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中国应用生理学杂志 ›› 2021, Vol. 37 ›› Issue (5): 490-494.doi: 10.12047/j.cjap.6092.2021.042

• 研究论文 • 上一篇    下一篇

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*

张自新, 陆海洋, 杨治花, 刘长虎, 詹文华, 折虹, 赵仁   

  1. 宁夏医科大学总医院放疗科, 银川 宁夏 750004
  • 收稿日期:2020-05-14 修回日期:2021-01-24 出版日期:2021-09-28 发布日期:2021-11-24
  • 通讯作者: Tel: 13995218598, 13895617316; E-mail: zhaorendocter@126.com, ethan_004@163.com
  • 基金资助:
    *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)

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

ZHANG Zi-xin, LU Hai-yang, YANG Zhi-hua, LIU Chang-hu, ZHAN Wen-hua, ZHE Hong, ZHAO Ren   

  1. Department of Radiotherapy of General Hospital Affiliated the Ningxia Medical University, Yinchuan 750004, China
  • Received:2020-05-14 Revised:2021-01-24 Online:2021-09-28 Published:2021-11-24

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。

关键词: 复制蛋白A1, 鼻咽癌, 侵袭, 迁移, 细胞周期

Abstract: 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.

Key words: RPA1, nasopharyngeal carcinoma, invasion, migration, cell cycle

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