE1 expression served as an independent prognostic factor for overall survival (OS). In addition, GTSE1 knockdown inhibited ccRCC cell proliferation, migration, and invasion, and enhanced cell apoptosis in vitro and in vivo. GTSE1 was crucial for epithelial-mesenchymal transition (EMT) in ccRCC. Mechanistically, GTSE1 depletion could upregulate the expression of Krüppel-like factor 4 (KLF4), which acts as a tumor suppressor in ccRCC. Downregulation of KLF4 effectively rescued the inhibitory effect induced by GTSE1 knockdown and reversed the EMT process. Overall, our results revealed that GTSE1 served as an oncogene regulating EMT through KLF4 in ccRCC, and that GTSE1 could also serve as a novel prognostic biomarker and may represent a promising therapeutic target for ccRCC.

High prevalence and reoccurrence rate of nephrolithiasis bring about serious socioeconomic and healthcare burden, necessitating the need of effective therapeutic agents. Previous study revealed that gallic acid (GAL) alters the nucleation pathway of calcium oxalate (CaOx). On the other hand, it appears protective role against oxidative injury. Whether GAL could protect against crystal-induced lesion in vivo, and its underlying mechanism is yet unsolved.This study aims to investigate the protective effects of GAL on the crystal-induced renal injury and its underlying mechanism in the mouse model of stone formation induced by glyoxylic acid.The mouse model of stone formation was established via successive intraperitoneal injection of glyoxylate. Proximal tubular epithelial cell line HK-2 treated with calcium oxalate monohydrate (COM) was used as in vitro model. The protective role of GAL on nephrolithiasis was tested by determination of tubular injury, crystal deposition and adhesion, levels of inflammatory cytokines, macrophage infiltration and the redox status of kidney. In vitro, effect of GAL on the ROS level and oxidative tubular injury induced by COM were detected, as well as major antioxidant pathway Nrf2/HO-1.Administration of GAL alleviates the renal deposition and adhesion of CaOx stone. Meanwhile, GAL ameliorates the inflammation and renal tubular injury. Level of intracellular ROS, osteopontin and CD44 are reduced, either in the mouse model of stone formation or in the COM-treated HK-2 cells after treatment of GAL. Mechanistically, GAL activates Nrf2/HO-1 pathway in HK-2 cells. Silencing Nrf2 abrogates the protective effect of GAL on the oxidative injury and adhesion of COM in HK-2 cells.Taken together, our study demonstrates the protective effect of GAL on the deposition of kidney stone and consequent tubular injury. Induction of the antioxidant pathway Nrf2/HO-1 was found to decrease the level of ROS and oxidative injury, thus implying that GAL could be a potential therapeutic agent for the treatment of nephrolithiasis.

Background OTUB1 (ovarian tumor domain protease domain-containing ubiquitin aldehyde-binding proteins)-mediated deubiquitination of FOXM1 (Forkhead box M1) participates in carcinogenesis of various tumors. We aim to investigate the effect and mechanism of OTUB1/FOXM1 on RCC (renal cell carcinoma) progression. Expression levels of OTUB1 in RCC tissues and cell lines were examined by qRT-PCR (quantitative real-time polymerase chain reaction) and immunohistochemistry. Cell proliferation was measured with CCK8 (Cell Counting Kit-8) and colony formation assays. Wound healing and transwell assays were used to determine cell migration and invasion, respectively. The effect of OTUB1 on FOXM1 ubiquitination was examined by Immunoprecipitation. Western blot was used to uncover the underlying mechanism. In vivo subcutaneous xenotransplanted tumor model combined with immunohistochemistry and western blot were used to examine the tumorigenic function of OTUB1. Results OTUB1 was up-regulated in RCC tissues and cell lines, and was associated with poor prognosis of RCC patients. Knockdown of OTUB1 inhibited cell viability and proliferation, as well as migration and invasion of RCC cells. Mechanistically, knockdown of OTUB1 down-regulated FOXM1 expression by promoting its ubiquitination. Down-regulation of FOXM1 inhibited ECT2 (epithelial cell transforming 2)-mediated Rho signaling. Moreover, the inhibition of RCC progression caused by OTUB1 knockdown was reversed by FOXM1 over-expression. In vivo subcutaneous xenotransplanted tumor model also revealed that knockdown of OTUB1 could suppress in vivo RCC growth via down-regulation of FOXM1-mediated ECT2 expression. Conclusions OTUB1-mediated deubiquitination of FOXM1 up-regulates ECT-2 to promote tumor progression in RCC, providing a new potential therapeutic target for RCC treatment.

