下调脂滴包被蛋白2抑制人胃癌细胞系NCI-N87增殖

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

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.重庆市万州区人民医院消化内科,重庆 404000;
2.重庆大学附属三峡医院消化内科,重庆 404000

收稿日期:2020-09-04 修回日期:2021-01-26 出版日期:2021-07-05 发布日期:2021-06-17
通讯作者: *46960864@qq.com
基金资助:
重庆市自然科学基金(cstc2017jcyj-mxmX0868)

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

LIU Lian-ling¹, LI Yong^2*

1. Department of Gastroenterology, Wanzhou District People's Hospital, Chongqing 404000;
2. Department of Gastroenterology, Chongqing University Three Gorges Hospital,Chongqing 404000, China

Received:2020-09-04 Revised:2021-01-26 Online:2021-07-05 Published:2021-06-17
Contact: *46960864@qq.com

摘要/Abstract

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。方法采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组宽(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, 肿瘤生物学行为, 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 10 μg NC-shRNA recombinant lentivirus plasmid, and perilipin2-shRNA group was transfected with 10 μg perilipin2-shRNA recombinant lentivirus plasmid. The proliferation rate, invasion ability and migration ability of NCI-N87 cells in each group were detected by CCK8 method, Transwell chamber and scratch test, respectively. Western blot was used to detect the expression levels of perilipin2, 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 level 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 less than that in control and NC-shRNA group(P＜0.05). The scratch space in perilipin2-shRNA group was 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, tumor biological behavior, p62-keap1-Nrf2 pathway

中图分类号:

刘炼玲, 李勇. 下调脂滴包被蛋白2抑制人胃癌细胞系NCI-N87增殖[J]. 基础医学与临床, 2021, 41(7): 1013-1017.

LIU Lian-ling, LI Yong. Down-regulation of perilipin 2 inhibits proliferation of human gastric cancer cell line NCI-N87[J]. Basic & Clinical Medicine, 2021, 41(7): 1013-1017.

参考文献

[1]章盈婧, 于健春. 肿瘤相关巨噬细胞在胃癌发展与治疗中的研究进展[J]. 基础医学与临床, 2019,12:885-886.
[2]徐惠绵, 王鑫. 我国胃癌诊治临床研究现状与展望[J]. 中华胃肠外科杂志, 2020, 23:109-114.
[3]Libby AE, Bales ES, Jenifer M, et al. Perilipin-2 deletion promotes carbohydrate-mediated browning of white adipose tissue at ambient temperature[J]. J Lipid Res, 2018, 59:2563-2564.
[4]Jin, Yanjie, Tan, et al. Reactive oxygen species induces lipid droplet accumulation in hepG2 cells by increasing perilipin 2 expression.[J]. Int J Mol Sci, 2018,4:662-664.
[5]Cao Q, Ruan H, Wang K, et al. Overexpression of PLIN2 is a prognostic marker and attenuates tumor progression in clear cell renal cell carcinoma[J]. Int J Oncol, 2018,5:56-60.
[6]Conte M, Armani A, Conte G, et al. Muscle-specific perilipin2 down-regulation affects lipid metabolism and induces myofiber hypertrophy[J]. J Cachexia Sarcopenia Muscle, 2019, 10:789-790.
[7]Saliba-Gustafsson P, Pedrelli M, Werngren O, et al. The lipid-droplet associated protein perilipin 2 (PLIN2) plays a central role in lipid accumulation and cholesterol efflux via effects on LXR signaling in human macrophages[J]. Atherosclerosis, 2018, 275:2563-2567.
[8]Shen Z, Wang Y, Su Z, et al. Activation of p62-keap1-Nrf2 antioxidant pathway in the early stage of acetaminophen-induced acute liver injury in mice[J]. Chem Biol Interact, 2018, 282:1235-1237.
[9]Choi Y, Jiang Z, Shin WJ, et al. SFTS virus NSs interacts with TRIM21 to activate the p62-Keap1-Nrf2 pathway[J]. J Virol, 2020, 94:1684-1688.
[10]Yasuji K, Kazuhisa N, Toshihiko S, et al. Tu1479-Activation of the P62-KEAP1-NRF2 pathway in patients with non-B non-C hepatocellular carcinoma and type 2 diabetes[J]. Gastroenterology, 2018, 154:1232-1234.

