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

Zhongguo Ying Yong Sheng Li Xue Za Zhi. 2021 Sep;37(5):490-494. doi: 10.12047/j.cjap.6092.2021.042.
[Article in Chinese]

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.

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。.

Keywords: RPA1; cell cycle; invasion; migration; nasopharyngeal carcinoma.

MeSH terms

  • Apoptosis
  • Cell Cycle
  • Cell Division
  • Cell Line, Tumor
  • Cell Proliferation
  • Down-Regulation
  • Gene Silencing
  • Humans
  • Nasopharyngeal Carcinoma* / genetics
  • Nasopharyngeal Neoplasms* / genetics
  • Replication Protein A / genetics*

Substances

  • RPA1 protein, human
  • Replication Protein A