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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 alteration of Bim mRNA alternative splicing isoforms in six different tumor cell lines treated with cisplatin. METHODS: Lung cancer cells (A549, H1299), human embryonic kidney cells (293FT) and cervical cancer cells (SiHa, CaSki, C33A) were treated with a gradient of cisplatin. Total RNA was extracted from each cell. Agarose gel electrophoresis and gel image analysis software detection and analysis of Bim mRNA isoforms composition ratio and the relative level of change. RESULTS: Six isoforms of BimEL, BimL, BimS, Bimα2, Bimα4 and Bimβ7 were detected in all six cell lines after cisplatin treatment. With the increase of cisplatin concentration, BimL isoforms Body proportion and expression levels have increased significantly. CONCLUSIONS: Cisplatin may up-regulate the expression of Bim gene through transcription and alternative splicing of mRNA to promote cell apoptosis. BimL is mainly up-regulated isoform.