论文部分内容阅读
目的研究长链非编码RNA-PVT1对顺铂诱导DNA损伤修复的影响及其作用机制。方法建立顺铂诱导非小细胞肺癌细胞株A549细胞DNA损伤模型,用荧光定量PCR(qRT-PCR)检测PVT1表达。通过siPVT1沉默A549细胞中PVT1表达,用qRT-PCR和Western blot检测核苷酸切除修复(NER)相关基因表达。用RNA免疫共沉淀(RIP)检测PVT1与ERCC1相互作用。结果顺铂处理A549细胞明显下调PVT1表达。沉默PVT1后A549细胞中γH2AX表达显著增高,而NER途径中关键基因如ERCC1、DDB2表达明显降低。RIP实验表明PVT1可与ERCC1蛋白直接相互作用。结论 PVT1可能通过调节NER途径促进顺铂诱导DNA损伤修复。
Objective To investigate the effect of long-chain non-coding RNA-PVT1 on cisplatin-induced DNA damage repair and its mechanism. Methods The DNA damage model of non-small cell lung cancer cell line A549 was established by cisplatin. The expression of PVT1 was detected by qRT-PCR. The expression of PVT1 in A549 cells was silenced by siPVT1, and the expression of NER related genes was detected by qRT-PCR and Western blot. RNA immunoprecipitation (RIP) was used to detect the interaction of PVT1 with ERCC1. Results Cisplatin treatment of A549 cells significantly down-regulated PVT1 expression. The expression of γH2AX in A549 cells was significantly increased after silencing PVT1, whereas the expression of key genes such as ERCC1 and DDB2 in NER pathway was significantly decreased. RIP experiments show that PVT1 can directly interact with ERCC1 protein. Conclusion PVT1 may promote cisplatin-induced DNA damage repair through regulating NER pathway.