hsa-miR-30a在人骨肉瘤细胞U2-OS中的表达及其对细胞凋亡的影响

Expression of hsa-miR-30a in human osteosarcoma U2-OS cells and the effect on the apoptosis of U2-OS

目的:研究miR-30a在人骨肉瘤细胞U2-OS中的过表达对其凋亡的影响。方法:构建pEZX-MR04-pre-miR-30a真核表达载体,瞬时转染人骨肉瘤细胞株U2-OS,荧光显微镜及荧光定量PCR法(qRT-PCR)检测miRNA-30a在U2-OS中的表达,并采用流式细胞仪检测稳转株细胞的凋亡情况。结果:荧光显微镜下观察真核表达载体pEZX-MR04-pre-miR-30a转染后的U2-OS细胞,可见大量绿色荧光;qRT-PCR检测U2-OS中miR-30a的表达明显提高;流式细胞检测转染后的U2-OS细胞的凋亡明显增加。结论:miR-30a在人骨肉瘤细胞U2-OS过表达能促进其凋亡,miR-30a可能是U2-OS细胞潜在的抑癌基因。

Objective：To determine the expression of miR-30a in human osterosarcoma cell line U2-OS and the effects on the apoptosis of human U2-OS cells.Methods：pEZX-MR04-pre-miR-30a vector was constructed and transiently transfected into U2-OS cells.The expression of miRNA-30a in U2-OS cells was verified by fluorescence microscopy and quantitative real time PCR （qRT-PCR）.Flow cytometry was used to detect the effects of miR-30a on the apoptosis of U2-OS cells.Results：pEZX-MR04-pre-miR-30a vector was successfully transfected into U2-OS cells.Quantitative green positive cells were seen under fluorescence microscope,and qRT-PCR results indicated significant increase of miR-30a expression in U2-OS cell lines.Flow cytometry findings showed that miR-30a significantly promoted U2-OS cells apoptosis.Conclusion：Over-expression of miR-30a has significantly facilitated the apoptosis cells U2-OS,suggesting that miR-30a may be a potential tumor suppressor gene for human osteosarcoma.