The Function And Mechanism Of MiRNA-587 In Hepatocellular Carcinoma

AimIn this study,we aim to study the expression level of miRNA-587 in HCC and its relationship with clinical features and ultrasound imaging characteristics;to study the biological function of miRNA-587 in HCC hepatocellular carcinoma cells;to study the targeting gene of miRNA-587 in HCC SMMC-7721 cells;to study the expression level of targeting gene RPSA in HCC and its relationship with clinical features and ultrasound imaging characteristics;to evaluate the relationship between RPSA expression level and all the characteristics with prognosis of HCC.MethodsPart One1 Utilizing bioinformatics and PCR assay about the clinical collection of 86 pairs of HCC cancer tissues and adjacent tissues to verify the expression level of microRNA-587 in HCC;2 Using the chi-square test to study the relationship between the expression of miRNA-587 and the clinical ultrasonographic features of 86 patients with HCC.Part Two1 The SMMC-7721 cells were transfected with the transfection reagents mi-587 mimic,mi-587 inhibitor,mimic negative control and inhibitor negative control.The proliferation,apoptosis and invasion function of SMMC-7721 cells were observed by MTT cell proliferation test,scratch test,clone formation test,transwell cell invasion test and flow cytometry.Part Three1 Bioinformatics predicts the targeting effect of microRNA-587 in HCC and using double luciferase assay to verify it;2 Both bioinformatics and PCR assay about the clinical collection of 86 pairs of cancer tissues of HCC and adjacent tissues were used to verify the expression level of the target gene RPSA about miRNA-587;3 The relationship between RPSA expression level and clinical ultrasonographic features of 86 patients with HCC was studied by chi-square test;4 Survival curves were drawn by the Kaplan-Meier method.Log Rank statistical method was used to explore the difference of survival time between HCC patients with RPSA high expression group and low expression group;5 The relationship between the expression of RPSA,the clinical features,the ultrasonographic features and the prognosis of HCC was studied by Cox proportional hazard regression model using univariate analyses and multivariate analyses.ResultsPart One1 Bioinformatics database GEO showed that the expression of miRNA-587 in HCC tissues was lower than that in non-tumor tissues(P<0.05),and the microRNA-587 gene was confirmed as an anti-oncogene of HCC in 86 HCCpatients’ cancer tissues and adjacent tissues by PCR analysis,which was consistent with the results of bioinformatics(P<0.05);2 In the relationship between miRNA-587 and clinical ultrasonographic features of HCC patients,microRNA-587 was only associated with AFP >400ng/ml(P<0.05).Part Two1 MiRNA-587 gene inhibits the proliferation,migration and invasion of SMMC-7721 cells(P<0.05),retards the cell cycle of hepatocellular carcinoma at G0/G1 phase,inhibits the proliferation of SMMC-7721 cells to S phase(P<0.05),and has no effect on the cloning and apoptosis of hepatocellular carcinoma cells(P>0.05).Part Three1 Bioinformatics analysis and double luciferase assay concluded that microRNA-587 targeted the regulation of RPSA gene expression;2 Bioinformatics analysis and PCR assay of 86 paired tissues showed that RPSA was more expressed in hepatocellular carcinoma than in adjacent tissues(P<0.05);3 In a series of clinical features and ultrasound imaging features of HCC patients,the expression of RPSA gene was only significantly different to alcohol status(P<0.05).Drinking alcohol may be a contributing factor to the high expression of RPSA;4 The level of RPSA gene is related to the survival of patients with HCC(P<0.05),the total survival time of the high expression group is shorter than that of the low expression group;5 Univariate analysis showed that among the clinical features of HCC patients,BCLC stage and TNM stage were the risk factors affecting theprognosis(P<0.05).And among the features of ultrasonography,the risk factors affecting the prognosis of HCC included the maximum diameter of tumors,the number of tumors and the combination of cancer thrombus(P<0.05).Further multivariate analysis showed that the number of tumors and the presence of tumor thrombus were independent risk factors for the prognosis of HCC(P<0.05).ConclusionThe expression of miRNA-587 is depressed in HCC,and it can inhibit the growth,migration and invasion of hepatocellular carcinoma cells by directly targeting RPSA.Moreover,the carcinogenesis of RPSA is related to the prognosis of HCC patients.The clinical BCLC staging,TNM staging and the maximum diameter measured by ultrasound,the number of tumors,and the combination of tumor thrombus are the prognostic risk factors of HCC.Tumor thrombus and the number of tumors are independent risk factors affecting the prognosis of HCC.