Lactate dehydrogenase A (LDHA), a key enzyme regulating aerobic glycolysis, is overexpressed in many human cancers, and correlates with poor clinical outcomes. Aerobic glycolysis is a hallmark of cancer, and drugs targeting its regulators, including LDHA, are being developed. However, the mechanisms of LDHA inhibition and the physiological significance of the LDHA inhibitors in cancer cells are unclear. Here, we show that microRNA-30a-5p (miR-30a-5p) suppresses LDHA expression by directly targeting its 3'-UTR. Through inhibition of LDHA, miR-30a-5p dampens glycolysis by decreasing glucose uptake, lactate production, ATP generation, and extracellular acidification rate (ECAR), and increasing oxygen consumption rate (OCR) in breast cancer cells. Importantly, glycolysis regulated by miR-30a-5p is critical for its regulating breast tumor growth and metastasis both in vitro and in vivo. In breast cancer patients, miR-30a-5p expression is negatively correlated with LDHA expression. Moreover, patients who had increased glucose uptake in tumors assessed by PET scans showed decreased miR-30a-5p expression and increased expression of LDHA. Our findings provide clues regarding the role of miR-30a-5p as a tumor suppressor in breast cancer through the inhibition of LDHA. Targeting LDHA through miR-30a-5p could be a potential therapeutic strategy in breast cancer.

Previous studies have shown that the stroke volume variation (SVV), the pulse pressure variation (PPV) and the pleth variability index (PVI) could be successfully used for predicting fluid responsiveness (FR) in surgical patients. The aim of this study was to validate the ability of SVV, PPV and PVI to predict intraoperative FR in mechanically ventilated patients with obstructive jaundice (OJ). Thirty-two patients with OJ (mean serum total bilirubin 190.5 ? 95.3??mol L?1) received intraoperative volume expansion (VE) with 250?ml colloids immediately after an exploratory laparotomy had been completed and after a 5?min period of hemodynamic stability. Hemodynamic variables were recorded before and after VE. FR was defined as an increase in stroke volume index > 10% after VE. The ability of SVV, PPV and PVI to predict FR was assessed by calculation of the area under the receiver operating characteristic curve. Eleven (34%) patients were responders and 21 patients were nonresponders to VE. The PPV was the unique dynamic index that had the moderate ability to predict FR during surgical procedures, the area under the curve was 0.71 (95% CI, 0.523 to 0.856; P = 0.039) and the threshold (sensitivity and specificity) discriminated responders was 7.5% (63.6%/71.4%). The present study concluded that SVV and PVI were not reliable predictors of FR, but PPV has some value predicting FR in patients with OJ intraoperatively.

// Yuanyuan Zhang 1, * , Haixing Mai 2, * , Gang Guo 3 , Guofang Bi 1 , Guangtao Hao 1 , Yuanyuan Li 1 , Xiaofang Wang 1 , Longmei Cheng 1 , Jing Wang 1 , Ruihua Dong 1 , Zeyuan Liu 1 , Lijun Chen 2 and Hengyan Qu 1 1 Department of Clinical Pharmacology, Academy of Military Medical Sciences Affiliated Hospital, 307 Clinical College, Anhui Medical University, Beijing 100071, China 2 Department of Urology Department, Academy of Military Medical Sciences Affiliated Hospital, Beijing 100071, China 3 Department of Urology Department, The General Hospital of the People's Liberation Army, Beijing 100853, China * These authors contributed equally to this work Correspondence to: Hengyan Qu, email: quhengyan@hotmail.com Keywords: single-nucleotide polymorphisms; cell-free DNA; pharmacokinetics; sunitinib; renal-cell carcinoma Received: December 27, 2016 Accepted: November 11, 2017 Epub: January 03, 2018 Published: March 06, 2018 ABSTRACT Background: Sunitinib is a tyrosine kinase inhibitor with effective therapeutic outcomes in patients with renal-cell carcinoma. The study were to analyze the association of single-nucleotide polymorphisms present in cell-free DNA and pharmacokinetics with sunitinib treatment-emergent adverse events in Chinese patients with renal-cell carcinoma. Materials and Methods: We genotyped 8 keys SNPs in 6 candidate genes. The plasma concentrations of sunitinib and N-desethyl sunitinib were measured using a high performance liquid chromatography-tandam mass spectrometry method. Correlations between the single-nucleotide polymorphisms and adverse events were investigated by univariate and multivariate logistic regression and we quantitatively evaluated the effect of single-nucleotide polymorphisms on the pharmacokinetics of sunitinib by using a population PK model. Results: Necessary dose reductions of sunitinib were significantly correlated with SNP rs1933437 in FLT3 . A higher severity of AEs were collected with SNP rs2032582 in ABCB1 and rs1800812 in PDGFRα . Thrombocytopenia was collected with rs1800812 in PDGFRα . Our study provides a population PK model of sunitinib with the ABCB1 genotype as a predictive covariate for apparent oral clearance. Conclusions: Our study preliminarily confirmed the hypothesis that the pharmacokinetics of sunitinib is affected by the SNPs of enzyme in Chinese renal-cell carcinoma patients, and this affects the different distribution and severity of adverse events of sunitinib.

