miR-203a-3p靶向AKT2增强骨肉瘤细胞的放射敏感性

miR-203a-3p enhances radiosensitivity of osteosarcoma cells by targeting AKT2

目的:研究miR-203a-3p对骨肉瘤MG-63细胞凋亡和放射敏感性的影响及潜在作用机制。方法:qRT-PCR检测AKT2 mRNA的表达水平及不同放射剂量(0、2、4、6、8 Gy)照射后MG-63细胞中miR-203a-3p的表达水平,克隆形成实验检测不同放射剂量处理后细胞存活分数,流式细胞术测定MG-63细胞凋亡率,Western blot检测AKT2蛋白表达,双荧光素酶报告系统验证miR-203a-3p和AKT2的调控关系。结果:随着放射剂量的增加,骨肉瘤细胞MG-63中miR-203a-3p的表达量被显著抑制(P <0.05);过表达miR-203a-3p可增加MG-63细胞的放射敏感性,促进MG-63细胞凋亡;双荧光素酶报告系统结果显示,miR-203a-3p靶向负调控AKT2的表达;抑制AKT2表达可增强骨肉瘤MG-63细胞的放射敏感性;过表达AKT2逆转了上调miR-203a-3p对MG-63细胞的放射增敏作用,抑制MG-63细胞凋亡。结论:miR-203a-3p通过靶向抑制AKT2表达增强骨肉瘤MG-63细胞的放射敏感性,促进肿瘤细胞凋亡。miR-203a-3p有望成为骨肉瘤临床放射增敏靶点。

Objective:To investigate the effect of miR-203a-3p on apoptosis and radiosensitivity of osteosarcoma MG-63 cells and its potential mechanism.Methods:qRT-PCR was used to detect the expression level of AKT2 mRNA and the expression of miR-203a-3p in MG-63 cells treated with different doses of radiation(0,2,4,6,8 Gy).Colony formation assay was carried out to measure the survival fraction of MG-63 cells treated with different doses of radiation.Flow cytometry was used to detect the apoptotic rate of MG-63 cells.Western blot was applied to detect the expression level of AKT2 protein.And dual-luciferase reporter assay system was implemented to verify the relationship between miR-203a-3p and AKT2.Results:With the increase of radiation dose,the expression of miR-203a-3p in osteosarcoma MG-63 cells was significantly inhibited(P<0.05).Overexpression of miR-203a-3p enhanced the radiosensitivity and promoted the apoptosis of MG-63 cells.The results of dual-luciferase reporter assay system suggested that miR-203a-3p targeted and negatively regulated the expression of AKT2.Inhibition of AKT2 enhanced the radiosensitivity of osteosarcoma MG-63 cells.Overexpression of AKT2 reversed the radiosensitization of MG-63 cells when miR-203a-3p was up-regulated and inhibited the apoptosis of MG-63 cells.Conclusion:miR-203a-3p enhances radiosensitivity and promotes apoptosis of osteosarcoma MG-63 cells by targeting AKT2.miR-203a-3p is expected to be a target for clinical radiosensitization of osteosarcoma.