过表达PCL2基因促进人胶质瘤U87细胞增殖的作用研究

Over-expression of PCL2 gene promotes proliferation of human glioma U87 cells

目的:构建过表达多梳状蛋白2(PCL2)基因的重组腺病毒,观察其促进人胶质瘤细胞系U87增殖的作用。方法:构建PCL2重组腺病毒载体Ad5-E1-CMV-hPCL2-E3-CMV-eGFP,利用HEK293细胞包装制备腺病毒,感染U87细胞,应用倒置荧光显微镜观察感染效率;用Western Blot检测U87细胞内PCL2基因的蛋白表达水平; CCK8和集落形成实验检测细胞的增殖能力。结果:成功构建过表达PCL2基因的重组腺病毒,当MOI50∶1时,感染效率最高。Western Blot结果显示过表达PCL2基因后,PCL2蛋白表达水平较空白对照组和空载体组明显升高。CCK8和集落形成实验提示PCL2基因过表达促进U87细胞增殖。结论:PCL2基因过表达能够促进人胶质瘤U87细胞增殖。

Objective: Over-expression of Polycomb like 2( PCL2) gene was constructed to observe its effect on the proliferation of human glioma U87 cells. Methods: The recombinant adenovirus Ad5-E1-CMV-hPCL2-E3-CMV-eGFP,packed by HEK293 cell,was constructed and infected with human glioma U87 cells. The infection efficiency was observed by inverted fluorescence microscope. The protein expression of PCL2 gene in U87 cells was detected by Western Blot;CCK8 and colony formation experiment were used to detect cell proliferation. Results: When the over-expressed PCL2 gene MOI 50∶1 was successfully constructed,the infection efficiency was the highest. Western Blot results showed that the expression level of PCL2 protein was significantly higher than that of blank control group and empty vector group after over-expression of PCL2 gene. CCK8 and colony formation experiment suggested that over-expression of PCL2 gene promoted the proliferation of U87 cells. Conclusion: Over-expression of PCL2 gene could promote the proliferation of human glioma U87 cells.