RPA1低表达对辐射抵抗人鼻咽癌CNE-2R细胞侵袭迁移及细胞周期的影响  

Effects of RPA1 down-regulation on the invasion and migration and cell cycle of human nasopharyngeal carcinoma CNE-2R cells with radio resistance

作　　者：张自新[1] 陆海洋 杨治花[1] 刘长虎[1] 詹文华[1] 折虹[1] 赵仁[1] ZHANG Zi-xin;LU Hai-yang;YANG Zhi-hua;LIU Chang-hu;ZHAN Wen-hua;ZHE Hong;ZHAO Ren(Department of Radiotherapy of General Hospital Affiliated the Ningxia Medical University,Yinchuan 750004,China)

机构地区：[1]宁夏医科大学总医院放疗科,银川宁夏750004

出　　处：《中国应用生理学杂志》2021年第5期490-494,共5页Chinese Journal of Applied Physiology

基　　金：宁夏医科大学临床学院导师孵育项目(CXJH04);宁夏自然科学基金项目(2018AAC03159)。

摘　　要：目的:探讨复制蛋白A1(RPA1)沉默对人鼻咽癌CNE-2R细胞侵袭、迁移及细胞周期的影响。方法:采用shRNA技术构建RPA1低表达的CNE-2R细胞模型并通过RT-PCR和Western blot实验验证。选用空白对照组(CNE-2R)、阴性对照组(NC-shRNA)、RPA1低表达组(RPA1-shRNA)3组细胞完成后续实验,通过CCK8和克隆形成实验检测细胞增殖能力、Transwell实验检测侵袭能力、划痕实验检测迁移能力,流式细胞术检测细胞周期;Western blot实验检测Chk2、p-Chk2、Cdc25c和p-cdc25c蛋白的表达。结果:与CNE-2R和NC-shRNA组比较,RPA1-shRNA组细胞的RPA1mRNA和蛋白质均显著降低(P<0.01和<0.05);RPA1-shRNA组组细胞的增殖、侵袭、迁移能力显著下降(P均<0.05),细胞周期被阻滞在G2/M期(P<0.01);RPA1-shRNA组细胞Chk2、Cdc25c的表达低于CNE-2R和NC-shRNA组细胞(P<0.05),而p-Chk2、p-cdc25c的表达高于其它两组(P<0.05)。结论:RPA1低表达抑制辐射抵抗人鼻咽癌CNE-2R细胞的增殖、迁移以及使细胞周期阻滞于G2/M期。Objective:To investigate the effects of RPA1 silencing on the invasion,migration and cell cycle of human nasopharyngeal carcinoma CNE-2R cells.Methods:shRNA technology was used to construct CNE-2R cell lines with RPA1 low-expression,which were verified by RT-PCR and Western blotting.The following assays were performed using the three 3 groups:control group(CNE-2),negative control group(NC-shRNA)and RPA1 down-regulation group(RPA1-shRNA).The effects of RPA silence on the proliferation,invasion,migration,and cell cycle of CNE-2R cells were detected using Cell Counting Kit-8,clone formation experiment,Transwell,scratch test and flow cytometry,respectively.The expressions of Chk2,p-Chk2,Cdc 25c and p-cdc25c were tested by Western blot assay.Results:The expressions of RPA1 mRNA and protein in the RPA1-shRNA group were lower than those in the CNE-2 and NC-shRNA groups significantly(P<0.01 and 0.05).Compared with CNE-2 and NC-shRNA groups,the abilities of proliferation,invasion and migration of RPA1-shRNA group were decreased and the cell cycle in the RPA1-shRNA group was blocked in the G2/M phase(P<0.01).The expressions of Chk2 and Cdc25c in RPA1-shRNA group cells were lower than those in CNE-2R and NC-shRNA group cells(P<0.05),while the expressions of p-Chk2 and p-cdc25c were higher than those in the other groups(P<0.05).Conclusion:After RPA1 silenced,the proliferation and migration of radio resistant human nasopharyngeal carcinoma CNE-2R cells was inhibited,resulting in cell cycle arrested in the G2/M phase.

关 键 词：复制蛋白A1 鼻咽癌 侵袭 迁移 细胞周期 

分 类 号：R739.63[医药卫生—肿瘤]