This single-center, observational study analyzed the association between plasma concentration of sorafenib and its safety and efficacy in Chinese patients with metastatic renal cell carcinoma (mRCC). Adult patients with RCC (n = 94), treated with sorafenib were enrolled between January 2014 and January 2015. Sorafenib plasma concentrations were measured by liquid chromatography-tandem mass spectrometry. Safety and efficacy variables were evaluated using National Cancer Institute-Common Toxicity Criteria for Adverse Events and Response Evaluation Criteria in Solid Tumors criteria. Association of plasma concentration with safety and efficacy was analyzed. The steady state plasma concentration of sorafenib after 2 weeks of treatment ranged from 881 to 12,526 ng/mL. Major adverse reactions (ADRs) included diarrhea (76.5%), hand-foot syndrome (HFS; 68.99%) and fatigue (55.32%). Significant association was reported between plasma concentration and all the ADRs except rash. At 6 weeks, complete response (CR), partial response (PR), stable disease (SD) and progressive disease (PD) was reported in 3.1%, 13.82%, 52.2% and 13.82% patients, respectively. Objective response and disease control rates were 17.02% and 69.14%. Plasma concentration of sorafenib was >10,000 ng/mL in patients with severe ADRs, which decreased with reduction in dose or discontinuation of treatment. After 21.2 weeks follow-up, median progression free survival was 12.3 months. CR, PR, SD and PD were reported in 1%, 46%, 33% and 19% patients. In conclusion, plasma concentration of sorafenib was associated with its safety and efficacy in Chinese patients with mRCC.

To investigate the effect of sunitinib on the expressions of co-stimulatory molecule ligands, programmed death ligand 1 (PD-L1), PD-L2, CD80, CD86, B7-H4 and herpes virus entry mediator (HVEM) on peripheral blood monocyte-derived dendritic cells (DCs) from patients with renal cell carcinoma (RCC).Monocyte-derived DCs from patients with RCC were cultured in vitro and randomly divided into three groups: sunitinib combined with lipopolysaccharide (LPS), LPS only and dimethyl sulfoxide (DMSO) treatment. Sunitinib plus LPS group was pretreated with 200 ng/mL sunitinib for 12 hours followed by stimulated with 1 μg/mL LPS for 24 hours; LPS group was pretreated with 1 μL/mL DMSO for 12 hours and then stimulated with 1 μg/mL LPS for 24 hours; DMSO group was treated with 1 μL/mL DMSO for 36 hours. Morphological changes were observed by an inverted microscope. Flow cytometry was used to detect the expressions of PD-L1, PD-L2, CD80, CD86, B7-H4 and HVEM.Sunitinib-LPS co-treated and LPS-treated cells had typical dendrites, while DMSO-treated cells had no obvious dendrites. Compared with the LPS-treated group, the expressions of CD80, PD-L1 and B7-H4 on DCs significantly decreased in the sunitinib-LPS group; the expressions of PD-L1, PD-L2, CD80, CD86, B7-H4 and HVEM were lower in the DMSO group.Sunitinib inhibits the expressions of CD80, PD-L1 and B7-H4 on DCs induced by LPS.

