bax加强表达对Raji细胞程序性死亡效应

EFFECTS OF bax GENE OVEREXPRESSION ON PROGRAMMED CELL DEATH OF BURKITT LYMPHOMA RAJI CELL LINE

目的：研究ｂａｘ过量表达对Ｂｕｒｋｉｔ淋巴瘤细胞株细胞程序性死亡的效应及其机理。方法：亚克隆构建可过量表达ｂａｘα的真核表达载体，通过脂质体转染法导入Ｒａｊｉ细胞株中。ＰＣＲ及免疫荧光定量鉴定ｂａｘα在Ｒａｊｉ细胞株中的表达；流式细胞仪检测凋亡率。结果：成功构建ｐＳＦＦＶｂａｘαＳ和ｐＳＦＦＶｂａｘαＡＳ两种质粒，转染Ｒａｊｉ细胞后，测得转染ｐＳＦＦＶｂａｘαＳ的Ｒａｊｉ细胞的ｂａｘα表达量明显高于转染空载ｐＳＦＦＶｎｅｏ质粒的Ｒａｊｉ细胞株。φ（胎牛血清）＝０．０１诱导两种Ｒａｊｉ细胞４８ｈ，Ｒａｊｉｂａｘα的凋亡率为４８．６％，Ｒａｊｉ０的凋亡率为６．４％。结论：ｂａｘα在Ｒａｊｉ细胞中的加强表达可明显促进细胞的凋亡。

Objective:To investigate the effects and mechanism of bax overexpression on the programmed cell death of Burkitt lymphoma Raji cell line. Method:The mammalian expression vector pSFFVbax αneo, which overexpressed protein bax was constructed by subcloning. The plasmid was transfected into Raji cells by liposomemediated transfection. The expression of bax α was determined by PCR and immunoinfluoresence staining and flow cytometric analysis. Results:pSFFVbax αS and pSFFVbax αAS were constructed successfully. bax α expression in Rajibax α was apparently higher than that of Raji cells as control. The apoptosis rate of Rajibax α cells was 48.6% while Raji cells was 6.4% after being induced by 1% FCS+RPMI 1640. Conclusion: Overexpression of bax in Raji cells clearly enhance apoptosis of the cells.