基础医学与临床 ›› 2021, Vol. 41 ›› Issue (7): 1013-1017.

• 研究论文 • 上一篇    下一篇

下调脂滴包被蛋白2抑制人胃癌细胞系NCI-N87增殖

刘炼玲1,李勇2   

  1. 1. 重庆市万州区人民医院
    2. 重庆大学附属三峡医院
  • 收稿日期:2020-09-04 修回日期:2021-01-24 出版日期:2021-07-05 发布日期:2021-06-17
  • 通讯作者: 李勇 E-mail:27057681@qq.com
  • 基金资助:
    重庆市自然科学基金面上项目

Down-regulation of perilipin 2 inhibits proliferation of human gastric cancer cell line NCI-N87

  • Received:2020-09-04 Revised:2021-01-24 Online:2021-07-05 Published:2021-06-17

摘要: 目的 研究脂滴包被蛋白2(perilipin2)在胃癌中的表达及对癌细胞增殖、侵袭及迁移的影响及其机制。方法 用人正常胃黏膜细胞系GES-1和人胃癌细胞系NCI-N87,将NCI-N87细胞分为对照组(不作任何处理)、NC-shRNA组(转染10 μg NC-shRNA重组慢病毒质粒)、perilipin2-shRNA组(转染10 μg perilipin2-shRNA重组慢病毒质粒)。用CCK8法检测细胞增殖率;用Transwell小室法检测细胞侵袭能力;用划痕实验检测细胞迁移能力;用Western blot法检测细胞中perilipin2、p62、Kelch样ECH相关蛋白1(keap1)、核转录因子E2相关因子2(Nrf2)通路相关蛋白表达。结果 NCI-N87细胞中的perilipin2蛋白表达量显著高于GES-1细胞(P<0.05);6、12、24、36和48 h时,perilipin2-shRNA组NCI-N87细胞增殖率较对照组、NC-shRNA组低(P<0.05);perilipin2-shRNA组穿过小室膜细胞数较对照组、NC-shRNA组低(P<0.05);perilipin2-shRNA组划痕间距较对照组、NC-shRNA组高(P<0.05);perilipin2-shRNA组NCI-N87细胞中的perilipin2、p62、keap1、Nrf2蛋白表达量较对照组、NC-shRNA组低(P<0.05)。结论 perilipin2在胃癌中呈高表达,下调perilipin2表达能够抑制NCI-N87细胞增殖、侵袭及迁移,其机制可能与p62-keap1-Nrf2通路受阻有关。

关键词: 脂滴包被蛋白2, 胃癌, 表达水平, 细胞增殖, 细胞侵袭, 细胞迁移, p62-keap1-Nrf2通路

Abstract: Objective To investigate the expression of perilipin 2 in gastric cancer and its effect on the proliferation, invasion and migration of cancer cells. Methods Human normal gastric mucosa cell line GES-1 and human gastric cancer cell line NCI-N87 were selected. NCI-n87 cells were divided into three groups. control group received no treatment, NC-shRNA group was transfected with 10μg NC-shRNA recombinant lentivirus plasmid, and perilipin2-shRNA group was transfected with 10μg perilipin2-shRNA recombinant lentivirus plasmid. The proliferation rate, invasion ability and migration ability of NCI-N87 cells in each group was detected by CCK8 method, Transwell chamber and scratch test, respectively. Western blot was used to detect the expression levels of perilipin2 and p62- kelch-like ECH-associated protein 1 (keap1) - nuclear factor E2-related factor-2 (Nrf2) pathway related proteins in NCI-N87 cells in each group. Results The expression of perilipn2 protein in NCI-N87 cells was significantly higher than that in GES-1 cells (P<0.05). After treatment for 6、12、24、36 and 48 h, proliferation rate of NCI-N87 cells in perilipin2-shRNA Group was lower than that in control and NC-shRNA group (P<0.05). The number of NCI-N87 cells crossing membrane in perilipin2-shRNA Group was lower than that in control and NC-shRNA group (P<0.05). The scratch space in perilipin2-shRNA Groupwas wider than that in control and NC-shRNA group (P<0.05). The protein expression of perilipn2, p62, keap1 and Nrf2 in NCI-N87 cells of perilipin2-shRNA Group was lower than that of control and NC-shRNA group (P<0.05). Conclusions Perilipin2 is highly expressed in gastric cancer cells, and knockdown of perilipin2 expression can inhibit the proliferation, invasion and migration of gastric cancer cells, and its mechanism may be related to the obstruction of the p62-keap1-Nrf2 pathway.

Key words: Perilipin2, Gastric cancer, Expression level, Cell proliferation, Cell invasion, Cell migration, p62-keap1-Nrf2 pathway