Association of RGS20 expression with the progression and prognosis of renal cell carcinoma.

Regulator of G protein signaling 20 (RGS20) has been shown to be highly expressed in various types of cancer. The present study aimed to investigate the effects of RGS20 in patients with renal cell carcinoma (RCC) and in RCC cells. Bioinformatics analysis was performed to analyze the role of RGS20 in RCC. Quantitative PCR and western blotting were used to determine the mRNA and protein expression levels of RGS20 in cells, respectively. After RGS20 inhibition, the proliferation, apoptosis, migration and invasiveness of A-498 cells were tested using MTT assay, EdU assay, propidium iodide staining, Annexin V-FITC/PI kit, wound healing assay and Transwell assay. High RGS20 expression was closely associated with the progression and immune infiltration of RCC, and may be considered as an independent indicator of poor prognosis in RCC. After knocking down RGS20, the proliferation, migration and invasiveness of cells were impaired, the cell cycle was arrested at the G0/G1 phase, and the level of apoptosis was increased. In addition, the mRNA expression levels of securin, CDC20 and cyclin B1 were decreased in RGS20-knockdown cells. RGS20 expression was significantly associated with the infiltration level of activated CD4 T cells, type 1 T helper cells and activated dendritic cells. In summary, RGS20 expression was associated with RCC progression and poor prognosis; thus, it may be used to estimate the prognosis of RCC and may serve as a new potential treatment strategy for RCC.

PTEN status determines chemosensitivity to proteasome inhibition in cholangiocarcinoma.

Patient-derived xenografts (PDXs) and PDX-derived cells (PDCs) are useful in preclinical research. We performed a drug screening assay using PDCs and identified proteasome inhibitors as promising drugs for cholangiocarcinoma (CCA) treatment. Furthermore, we determined that phosphate and tensin homology deleted on chromosome ten (PTEN) deficiency promotes protein synthesis and proteasome subunit expression and proteolytic activity, creating a dependency on the proteasome for cancer cell growth and survival. Thus, targeting the proteasome machinery with the inhibitor bortezomib inhibited the proliferation and survival of CCA cells lacking functional PTEN. Therapeutic evaluation of PDXs, autochthonous mouse models, and patients confirmed this dependency on the proteasome. Mechanistically, we found that PTEN promoted the nuclear translocation of FOXO1, resulting in the increased expression of BACH1 and MAFF BACH1 and MAFF are transcriptional regulators that recognize the antioxidant response element, which is present in genes encoding proteasome subunits. PTEN induced the accumulation and nuclear translocation of these proteins, which directly repressed the transcription of genes encoding proteasome subunits. We revealed that the PTEN-proteasome axis is a potential target for therapy in PTEN-deficient CCA and other PTEN-deficient cancers.

miR-489-3p Inhibits Prostate Cancer Progression by Targeting DLX1.

PURPOSE: Prostate cancer (PCa) is the third most common cancer in men and the second leading cause of cancer-related death in men. DLX1 belongs to the DLX homeobox family and exhibits antitumor activity in many kinds of tumors. MicroRNAs (miRNAs) play important roles in the progression of cancer. However, whether miRNAs affect the development of PCa by targeting DLX1 has not been determined. In this study, we aimed to investigate the role of miR-489-3p in the regulation of DLX1 expression and PCa progression and to provide a potential therapeutic target for PCa treatment. METHODS AND MATERIALS: The Cancer Genome Atlas database was used to analyze the divergent expression of DLX1 in carcinomas and adjacent normal tissues. The expression level of DLX1 in malignant and normal prostate cells was also measured using RT-qPCR and Western blotting. A dual-luciferase reporter assay was performed to determine whether miR-489-3p directly targets DLX1. We transfected 22Rv1 and DU145 cells with miR-489-3p mimics to overexpress miR-489-3p and then evaluated its effect on cellular function. MTT, EdU, colony formation and cell cycle assays were used to evaluate cell growth. JC-1 and ROS assays with flow cytometry were performed to indirectly analyze apoptosis. Transwell assays were conducted to investigate metastasis. RESULTS: The expression level of DLX1 was upregulated in both PCa tissues and cell lines. MiR-489-3p directly targeted DLX1 and downregulated its expression. Overexpression of miR-489-3p significantly suppressed cell growth. MiR-489-3p induced apoptosis through mitochondrial function impairment. Overexpression of miR-489-3p also inhibited cell migration and invasion. DLX1 overexpression reversed the above effects induced by miR-489-3p. CONCLUSION: We identified the involvement of the miR-489-3p/DLX1 pathway in PCa for the first time. In this pathway, miR-489-3p acts as a tumor suppressor by negatively regulating the expression of DLX1. MiR-489-3p may be a potential therapeutic target for PCa treatment.

RPB5-Mediating Protein Promotes Cholangiocarcinoma Tumorigenesis and Drug Resistance by Competing With NRF2 for KEAP1 Binding.

