长链非编码RNA-PVT1在顺铂诱导DNA损伤修复中的作用机制

Possible mechanism of lnc RNA-PVT1 on cisplatin-induced DNA damage and repair

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 and possible mechanism of lncRNA-PVT1 on cisplatin-induced DNA damage and repair. Methods The cisplatin-induced DNA damage model was established using A549 cells （non small cell lung cancer cell line）, and PVT1 expression was then detected by qRT-PCR. After silencing PVT1 with siPVT1, nucleotide excision repair （NER） related genes of A549 cells were determined by qRT-PCR and Western blot. The interaction between PVT1 and ERCC1 was detected by RNA immunoprecipitation （RIP） assay. Results PVT1 expression of A549 cells was dramatically decreased after treatment with cisplatin. Knockdown of PVT1 resulted in overexpression of γH2AX and downexpression of key NER-related genes including ERCC1 and DDB2 in A549 cells. RIP assay showed that PVT1 was interacted with ERCC1. Conclusion PVT can facilitate cisplatin-induced DNA damage and the repair via modulating NER pathway.