miR-107抑制胶质瘤细胞的体外增殖、迁移和侵袭

目的探讨miR-107对胶质瘤细胞增殖、迁移和侵袭的调控机制。方法运用RT-qPCR检测人正常的星形胶质细胞系NHA、神经胶质瘤细胞系U87、A172、U251中miR-107和FOXK1的表达;将细胞分为miR-NC组(转染miR-NC)、miR-107组(转染miR-107 mimics)、si-NC组(转染si-NC)、si-FOXK1组(转染si-FOXK1)、miR-107+pcDNA3.1组(共转染miR-107 mimics和pcDNA3.1)和miR-107+pcDNA3.1-FOXK1组(共转染miR-107 mimics和pcDNA3.1-FOXK1);用脂质体法分别转染至U87细胞;CCK-8法检测细胞的增殖;Transwell小室实验检测细胞的迁移和侵袭;Western blot检测细胞中FOXK1的蛋白表达;双荧光素酶报告基因检测实验检测细胞的荧光活性。结果与正常的星形胶质细胞NHA相比,神经胶质瘤细胞U87、A172、U251中miR-107表达明显下调,FOXK1表达明显上调(P<0.05);过表达miR-107、敲减FOXK1均可抑制U87细胞的增殖、迁移和侵袭;miR-107可抑制野生型FOXK1的细胞荧光活性,并负向调控FOXK1的表达;过表达FOXK1可逆转miR-107对U87细胞增殖迁移侵袭的抑制作用。结论 miR-107抑制胶质瘤细胞增殖、迁移和侵袭的作用机制可能与靶向负调控FOXK1有关,将可为胶质瘤的诊断和治疗提供靶向治疗的依据。

miR-107 inhibits the proliferation,migration and invasion of glioma cells in vitro

Objective To investigate the regulation mechanism of miR-107 on proliferation, migration and invasion of glioma cells. Methods RT-qPCR was used to detect the expression of miR-107 and FOXK1 in human normal astrocytes NHA, glioma cell lines U87, A172 and U251. miR-NC(miR-NC group), miR-107 mimics(miR-107 group), si-NC(si-NC group), si-FOXK1(si-FOXK1 group), miR-107+pcDNA3.1(miR-107 mimics and pcDNA3.1 group), miR-107 +pcDNA3.1-FOXK1(miR-107 mimics and pcDNA3.1-FOXK1 group), were transfected respectively into U87 cells by liposome method. The proliferation of the cells was detected by CCK-8 method.The migration and invasion of the cells were detected by Transwell chamber assay. The protein expression of FOXK1 in the cells was detected by Western blot.The fluorescence activity of the cells was detected by the dual luciferase reporter assay. Results Compared with normal astrocyte NHA, the expression of miR-107 was down-regulated and the expression of FOXK1 was up-regulated in glioma cell lines U87, A172 and U251(P<0.05). Over-expression ofmiR-107 and knockdown of FOXK1 inhibited U87 cell proliferation, migration and invasion. MiR-107 can inhibit the cytosolic activity of wild-type FOXK1 and negatively regulate the expression of FOXK1;over-expression of FOXK1 can reverse the inhibitory effect of miR-107 on proliferation and migration of U87 cells. Conclusions The mechanism of miR-107 inhibiting proliferation, migration and invasion of glioma cells may be related to the targeted negative regulation of FOXK1, which will provide a basis for the diagnosis and targeted therapy of glioma.