Study on the regulation of microRNA-375 on PDK1 expression in brain tissue of scrapie agent 139 A infected mice / 中华实验和临床病毒学杂志

Objective@#To investigate the regulation of microRNA-375(miR-375) on the expressions of 3’-phosphoinositide-dependent kinase 1 (PDK1) in the brain tissues of scrapie agent 139 A infected mice.@*Methods@#PDK1 protein in 139 A infected mice brain tissue was detected by WB and immunochemistry. The change of microRNA-375 was detected by reverse transcription polymerase chain reaction (RT-PCR) and next-generation sequencing method . The pmiR-REPORT reporter system was used to value the regulation of miR-375 on PDK1 3’-untranslated region (3’UTR).@*Results@#The expression of PDK1 in the brain tissue of 139 A infected mice was significantly increased as compared to that of control group, while the expression of miR-375 was decreased. The result of pmiR-REPORT reporter system showed that PDK1 3’UTR was the regulation target of miR-375.@*Conclusions@#The expression of PDK1 in the brain tissue of 139 A infected mice was significantly increased, which was probably related to the regulation of miR-375 on the 3’UTR of PDK1.

MicroRNA-375 Suppresses the Tumor Aggressive Phenotypes of Clear Cell Renal Cell Carcinomas through Regulating YWHAZ / 中华医学杂志(英文版)

Background@#MicroRNAs (miRNAs) are key regulators during tumor initiation and progression. MicroRNA-375 (MiR-375) has been proven to play a tumor-suppressive role in various types of human malignancies; however, its biological role in clear cell renal cell carcinoma (ccRCC) remains unclear. The purpose of this study was to explore the biologic role as well as the underlying mechanism of miR-375 in ccRCC progression.@*Methods@#Quantitative polymerase chain reaction (qPCR) was applied to test the expression of miR-375 in tissues and cell lines by t-test. Functional experiments were used to investigate the biological role of miR-375 utilizing a gain-of-function strategy. The target of miR-375 was investigated by bioinformatic analysis and further verified by luciferase reporter assay, qPCR, Western blotting, and functional experiments in vitro.@*Results@#Our study demonstrated that miR-375 was significantly downregulated in ccRCC tissues (cancer vs. normal, 0.804 ± 0.079 vs. 1.784 ± 0.200, t = 5.531 P < 0.0001) and cell lines, and loss of miR-375 expression significantly associated with advanced Fuhrman nuclear grades (Grade III and IV vs. Grade I and II, 1.000 ± 0.099 vs. 1.731 ± 0.189, t = 3.262 P = 0.003). Functional studies demonstrated that miR-375 suppressed ccRCC cell proliferation, migration, and invasion (all P < 0.05 in both 786-O and A498 cell lines). Multiple miRNA target prediction algorithms indicated the well-studied oncogene YWHAZ as a direct target of miR-375, which was further confirmed by the luciferase reporter assay, qPCR, and Western blotting. Moreover, restoration of YWHAZ could rescue the antiproliferation effect of miR-375.@*Conclusions@#The data provide the solid evidence that miR-375 plays a tumor-suppressive role in ccRCC progression, partially through regulating YWHAZ. This study expands the antitumor profile of miR-375, and supports its role as a potential therapeutic target in ccRCC treatment.

miR-375 promotes prostate cancer cell migration and invasion by targeting KLF4 / 重庆医学

Objective To explore the expression,role and mechanism of miR-375 in prostate cancer (PCa) cells.Methods PCa cells were cultured and transfected with plasmid,the migration and invasion of PCa were detected by Transwell;the expression of miR-375 and KLF4 mRNA were detected by qPCR;the expression of KLF4 were detect by Western blot;the potential target genes of miR-375 were analyzed by bioinformatics and verified by dual luciferase report.Results The expression of miR-375 were significantly up-regulated in the PCa;Inhibited the expression of miR-375 could significantly inhibit the migration and invasion of PCa cells.KLF4 was the potential target genes of miR-375,which verified through bioinformatics analysis and dual luciferase report.The expression of KLF4 were significantly down-regulated in the PCa.Inhibited the expression of miR-375 could significantly up-regulated the expression of KLF4.Inhibited the KLF4 expression was able to reverse the suppressive effect miR-375 has on the migration and invasion of PCa cells.Conclusion miR-375 promotes the migration and invasion of PCa via inhibiting the expression of KLF4 and play the oncogene role.MiR-375 can be used as therapeutic targets for PCa,and provide a new direction for the treatment of PCa.

