顺铂处理后几种癌细胞中Bim基因mRNA异构体的变化

The Change of mRNA Isomer of Bim Gene in Several Cancer Cells under the Cisplatin Treatment

目的:探讨不同浓度顺铂处理6种肿瘤细胞后,细胞中Bim基因mRNA选择性剪接异构体的表达及变化规律。方法:用梯度浓度的顺铂处理肺癌细胞(A549、H1299)、人胚肾细胞(293FT)、宫颈癌细胞(SiHa、CaSki、C33A),提取各细胞的总RNA,用半定量RT-PCR、琼脂糖凝胶电泳及凝胶图像分析软件检测分析Bim基因mRNA异构体的组成比例和相对水平的变化。结果:用顺铂处理后,在6种细胞中都检测到了BimEL、BimL、BimS、Bimα2、Bimα4和Bimβ7共6种mRNA异构体;随着顺铂处理浓度的增高,各细胞中BimL异构体所占比例和表达水平都有明显的增加。结论:顺铂可能通过转录和mRNA选择性剪接两种机制上调Bim基因的表达,从而促进细胞凋亡;且BimL是主要被上调的异构体。

Objective： To investigate the expression and variation of mRNA isoforms of Bim gene in 6 kinds of tumor cells after treaed with different concentration of cisplatin. Method： The cells of lung cancer（ A549 and H1299）,cervical cancer（ SiHa,CaSki and C33A） and human embryonic kidney cells 293 FT were treated by gradient dose of cisplatin. Total RNAs of the control cells and the treated cells were abstracted. The mRNA isoforms were detected by semiquantitative RT-PCR,agarose gel electrophoresis and Gel image analysis software to detect and analyze the change of composing proportion and relative level of mRNA isomer of Bim gene. Result： 6 mRNA isoforms of BimEL,BimL,BimS,Bimα2,Bimα4 and Bimβ7 were detected. With the increase of the concentration of cisplatin,the proportion and expression level of BimL isomer increased significantly. Conclusion： After cisplatin treatment,mRNA level of Bim gene is up-regulated by transcription and pre-mRNA alternative splicing to promote apoptosis in all detected cells,BimL is the dominant mRNA isoform.