| Objective: Renal Cell Carcinoma(RCC)is one of the most common malignant tumors in the urinary system,accounting for about 2-3% of all adult neoplasms.In recent years,studies have shown that miRNAs play a regulatory role in the development and progression of tumors.Some researches have revealed that miR-19b-1 is abnormally expressed in many malignant cancers.However,the functions of miR-19b-1 in RCC oncogene remain unknown at present.The aim of this experiment was to explore the expression and function of miR-19b-1 in RCC and to analyze the possibility of using miR-19b-1 as a novel biomarker of RCC.Methods: The 32 RCC tissues,paired normal tissues,RCC cell lines(786-O,ACHN)and normal renal cell line(293T)was chosen as specimen,from which total RNA was extracted.And then,the total RNA was reversed,and the expression of miR-19b-1 was revealed by RT-qPCR.The difference of miR-19b-1 expressions between tissues and RCC cell lines was analyzed by using statistical software.Patients were divided into high and low expression groups according to the mean expression of miR-19b-1,and the Fisher exact test was used to analyze the correlation of the expression level of miR-19b-1 between the age,sex,histological type and clinical stage of renal cell carcinoma.Expression of miR-19b-1 was up-regulated or down-regulated with transfecting miR-19b-1 mimic or inhibitor,respectively.The effects of miR-19b-1 on renal cell carcinoma cells proliferation,migration,invasion and apoptosis were detected by cell scratch assay,cell proliferation assay(MTT and CCK-8),transwell migration and invasion assay,flow cytometry and Hoechst33342 staining.Results: The consequences of RT-qPCR showed that miR-19b-1 expression in RCC tissues and RCC cell lines was obviously higher than in normal adjacent tissues and normal renal cell lines,respectively(**p<0.05).The expression level of miR-19b-1 was not related to age,sex and the type of renal cell carcinoma,except the clinical stage of patients(p<0.05).Expression of miR-19b-1 could be up-regulated or down-regulated when transfected with miR-19b-1 mimic or inhibitor.The validation of cell line transfection efficiency showed that the 786-O cell line was negative and the ACHN cell line was positive.Results of cell scratch assay and transwell migration and invasion assay revelaed that up-regulation of miR-19b-1 could promote the mobility of renal cell carcinoma cells and down-regulation suppressed.In the 786-O cell line,over-expression(mimic)or down-regulation(inhibitor)of miR-19b-1 had no obvious influence on cell apoptosis and the apoptotic rate had no significant differences.However,the apoptotic rate of the ACHN cell line when transfected with the miR-19b-1 inhibitor or inhibitor NC was significantly interfered and the rate was found to be 44.05% vs.37.87%(p < 0.05).Also,the apoptotic rate of the cell line transfected with the miR-19b-1 mimic or mimic NC was 30.54 vs.40.03(p < 0.05).Conclusion: When compared with the adjacent normal tissues,the expression level of miR-19b-1 in renal cell carcinoma was significantly higher,and the expression in renal cell carcinoma cell lines(786-O /ACHN)was also significantly higher than that in normal renal cell line(293T).what is more,the expression level of miR-19b-1 was related to the stage of renal cell carcinoma.The expression of miR-19b-1 was higher in patients with higher clinical stage.with higher clinical stage.Down-regulation of miR-19b-1 could significantly inhibit renal cell carcinoma cells(786-O)migration and invasion,but they have no influences on the apoptosis.Up-regulation of miR-19b-1 could promote the migration and invasion of renal cell carcinoma cells(ACHN)and has some effect on the apoptosis.The results of this experiment show that miR-19b-1,abnormally expressed in renal cell carcinoma and RCC cells,could regulate the biological process of renal cancer cells,which is related to the clinical stage of renal cancer.Therefore,miR-19b-1 played an important role in the development of renal cell carcinoma and might be used as a biomarker of renal cell carcinoma for clinical application. |
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