Analysis of gene network regulated by microRNA-375 in HCC / 中国病理生理杂志

AIM: To investigate the expression of microRNA-375 (miR-375) in hepatocellular carcinoma (HCC) and to analyze the target genes and signaling pathways regulated by miR-375.METHODS: The expression of miR-375 was examined at tissue microarray of HCC by in situ hybridization.The whole human genome chip and bioinforma-tics analysis were applied to screen out the differential expression genes and signaling pathways in 4 HCC cell lines trans-fected with miR-375 mimic.RESULTS:In situ hybridization showed the expression of miR-375 in HCC tissues were obvi-ously higher than that in tumor-adjacent tissues (P<0.05).There were 20 co-upregulated genes and 17 co-downregulated genes in all 4 cell lines.Bioinformatic analysis showed that there were 54 signaling pathways related to up-regulated genes and 48 signaling pathways related to down-regulated genes in all 4 cell lines.CONCLUSION: miR-375 may play a key role in the pathological process of HCC.The bioinformatic analysis is able to screen the target genes and signaling pathways regulated by miR-375 and to provide an explicit direction for further mechanism research on HCC.

Anti-apoptosis effect of liraglutide on islet via regulating microRNA-375 / 中国病理生理杂志

AIM: To observe the anti-apoptosis effect of liraglutide on the islet through microRNA-375 (miR-375) for providing additional pharmacodynamic evidence for its clinical application.METHODS: For in vivo study, C57BL/KsJ-db/m mice aged 8 weeks served as normal control group.A total of 40 male genetically diabetic C57BL/KsJ-db/db mice at the same age were randomly divided into diabetic control group (the db/db mice were injected subcutaneous-ly with equivalent amount of saline) and liraglutide group (the db/db mice were injected subcutaneously with liraglutide at dose of 300 μg? kg-1? d-1 ).After 8 weeks of administration, body weight (BW) was measured and blood was collected for detection of fasting blood glucose (FBG), fasting blood insulin (FINS), triglyceride (TG), total cholesterol (TC) and low-density lipoprotein cholesterol (LDL-C).Before sacrifice, intraperitoneal glucose tolerance test (IPGTT) and insulin tolerance test (ITT) were conducted.The histopathological features in the islet tissue were examined with HE staining.The apoptosis in the islet tissue was detected by TUNEL staining.The protein levels of caspase-3, Bcl-2 and Bax were deter-mined by Western blot.The level of miR-375 in the islet tissue was detected by qPCR.For in vitro study, the MIN-6 cells were cultured and divided into control group (incubated with equivalent amount of solvent), miR-375 mimic group and miR-375 mimic +liraglutide group.The cell viability was examined by MTT assay.The protein levels of caspase-3, Bcl-2 and Bax were detected by Western blot.RESULTS: In the in vivo study, compared with control group, the levels of BW, FBG, FINS, TC, TG and LDL-C were decreased significantly in liraglutide group.The islet apoptosis was reduced by the administration of liraglutide.The expression of Bcl-2 was up-regulated significantly, while the protein levels of caspase-3 and Bax were down-regulated significantly in liraglutide group.The level of miR-375 was decreased significantly.In the in vitro study, the cell viability was decreased in miR-375 mimic group and increased in miR-375 mimic +liraglutide group. Moreover, the expression of Bcl-2 was decreased and the protein levels of caspase-3 and Bax were increased with the incu-bation of miR-375 mimic, while the expression of Bcl-2 was increased and the protein levels of caspase-3 and Bax were de-creased with the co-incubation of miR-375 mimic and liraglutide.CONCLUSION: Liraglutide attenuates islet apotosis, and the mechanism may be associated with its effects of reducing the elevated level of miR-375 in islet tissues.

Effect of miR-375 on viability, cell cycle and apoptosis of colon cancer HCT116 cells / 中国病理生理杂志

[ ABSTRACT] AIM:To investigate the effect of microRNA-375 ( miR-375) on the viability, cell cycle and apop-tosis of HCT116 cells.METHODS: The expression of miR-375 in different colorectal cancer cell lines was detected by real-time PCR.The miR-375 mimics was transfected into HCT116 cells by LipofectamineTM 2000.The mRNA expression of miR-375 and AEG-1 was detected by real-time PCR.The HCT116 cell viability was detected by MTT assay.The changes of apoptosis and cell cycle distribution were analyzed by flow cytometry.RESULTS:Real-time PCR showed that miR-375 expression was the lowest in HCT116 among 4 colorectal cancer cell lines.The expression level of miR-375 significantly in-creased in miR-375 mimics group compared with that in the negative control group.The high expression level of miR-375 significantly inhibited the mRNA expression of AEG-1.After transfection with miR-375 mimics, the cell viability was in-hibited, the apoptotic rate was increased, the proportion of G1-stage cells was increased, and the proportion of S-stage cells was decreased.CONCLUSION:miR-375 inhibits the viability, mediates the cell cycle arrest and promotes the apoptosis of colon cancer HCT116 cells.miR-375 may act as a tumor suppressor in colorectal cancer by inhibiting AEG-1.