舒尼替尼调控miR-143对人肾癌细胞增殖、侵袭的影响

目的探讨miR-143在肾癌细胞中的作用、舒尼替尼对miR-143表达的影响及与细胞增殖、侵袭的关系。方法培养人肾癌786-O细胞株，分为对照组、miR-143组、舒尼替尼组、共同作用组。对照组细胞转染阴性对照序列，miR-143组细胞转染miR-143序列，舒尼替尼组细胞用舒尼替尼处理48 h，共同作用组细胞转染miR-143序列并用舒尼替尼处理48 h。MTT法检测细胞的增殖，Transwell检测其侵袭，qRT-PCR检测miR-143RNA表达水平，Western blotting检测DNA甲基转移酶3B（DNMT3B）、p-Akt-S473蛋白表达量。结果成功构建稳定、高表达miR-143的人肾癌786-O细胞株，miR-143转染组细胞miR-143表达量增高（P < 0.01）。舒尼替尼作用于786-O细胞48 h的IC₅₀值为6 μmol/L，选取该值作为后续实验用药浓度。786-O细胞经舒尼替尼作用48 h后miR-143表达量升高（P < 0.01）。miR-143组和舒尼替尼组细胞存活率、细胞侵袭数、DNMT3B和p-AktS473蛋白表达量均低于对照组，且高于共同作用组，差异均有统计学意义（P < 0.05~P < 0.01）。结论miR-143可以降低786-O细胞增殖、侵袭能力，这可能与miR-143降低DNMT3B、p-Akt的表达有关。舒尼替尼可能通过上调miR-143水平抑制786-O细胞的增殖与侵袭。

Effect of regulation of miR-143 by sunitinib on the proliferation and invasion of human renal carcinoma cells

ObjectiveTo investigate the role of miR-143 in renal cell carcinoma, the effect of sunitinib on the expression of miR-143 and its association with cell proliferation and invasion.MethodsThe human renal carcinoma 786-O cells were divided into control group, miR-143 group, sunitinib group and co-action group.The negative control sequence was transfected into the cells of control group, the miR-143 sequence was transfected into the cells of miR-143 group, sunitinib group cells were treated with sunitinib for 48h, the co-acting group was transfected miR-143 sequence and treated with sunitinib for 48h.Proliferation and invasion of 786-O cells were detected by MTT and transwell experiment, miR-143 RNA expression was detected by qRT-PCR, DNA methyltrans-ferase3B(DNMT3B).The expressions of p-Akt-S473 protein were detected by Western blotting.ResultsThe human renal cancer 786-O cells with stable and high expression of miR-143 were successfully constructed.The expression of miR-143 was increased in the miR-143 transfected group(P < 0.01).The IC₅₀ value in 786-O cells with the treatment of sunitinib for 48 h was 6 μmol/L, which was selected as the drug concentration in the subsequent experiment.The expression of miR-143 in 786-O cells was increased after the treatment of sunitinib for 48 h(P < 0.01).Cell survival rate, cell invasion number, expressions of DNMT3B and p-Akt-S473 protein in miR-143 and sunitinib group were all lower than control group and higher than co-action group(P < 0.05 to P < 0.01).ConclusionsmiR-143 reduced the proliferation and invasion capacity of 786-O cells, which may be related to the decreased expression of DNMT3B and p-Akt regulated by miR-143.Sunitinib may inhibit the proliferation and invasion of 786-O cells by upregulating miR-143.