Effects And Mechanisms Of Hsa-miR-19b-1-5p In Human Prostate Cancer

Objectives:Prostate cancer is one of the most common malignancies in middle-aged men.A few years ago,the incidence of prostate cancer in China was still at a very low level,but with the gradual improvement of people's living standards and the continuous improvement of the level of diagnosis,the incidence of prostate cancer and detective rate began to increase year by year,and it has become an important disease threatening the health of middle-aged men.This study was designed to investigate the effect of miRNA-19b-1-5p on the biological behavior of human prostate cancer cells,and to elucidate the effects and mechanisms of miRNA-19b-1-5p in human prostate cancer.Methods:Clinical specimens were collected and divided into prostate cancer group and benign prostatic tissue group.The specimens were 77 cases and 68 cases.Real-time PCR was used to detect the expression of miRNA-19b-1-5p in each tissue,and to analyze the correlation of the relative expression of miRNA-19b-1-5p with Gleason score and clinicopathological feature.The mimic and inhibitor of mi RNA-19b-1-5p were transfected into prostate cancer cell lines respectively by chemical transfection reagent.The Cell Counting Kit-8 was used to detect the proliferation ability of the cells.Transwell chamber was used to detect the invasion and metastasis abilities of the cells.The cell migration ability was measured by wound healing experiment and the cell apoptosis was analyzed by flow cytometry.The potential target genes of miRNA-19b-1-5p were predicted by using target gene prediction software such as miRanda,TargetScan and so on.Real-time PCR was used to detect the relative expression of miRNA-19b-1-5p mRNA and target gene mRNA in the cell lines after transfected by the mimic or inhibitor.And to analyze the relationship between them.Western blot was used to detect the change of the target gene.Results:MiRNA-19b-1-5p was highly expressed in benign prostatic tissues and was lower in prostate cancer tissues.There was significant difference between the two groups.The relative expression of miRNA-19b-1-5p was correlated with tumor Gleason score,clinicopathological feature.CCK-8 assay showed that the cell proliferation rate transfected by miRNA-19b-1-5p-mimic was significantly lower than that of the miRNA-19b-1-5p-inhibitor.In the Transwell assay,the number of cells transfected by miRNA-19b-1-5p-mimic was significantly less than that of the mi RNA-19b-1-5p-inhibitor group.In the wound healing test,the cell closure rate transfected by miRNA-19b-1-5p-mimic group was significantly smaller than that of the miRNA-19b-1-5p-inhibitor group.Flow cytometry showed an increase in the apoptotic rate transfected by miRNA-19b-1-5p-mimic group.The target gene software predicts that CXCR6 is a downstream target gene for miRNA-19b-1-5p,the results of Real-time PCR showed that miRNA-19b-1-5p was negatively correlated with CXCR6.Western blot also confirmed that CXCR6 expression was inhibited transfected by miRNA-19b-1-5p-mimic.Conclusions:MiRNA-19b-1-5p is low in prostate cancer and plays a role as the tumor suppressor gene.With the increase of the Gleason score and the level of clinicopathologicalfeature,the expression of miRNA-19b-1-5p was decreased.Upregulation of miRNA-19b-1-5p significantly inhibited the proliferation,invasion and metastasis of prostate cancer cell lines,and promoted the apoptosis of prostate cancer cells,which may be achieved by inhibiting the expression of downstream target gene CXCR6.