ERCC1新的mRNA剪接异构体及其在顺铂处理细胞中的变化

New mRNA splicling isoforms of ERCC1 and their change in cells after treatment with cisplatin

为了考察用顺铂处理肺癌和宫颈癌细胞后,ERCC1的mRNA选择性剪接是否发生变化,通过提取处理的肿瘤细胞和对照细胞的总RNA,进行RT-PCR检测,对出现的剪接异构体目的条带进行TA克隆、测序鉴定。结果发现了2个新的ERCC1的mRNA剪接异构体,其中缺失外显子7-9的异构体在所检测的多种癌细胞中普遍存在,并且在顺铂处理下,其所占比例会随顺铂浓度的升高而增加。该异构体的变化揭示细胞可能会通过pre-mRNA选择性剪接调控ERCC1的功能。

In order to find out whether there is any change of mRNA alternative splicing isoforms of ERCC1 in lung cancer and cervical cancer cells after treatment with different dose of cisplatin,total RNA of treated cells and control cells were abstracted and subjected to RT-PCR to test the spliced transcripts and their ratios.The target bands were cloned into TA vector to sequence.Two new mRNA splicing isoforms of ERCC1 were found.The shortest isoform which lost exons 7-9was found in almost checked cell lines.Under the treatment of cisplatin,the ratio of the shortest isoform increased when the dose of cisplatin went up.Based on the reported data,it is suggested that the proteins coded by these two new mRNA isoforms will not combine with XPF to function NER（nucleotide excision repair）.These results showed that cells can utilize the alternative splicing to control the level and ratio of the different mRNA isoforms to handle the function of ERCC1.