摘要
[目的]探究miR-494对骨肉瘤细胞增殖和侵袭的调控作用及机制。[方法]对骨肉瘤MG-63细胞采用瞬时转染法上调/下调miR-494水平,采用RT-PCR法检测mRNA表达水平,CCK-8法检测细胞的增殖能力,Transwell法检测细胞的侵袭能力,划痕实验检测细胞迁移能力,Western blot检测目标蛋白表达水平。[结果]瞬时转染后随时间推移,两组miR-494的表达均显著增加(P<0.05),转染后12、18以及24 h时,转染组的miR-494表达情况均高于对照组(P<0.05)。随时间推移,两组细胞增殖均显著增加(P<0.05),24、48以及72 h时,转染组细胞增殖水平均显著低于对照组(P<0.05)。转染组在视野下的平均细胞数、细胞跨膜数量以及细胞迁移率均显著低于对照组(P<0.05)。转染组CDK6的mRNA以及蛋白表达均显著低于对照组(P<0.05)。[结论]miR-494的高表达能够抑制骨肉瘤细胞的增殖,并且能够导致CDK6的mRNA以及蛋白质表达水平降低。
[Objective]To explore the regulation and mechanism of miR-494 on proliferation and invasion of osteosarcoma cells.[Methods]By using transient transfection method,the level of miR-494 in osteosarcoma MG-63 cells were up-regulated,or down-regulat⁃ed.The mRNA expression level was assessed by RT-PCR,cell proliferation capacity by CCK-8 assay,cell invasion ability by Transwell test,cell migration ability by scratch test and target protein expression level was detected by western blot.[Results]With the passage of time after transient transfection,the expression of miR-494 in the two groups increased significantly(P<0.05).At 12h,18h and 24h after transfection,the expression of miR-494 in the transfection group was higher than that of the control group(P<0.05).In addition,the cell proliferation of the two groups increased significantly over time(P<0.05).At 24 h,48 h and 72 h,the cell proliferation level of the transfec⁃tion group was significantly lower than that of the control group(P<0.05).The average cell number,cell transmembrane number and cell migration rate of the transfection group under the field of view were significantly lower than those of the control group(P<0.05).The mRNA and protein expression of CDK6 in the transfection group were significantly lower than those in the control group(P<0.05).[Conclusion]The high expression of miR-494 can inhibit the proliferation of osteosarcoma cells,and lead to the reduction of CDK6 mRNA and protein expression levels.
作者
张雷
段广超
吴智辉
ZHANG Lei;DUAN Guang-chao;WU Zhi-hui(Department of Spine Surgery,The First Peoples Hospital of Shangqiu City,Shangqiu 476100,China)
出处
《中国矫形外科杂志》
CAS
CSCD
北大核心
2021年第21期1974-1977,共4页
Orthopedic Journal of China