BACKGROUND AND AIMS: Cancer cell survival depends on the balance between reactive oxygen species production and scavenging, which is regulated primarily by NRF2 during tumorigenesis. Here, we demonstrate that deletion of RBP5-mediating protein (RMP) in an autonomous mouse model of intrahepatic cholangiocarcinoma (ICC) delays tumor progression. APPROACH AND RESULTS: RMP-overexpressing tumor cells exhibited enhanced tolerance to oxidative stress and apoptosis. Mechanistically, RMP competes with NRF2 for binding to the Kelch domain of KEAP1 (Kelch-like ECH-associated protein 1) through the E**E motif, leading to decreased NRF2 degradation via ubiquitination, thus increasing NRF2 nuclear translocation and downstream transactivation of antioxidant genes. This RMP-KEAP1-NRF2 axis promotes ICC tumorigenesis, metastasis, and drug resistance. Consistent with these findings, the RMP level in human ICC is positively correlated with the protein level of NRF2 and is associated with poor prognosis. CONCLUSION: These findings reveal that RMP is involved in the oxidative stress defense program and could be exploited for targeted cancer therapies.

Six2 is negatively correlated with prognosis and facilitates epithelial-mesenchymal transition via TGF-ß/Smad signal pathway in hepatocellular carcinoma.

BACKGROUND: Increasing evidence indicates that Six2 contributes to tumorigenesis in various tumor including hepatocellular carcinoma (HCC). This study aimed to determine the role of Six2 in HCC and to elucidate the association of Six2 with clinical pathological characteristics. METHODS: The expressions of Six2 in HCC tumor, para-tumor tissue and portal vein tumor thrombus (PVTT) were detected by tissue microarray technique, immunohistochemistry, real-time RT-PCR and Western blotting. Chi-square and Kaplan-Meier analysis were used to analyze the correlation between Six2 expression and prognosis of HCC patients. Lentivirus mediated Six2 knockdown, spheroid formation assay, proliferation assay and subcutaneous tumor implantation were performed to determine the function of Six2. RESULTS: In 274 HCC samples, Six2 was strongly expressed. Kaplan-Meier analysis revealed that high expression of Six2 was correlated with a shorter overall survival (OS) and disease-free survival (DFS). Moreover, Six2 expression was associated with sex, alpha-fetoprotein, tumor size and portal vein invasion. Six2 was highly expressed in PVTT. Six2 knockdown inhibited HCC cell lines proliferation, migration, and self-renewal in vitro and in vivo. In addition, low-expression of Six2 weakened TGF-ß induced Smad4 activation and epithelial-mesenchymal transition in HCC cell lines. CONCLUSIONS: Elevated Six2 expression in HCC tumor patients was associated with negative prognosis. Upregulated Six2 promoted tumor growth and facilitated HCC metastasis via TGF-ß/Smad signal pathway.

Identification of METTL14 in Kidney Renal Clear Cell Carcinoma Using Bioinformatics Analysis.

The kidney renal clear cell carcinoma (KIRC) with poor prognosis is the main histological subtype of the renal cell carcinoma, accounting for 80-90% of patients. Currently, the N6-methyladenosine (m6A) epitranscriptional modification draws much attention. The m6A RNA modification, the most plentiful internal modification of mRNAs and noncoding RNAs in the majority of eukaryotes, regulates mRNAs at different levels and is involved in disease occurrence and progression. The GTExPortal and TCGAportal were applied to investigate the METTL14 mRNA expression in different tissues and KIRC stages. The Human Protein Atlas was used to verify the location of METTL14 in KIRC tissues. The main microRNAs (miRNAs) related to KIRC were analyzed using OncoLnc and starBase, while corresponding circular RNAs (circRNAs) interacting with miRNAs were predicted via circBank; then, the METTL14-miRNA-circRNA interaction network was established. The level of methyltransferase-like 14 (METTL14) mRNA was significantly lower in KIRC tissues compared with normal kidney tissues, which was relative to clinical and pathological stages. circRNAs may regulate METTL14 mRNA as miRNAs sponge to affect the KIRC progression. METTL14 mRNA is likely to regulate PTEN mRNA expression via changing its m6A RNA modification level. METTL14 mRNA expression negatively correlated with the KIRC stages and positively correlated with KIRC patients' overall survival, which has great potential to serve as a clinical biomarker in KIRC.

Combination of Kras activation and PTEN deletion contributes to murine hepatopancreatic ductal malignancy.