Summary Aerobic glycolysis (the Warburg effect) facilitates tumor growth, and drugs targeting aerobic glycolysis are being developed. However, how the Warburg effect is directly regulated is largely unknown. Here we show that transcription factor SIX1 directly increases the expression of many glycolytic genes, promoting the Warburg effect and tumor growth in vitro and in vivo . SIX1 regulates glycolysis through HBO1 and AIB1 histone acetyltransferases. Cancer-related SIX1 mutation increases its ability to promote aerobic glycolysis and tumor growth. SIX1 glycolytic function is directly repressed by microRNA-548a-3p, which is downregulated, inversely correlates with SIX1, and is a good predictor of prognosis in breast cancer patients. Thus, the microRNA-548a-3p/SIX1 axis strongly links aerobic glycolysis to carcinogenesis and may become a promising cancer therapeutic target.

To observe the changes of programmed death-1 ligand 1 (PD-L1) and PD-L2 expressions on mouse bone marrow-derived dendritic cells (DCs) stimulated by sunitinib.DCs were randomly divided into four groups which were treated with sunitinib (100, 200, 300 ng/mL) and dimethylsulfoxide (DMSO), respectively. After 48 hours, PD-L1 and PD-L2 expression levels were analyzed by flow cytometry.Compared with the control group, the expression of PD-L1 on mature DCs (mDCs) and all DCs [including mature DCs and immature DCs (imDCs)] was significantly down-regulated in sunitinib treatment groups. The PD-L1 expression percentages of imDCs, mDCs and DCs were significantly reduced in sunitinib treatment groups; the percentage of mDCs expressing PD-L2 also dropped in all treatment groups, and the percentage of DCs expressing PD-L2 decreased in 100 and 300 ng/mL sunitinib treatment groups.Sunitinib can significantly reduce the expressions of PD-L1 and PD-L2 on mouse DCs.

To investigate the relationship between the number and maturation of dendritic cell (DC) and microvessel density (MVD) in clear cell renal cell carcinoma (CCRCC).CCRCC paraffin-embedded tissues and surrounding normal tissues were collected from 30 cases of CCRCC who underwent operations in No. 307 hospital of PLA during July, 2010 to January, 2013. Immunohistochemistry was used to detect the expressions of CD11c, MHC-II, and CD34 to analyze the relationship among them.The positive expression rate of CD11c in CCRCC tissues [(93.08±66.14)%] was higher than that in the surrounding tissues [(25.91±12.29)%]. MVD in CCRCC tissues (26.31±11.05) was significantly higher than that in the surrounding tissues (12.78±5.49). The permillage of MHC-II positive DC in CCRCC tissues [(9.65±3.61)%] was significantly lower than that in the surrounding tissues [(17.60±6.26)%]. There was a negative correlation between the MVD value and the expression of MHC-II; there was no correlation between the MVD and the expression of CD11c. Positive expressions of various indicators in CCRCC tissues were not related to patients' age, sex, and TNM classification.In CCRCC tissues, there were higher MVD and more DCs than in the surrounding tissues, yet there were less mature DCs. There was a negative correlation between the MVD and the maturation of DC.

Hematopoietic pre-B cell leukemia transcription factor (PBX)-interacting protein (HPIP) was shown to be crucial during the development and progression of a variety of tumors. However, the role of HPIP in renal cell carcinoma (RCC) is unknown. Here we report that HPIP is upregulated in most RCC patients, positively correlates with tumor size, high Fuhrman grade and preoperative metastasis, and predicts poor clinical outcomes. Mechanistically, we identified casein kinase 1α (CK1α), a critical regulator of tumorigenesis and metastasis, as a novel HPIP-interacting protein. HPIP facilitates RCC cell growth, migration, invasion and epithelial–mesenchymal transition depending on its interaction with CK1α. Activation of mammalian target of rapamycin pathways by HPIP is partly dependent on CK1α and is required for HPIP modulation of RCC cell proliferation and migration. HPIP knockdown suppresses renal tumor growth and metastasis in nude mice through CK1α. Moreover, expression of CK1α is positively correlated with HPIP in RCC samples, and also predicts poor clinical outcome-like expression of HPIP. Taken together, our data demonstrate the critical regulatory role of the HPIP–CK1α interaction in RCC, and suggest that HPIP and CK1α may be potential targets for RCC therapy.