[1]	刘磊, 李世宝, 张好良. 沉默lnc-SLC2A12-10:1抑制人胃癌细胞系AGS增殖、迁移和侵袭[J]. 基础医学与临床, 2022, 42(7): 1071-1076.
[2]	陈希, 庞海玉, 秦春迪, 郭元红, 杨爱明, 赖雅敏. 学习金字塔模型理论的短期半浸入式培训提高识别早期胃癌能力[J]. 基础医学与临床, 2022, 42(4): 696-700.
[3]	牛世伟, 苏韫, 龚红霞, 李婷婷, 曾元丁, 张晗, 聂蓬. 胃癌患者血清IL-1的升高和癌组织JAK2/STAT3信号通路的激活[J]. 基础医学与临床, 2022, 42(3): 401-405.
[4]	侯楠, 刘源, 高俊, 王晶. miR-141-3p通过靶向PDCD1抑制人胃癌细胞系AGS的迁移与侵袭[J]. 基础医学与临床, 2021, 41(9): 1297-1302.
[5]	苏鹏飞, 蒋林, 章盈婧, 郭博, 马志强, 周炜洵, 于健春. 综合治疗79岁进展期胃癌1例[J]. 基础医学与临床, 2021, 41(7): 1043-1046.
[6]	鲍美美徐雯王超王佳苗小芬. 石蒜碱抑制胃癌细胞系MGC-803增殖、迁移和侵袭[J]. 基础医学与临床, 2021, 41(12): 1767-1773.
[7]	朱萌张宁陶伟. 人胃癌细胞系AGS外泌体miR-106a对腹膜间皮细胞系HMrSV5表型的影响[J]. 基础医学与临床, 2021, 41(11): 1564-1569.
[8]	杨屹立, 何学元, 潘新民, 马建勋. miR-205-5p抑制体外胃癌细胞系增殖、迁移和侵袭[J]. 基础医学与临床, 2020, 40(8): 1068-1075.
[9]	李春波, 苏韫, 龚红霞, 薛轩, 李婷婷, 牛世伟, 聂蓬. IL-17通过NF-κB信号通路参与胃癌发生发展的免疫调控[J]. 基础医学与临床, 2020, 40(7): 923-928.
[10]	王小娟, 李小丽, 金正旭, 张德奎. GDNF与消化系统肿瘤关系研究进展[J]. 基础医学与临床, 2020, 40(7): 986-989.
[11]	尹超云, 张垚, 陈振威, 张安伟, 张旋风, 张日亭, 步雪峰. 重组新城疫病毒 IL29 抑制人胃癌细胞系BGC的增殖与迁移[J]. 基础医学与临床, 2020, 40(3): 368-373.
[12]	陈国雁, 冯晓波. 抑制miR-621表达促进人胃癌细胞系增殖、迁移和侵袭[J]. 基础医学与临床, 2020, 40(11): 1500-1505.
[13]	史济华徐雪崔海梦张攀罗庆锋许乐. 血清菱形结构域蛋白、胃蛋白酶原和癌胚抗原对胃癌的早期诊断价值[J]. 基础医学与临床, 2019, 39(9): 1316-1319.
[14]	仇建玲刘军. 替吉奥分别联合紫杉醇或奥沙利铂一线治疗晚期胃癌的疗效及安全评价[J]. , 2019, 39(12): 1752-1755.
[15]	王欢何丛吕农华. 胃肠道微生态与幽门螺杆菌相关疾病的研究进展[J]. 基础医学与临床, 2018, 38(11): 1611-1614.