Effect of miR-1303 on the proliferation and migration of renal carcinoma 786-O cells through targeted inhibition of LPAR3 and its mechanism / 国际外科学杂志

ABSTRACT

Objective:To explore the mechanism by which microRNA (miRNA) -1303 inhibits the proliferation and migration of renal cell carcinoma 786-O cells through targeted regulation of lysophosphatidic acid receptor 3 (LPAR3) expression.Methods:quantitative real-time polymerase chain reaction (qRT-PCR) was used to detect the relative expression of miR-1303 in renal cancer cell lines (A498, ACHN, 786-O, OS-RC-2) and normal renal tubular epithelial cells HK-2. The miR-1303 mimic and the negative control sequence were transfected into the renal cancer cells with the lowest expression of miR-1303, respectively, as the miR-1303 group and the negative control group. qRT-PCR detected the relative expression of miR-1303 in the two groups of cells. MTT method and Transwell migration experiment were used to detect cell proliferation and migration ability. RegRNA 2.0 predicted the target genes of miR-1303. The dual luciferase reporter gene detected the binding of miR-1303 to the target gene. qRT-PCR and Western blotting detected the relative expression of LPAR3. Measurement data were expressed as mean±standard deviation ( ± s), t-test was used for comparison between two groups, and one-way analysis of variance was used for comparison between multiple groups. Results:The expressions of miR-1303 in renal cancer cell lines A498, ACHN, 786-O, OS-RC-2 and normal renal tubular epithelial cells HK-2 were 0.51±0.04, 0.79±0.02, 0.21±0.04, 0.55±0.07 and 1.00±0.05, the expression of miR-1303 in renal cancer cell lines was lower than that in HK-2 ( P<0.05), and the relative expression in 786-O cells was the lowest ( F=29.50, P<0.01). Compared with the control group, the expression of miR-1303 in the experimental group was significantly increased [(1.00±0.01) vs (7.98±0.88), t=7.95, P<0.01]. The cell absorbance value of the experimental group was significantly lower than that of the control group ( P<0.05). The number of cell migration in the experimental group was significantly lower than that in the control group ( P<0.05). miR-1303 can bind to LPAR3 mRNA in a complementary pair ( P<0.01). Compared with the control group, the expression of LPAR3 mRNA in the 786-O cells of the experimental group was significantly reduced [(1.00±0.01) vs (0.23±0.03), t=23.56, P<0.01]. Conclusion:miR-1303 may inhibit the proliferation and migration ability of renal cancer 786-O cells by down-regulating the expression of LPAR3.