Kras mutations are among the most common genetic abnormalities in human neoplasms, including cholangiocarcinomas, pancreatic cancer and colon cancer. PTEN has previously been associated with cholangiocarcinoma development in murine models. Here, we have established novel mouse models of neoplasms by liver-specific and biliary-pancreatic Kras activation and PTEN deletion. By liver-specific disruption of PTEN and activation of Kras in mice caused rapid development of intrahepatic biliary epithelial proliferative lesions (Intrahepatic cholangiocarcinoma, ICC), which progress through dysplasia to invasive carcinoma. In contrast, Kras activation in combination with heterozygous PTEN deletion induced mixed carcinomas of liver (both ICC and hepatocellular carcinoma, HCC), whereas Kras activation alone did not induce biliary tract neoplasm. Use of Sox9-Cre-LoxP-based approach to coordinately delete PTEN and activate Kras in the adult mouse resulted in not only development of low-grade biliary lesions (ICC and extrahepatic bile duct carcinoma, ECC) but also pancreatic carcinomas. Our data provide a functional link between PTEN gene status, hepatobiliary cell fate, and HCC, biliary carcinoma, pancreatic cancer pathogenesis, and present novel genetically engineered mouse models of PTEN loss-driven malignancy.

Clinical efficacy of digital flexible reteroscope and modular flexible ureteroscope combined with holmium laser lithotripsy for subrenal calyx calculus / 中国内镜杂志

Objective To evaluate the efficacy of flexible ureteroscopy combined with holmium laser lithotripsy for the treatment of subrenal calyx calculus, and comparing with digital flexible ureteroscope and modular flexible ureteroscope with holmium lithotripsy for the difference of the efficacy of the treatment of subrenal calyx calculus. Methods Review of the clinical data of 93 patients with subrenal calyx calculus, of which 48 cases were treated with digital flexible ureteroscope (digital flexible ureteroscope group, DFU group), 45 cases with modular flexible ureteroscope treatment (modular flexible ureteroscope group, MFU group), lithotripsy effect and complications were compared between the two groups. Results The mean operation time, one-session stone-free rate were significantly different between the two groups (P < 0.05). No significant differences in hospital stay, the success rate of looking for calculus, complication and hospitalization expense (P > 0.05). Conclusion With digital flexible ureteroscope and modular flexible ureteroscope treatment of subrenal calyx calculus all are safe and effective. The use of DFU than the use of MFU in the treatment of subrenal calyx calculus operation time is shorter, stone-free rateis higher, the effect is better. There is little difference between their hospitalization expenses, but the use of MFU can reduce the cost of the department.

Clinical efficacy of digital flexible reteroscope and modular flexible ureteroscope combined with holmium laser lithotripsy for subrenal calyx calculus / 中国内镜杂志

Objective To evaluate the efficacy of flexible ureteroscopy combined with holmium laser lithotripsy for the treatment of subrenal calyx calculus, and comparing with digital flexible ureteroscope and modular flexible ureteroscope with holmium lithotripsy for the difference of the efficacy of the treatment of subrenal calyx calculus. Methods Review of the clinical data of 93 patients with subrenal calyx calculus, of which 48 cases were treated with digital flexible ureteroscope (digital flexible ureteroscope group, DFU group), 45 cases with modular flexible ureteroscope treatment (modular flexible ureteroscope group, MFU group), lithotripsy effect and complications were compared between the two groups. Results The mean operation time, one-session stone-free rate were significantly different between the two groups (P < 0.05). No significant differences in hospital stay, the success rate of looking for calculus, complication and hospitalization expense (P > 0.05). Conclusion With digital flexible ureteroscope and modular flexible ureteroscope treatment of subrenal calyx calculus all are safe and effective. The use of DFU than the use of MFU in the treatment of subrenal calyx calculus operation time is shorter, stone-free rateis higher, the effect is better. There is little difference between their hospitalization expenses, but the use of MFU can reduce the cost of the department.

Elevated ZC3H15 increases HCC growth and predicts poor survival after surgical resection.

Zinc finger CCCH-type containing 15 (ZC3H15), also known as DRG family regulatory protein 1 (DFRP1), is a highly conserved eukaryotic protein that associates with active translation machinery. The aim of our study was to explore the clinical relevance and intrinsic functions of ZC3H15 in hepatocellular carcinoma (HCC). We constructed a cohort with 261 tumor and matched normal tissues from HCC patients. ZC3H15 protein and mRNA levels were determined using immunohistochemistry, western blot analysis, and quantitative polymerase chain reaction. ZC3H15 was highly expressed in the majority of HCC cases, and high ZC3H15 levels were significantly associated with high serum a-fetoprotein (AFP) levels (>20 ng/mL) and vascular invasion. Kaplan-Meier and Cox regression data indicated that elevated ZC3H15 was an independent predictor for HCC-specific disease-free survival (hazards ratio [HR], 1.789; 95% confidence interval [95% CI], 1.298-2.466 [P=0.0004]) and overall survival (HR, 1.613; 95% CI, 1.120-2.322 [P=0.0101]). Interaction of ZC3H15 with TRAF2 increased activation of NFκB signaling. These results suggest ZC3H15 is an independent prognostic marker in HCC patients that is clinicopathologically associated with tumor invasion and serum AFP levels.