吉林大学学报(医学版) ›› 2020, Vol. 46 ›› Issue (02): 266-273.doi: 10.13481/j.1671-587x.20200210

• 基础研究 • 上一篇    下一篇

LincRNA-p21敲减对胃癌细胞生长和转移的影响及其作用机制

徐建国1, 曹洪涛1, 张子龙1, 冀保妍2, 王春秋3, 李生栋1, 刘国庆1   

  1. 1. 青海省人民医院肿瘤外科, 青海 西宁 810001;
    2. 青海省人民医院肿瘤内科, 青海 西宁 810001;
    3. 青海省人民医院消化内科, 青海 西宁 810001
  • 收稿日期:2019-04-08 发布日期:2020-04-07
  • 通讯作者: 刘国庆,主任医师,硕士研究生导师(Tel:0971-8177911,E-mail:liuguoqing8853@163.com) E-mail:liuguoqing8853@163.com
  • 作者简介:徐建国(1982-),男,青海省乐都县人,副主任医师,医学硕士,主要从事胃肠道肿瘤基础与临床方面的研究。
  • 基金资助:
    青海省卫健委指导性科研项目资助课题(2018-wjzdx-12)

Effects of LincRNA-p21 knockdown on growth and metastasis of gastric cancer cells and their mechanisms

XU Jianguo1, CAO Hongtao1, ZHANG Zilong1, JI Baoyan2, WANG Chunqiu3, LI Shengdong1, LIU Guoqing1   

  1. 1. Department of Oncology Surgery, Qinghai Provincial People's Hospital, Xi-ning 810001, China;
    2. Department of Oncology, Qinghai Provincial People's Hospital, Xi-ning 810001, China;
    3. Department of Gastroenterology, Qinghai Provincial People's Hospital, Xi-ning 810001, China
  • Received:2019-04-08 Published:2020-04-07

摘要: 目的:探讨LincRNA-p21敲减对胃癌细胞增殖、迁移与侵袭的影响,并阐明其作用机制。方法:采用实时荧光定量聚合酶链反应(RT-qPCR)法检测胃癌组织、癌旁组织以及3种胃癌细胞(MGC-803、MKN-45和SGC-790)和正常胃黏膜上皮细胞GES-1中LincRNA-p21mRNA表达水平。以MGC-803细胞作为研究对象,实验分为sh-NC组、sh-LincRNA-p21组和AG490+sh-LincRNA-p21组。sh-NC组MGC-803细胞采用慢病毒感染sh-NC;sh-LincRNA-p21组MGC-803细胞采用慢病毒感染sh-LincRNA-p21;AG490+sh-LincRNA-p21组MGC-803细胞采用慢病毒感染sh-LincRNA-p21后,再采用10 μg·L-1 AG490处理细胞。采用5-乙炔基-2'-脱氧尿苷(EdU)掺入法检测各组MGC-803细胞EdU掺入百分比,CCK-8法检测各组MGC-803细胞活性,Transwell法检测各组MGC-803细胞迁移数和侵袭数,Western blotting法检测sh-NC组和sh-LincRNA-p21组MGC-803细胞中p-JAK1、p-STAT3和p-STAT5蛋白表达水平。将sh-NC组和sh-LincRNA-p21组MGC-803细胞移植入BALB/c裸鼠颈部皮下成瘤,检测瘤体体积和质量。结果:与癌旁组织比较,胃癌组织中LincRNA-p21mRNA表达水平明显降低(P<0.01);与GES-1细胞比较,MGC-803、MKN-45和SGC-790细胞中LincRNA-p21表达水平均明显降低(P<0.01)。与sh-NC组比较,sh-LincRNA-p21组MGC-803细胞EdU掺入百分比、细胞活性、细胞迁移数和侵袭数、p-JAK1、p-STAT3和p-STAT5蛋白表达水平均明显升高(P<0.05或P<0.01);与sh-LincRNA-p21组小鼠比较,AG490+sh-LincRNA-p21组MGC-803细胞活性、细胞迁移数和侵袭数均明显降低(P<0.05或P<0.01)。裸鼠成瘤实验,与sh-NC组比较,sh-LincRNA-p21组小鼠瘤体体积和质量均明显增加(P<0.05或P<0.01)。结论:敲减LincRNA-p21可明显促进胃癌细胞的生长与转移,且该促进作用可能与其促进JAK-STAT信号通路活性有关。

关键词: LincRNA-p21, 胃肿瘤, JAK-STAT信号通路, 细胞增殖, 细胞迁移, 细胞侵袭

Abstract: Objective: To investigate the effects of LincRNA-p21 knockdown on the proliferation, migration and invasion of gastric cancer cells, and to elucidate their mechanisms. Methods: The expression levels of LincRNA-p21 mRNA in the gastric cancer tissue, adjacent tissue, three types of gastric cancer cells (MGC-803, MKN-45 and SGC-790 cells) and normal gastric mucosal epithelial cells GES-1 were detected by real-time quantitative polymerase chain reaction (RT-qPCR). The MGC-803 cells were used as the subjects and were divided into sh-NC group, sh-LincRNA-p21 group and AG490+sh-LincRNA-p21 group. The MGC-803 cells in sh-NC group were infected with sh-NC by lentivirus; the MGC-803 cells in sh-LincRNA-p21 group were infected with sh-LincRNA-p21 by lentivirus; the MGC-803 cells in AG490+sh-LincRNA-p21 group were infected with sh-LincRNA-p21 by lentivirus, and then treated with 10 μg·L-1 AG490. The percentages of 5-ethynyl-2'-deoxyuridine (EdU) incorporation of the MGC-803 cells in various groups were detected by EdU incorporation assay. The viabilities of MGC-803 cells in various groups were measured by CCK-8 assay. The number of invasion and migration of MGC-803 cells in various groups was detected by Transwell assay. The expression levels of p-JAK1, p-STAT3 and p-STAT5 proeteins in the MGC-803 cells in sh-NC group and sh-LincRNA-p21 group were detected by Western blotting method. The MGC-803 cells from sh-NC group and sh-LincRNA-p21 group were subcutaneouly transplanted into the neck of the BALB/c nude mice, then the volumes and weights of the tumors of the mice were measured. Results: Compared with adjacent tissue, the expression level of LincRNA-p21 mRNA in gastric cancer tissue was significantly decreased (P<0.01); compared with the GES-1 cells, the expression levels of LincRNA-p21 mRNA in the MGC-803, MKN-45 and SGC-790 cells were significantly decreased (P<0.01).Compared with sh-NC group, the percentage of EdU incorporation, the cell viability, the number of migration and invasion cells, the expression levels of p-JAK1, p-STAT3 and p-STAT5 proteins in the MGC-803 cells in sh-LincRNA-p21 group were significantly increased (P<0.05 or P<0.01).Compared with sh-LincRNA-p21 group, the cell viability and the number of migration and invasion of the MGC-803 cells in AG490+sh-LincRNA-p21 group were significantly decreased (P<0.05 or P<0.01).The results of the tumorigenesis of nude mice showed that compared with sh-NC group, the volume and weight of the tumor of the mice in sh-LincRNA-p21 group were significantly increased (P<0.05 or P<0.01). Conclusion: LincRNA-p21 knockdown can significantly promote the growth and metastasis of gastric cancer cells, and this promotion may be related to promoting the activity of JAK-STAT signaling pathway.

Key words: LincRNA-p21, stomach neoplasms, JAK-STAT signaling pathway, cell proliferation, cell migration, cell invasion

中图分类号: 

  • R735.2