Wrap53调控p53基因表达对人非小细胞肺癌H322细胞放射敏感性的研究  

Effect of expression of p53 gene regulated by Wrap53 on radiosensitivity of H322 cell in non-small cell lung cancer

作　　者：吴传高 叶兴发 刘晓青 魏婷芳 彭强 袁岩成 董柱清 wU Chuan-gao;YE Xing-fa;LU Xiao-qing;WEI Ting-fang;PENG Qiang;YUAN Yan-cheng;DONG Zhu-qing(Department of Radiotherapy,Mindong Hospital,Ningde 355000,China)

机构地区：[1]福建省宁德市闽东医院放疗科,宁德355000

出　　处：《中国肿瘤临床与康复》2020年第6期687-690,共4页Chinese Journal of Clinical Oncology and Rehabilitation

基　　金：福建省宁德市科学技术计划项目(20130124)。

摘　　要：目的探讨Wrap53调控p53基因表达对人非小细胞肺癌H322细胞放射敏感性的影响及分子机制。方法选取中国科学院上海细胞生物研究所细胞库的人非小细胞肺癌H322细胞(p53突变型),构建Wrap53-shRNA干扰质粒,将ph Wrap53-shRNA转染至H322细胞,RT-PCR、Western Blotting H322检测Wrap53、Mtp53的mRNA及蛋白的表达;克隆形成实验计算准阈剂量(Dq),平均致死剂量(D0)和照射2Gy时细胞的存活率(SF2)等放射敏感性参数值。采用流式细胞仪检测细胞周期变化。结果与未转染组和sh-NC组比较,ph Wrap53-shRNA组细胞Wrap53和Mtp53的mRNA和蛋白表达水平均下降;ph Wrap53-shRNA组D0、Dq值和SF2值均低于sh-NC组,差异均有统计学意义(均P<0.05),放射增敏比(SER)为(1.468±0.023)。与sh-NC组相比,ph Wrap53-shRNA组在G1期阻滞增加,且以24 h时最显著;Wrap53干扰后在放射16 h和24 h后的G2/M期阻滞明显减少,差异均有统计学意义(均P<0.05)。结论干扰Wrap53的表达增加了人非小细胞肺癌H322细胞的放射敏感性,可能与抑制Mtp53基因的表达和改变放射后的细胞周期时相有关,有望成为增加肿瘤细胞放射敏感性的一个潜在靶点。Objective To investigate the effect of expression of p53 gene regulated by Wrap53 on radiosensitivity of H322 cell in non-small cell lung cancer.Methods H322 cell(mutant p53)in nonsmall cell lung cancer was selected from Shanghai Institute of Biochemistry and Cell Biology Cell Band,Chinese Academy of Sciences.Wrap53 RNA interference was constructed.H322 cell was transfected by ph Wrap53-shRNA.Expression of Wrap53 and Mtp53 mRNA and protein was tested by RT-PCR and Western Blot Assay H322.Radiosensitivity parameters including Threshold dose(Dq),mean lethal dose(D0),and cell survival(SF2)when exposed to 2 Gy for statistical calculation was formed after the clones.Changes in cell cycle were detected by flow cytometry.Results Compared with untransfected group and sh-NC group,the expression of Wrap53 and Mtp53 mRNA and protein decreased in ph Wrap53-shRNA group(P<0.05).D0,Dq and SF2 were lower in ph Wrap53-shRNA group than in sh-NC group(P<0.05).Sensitizing enhancement ratio(SER)was 1.468±0.023.Compared to the sh-NC group,the increase of G1-arrested cells was more prominent in ph Wrap53-shRNA group especially at 24 h after radiation(P<0.05).In contrast,the G2/M-arrested cells obviously decreased in ph Wrap53-shRNA group at 16 h and 24 h,respectively after radiation(all P<0.05).Conclusion Silencing of Wrap53 expression can enhance the radiosensitivity of H322 cells effectively,which may be related to suppression of the expression of Mtp53 gene and altered cell cycle phases after radiation.It is expected to be a potential target for increasing the radiosensitivity of tumor cells.

关 键 词：Wrap53 MTP53 癌 非小细胞肺 RNA干扰 放射敏感性 细胞周期 

分 类 号：R734.2[医药卫生—肿瘤]