miR-19a过表达对口腔鳞癌细胞CAL27增殖和迁移的影响及其机制研究

Effect and mechanism of overexpression of miR-19a on the proliferation and migration of oral squamous cell carcinoma cell line CAL27

目的探究miR-19a对口腔鳞癌细胞增殖和迁移的影响,进一步探究其机制与GRK6介导的PKC表达变化是否有关。方法脂质体法向CAL27细胞分别转染miR-19a模拟物(mimic组)和对照随机序列(对照组),RT-PCR检测miR-19a水平,MTT法检测增殖能力,划痕实验检验CAL27各组细胞迁移能力。通过蛋白免疫印迹法检测GRK6和PKC的表达。结果转染后的CAL27细胞株miR-19a表达明显增加(P<0.05);与阴性对照组相比,miR-19a过表达的CAL27细胞增殖能力明显增强(P<0.05);划痕实验结果显示miR-19a过表达增强了CAL27细胞的迁移能力;与阴性对照组相比,miR-19a过表达组的GRK6蛋白水平下降,而PKC蛋白含量升高(均P<0.05)。结论miR-19a具有增强CAL27细胞增殖和迁移能力的作用,其机制可能与GRK6介导的PKC表达变化有关。

Objective To explore the effect of miR-19a on the proliferation and migration of oral squamous cell carcinoma cells,and further investigate whether its mechanism is related to the changes in PKC expression mediated by GRK6.Methods The liposome method was used to transfect miR-19a mimic(mimic group)and control random sequence(control group)into CAL27 cells,respectively.The levels of miR-19a were detected by RT-PCR.The proliferation ability was detected by MTT method,and the migration ability ofCAL27 cells in each group was tested by scratch test.The expression of GRK6 and PKC was detected by protein immunoblotting.Results The expression of miR-19a in the transfected CAL27 cell line significantly increased(P<0.05).Compared with the negative control group,the proliferation ability of CAL27 cells overexpressed with miR-19a was significantly enhanced(P<0.05).The scratch test results showed that overexpression of miR-19a enhanced the migration ability ofCAL27 cells.Compared with the negative control group,the GRK6 protein level in the miR-19a overexpression group decreased,while the PKC protein content increased(all P<0.05).Conclusion MiR-19a has the effect of enhancing the proliferation and migration ability of CAL27 cells,and its mechanism may be related to the changes in PKC expressionmediated by GRK6.