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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。
To investigate whether alternative mRNA splicing of ERCC1 occurs after treatment of lung and cervical cancer cells with cisplatin, the total RNA of treated tumor cells and control cells was subjected to RT-PCR for the purpose of identifying the splicing isoforms TA bands were cloned and sequenced. As a result, two new ERCC1 mRNA splicing isoforms were found, in which the isoforms lacking exons 7-9 were ubiquitously found in many of the cancer cells tested, and their proportion in cisplatin treatment Will increase with the increase of cisplatin concentration. Changes in this isoform reveal that cells may regulate the function of ERCC1 by alternative pre-mRNA splicing.