摘要
背景长链非编码RNA长基因间非蛋白编码RNA 996(long intergenic non-protein coding RNA 996,LINC00996)在多种肿瘤中发挥促癌或抑癌作用,而其在胃癌中的表达和作用尚不清楚.目的探索LINC00996在胃癌组织和细胞系中的表达,并探讨其对胃癌细胞生物学行为的作用及机制.方法用生物信息学法分析胃癌中LINC00996的表达,及其对胃癌患者总生存期的影响.用RT-qPCR检测胃癌及癌旁组织、胃癌及正常胃上皮细胞系中LINC00996表达.胃癌细胞(SGC7901、NCI-N87)转染针对LINC00996的小干扰RNA(si-LINC00996)和细胞周期蛋白依赖性激酶抑制蛋白2A(cyclin-dependent kinase inhibitor 2A,CDKN2A)的小干扰RNA(si-CDKN2A)后,用细胞计数试剂盒-8法、EDU染色法、流式细胞术法、划痕法和Transwell法分别检测细胞增殖、细胞周期、凋亡、迁移与侵袭;Western blot法检测细胞中CDKN2A、周期蛋白D1、B淋巴细胞瘤-2(B-cell lymphoma-2,Bcl-2)、Bcl-2相关X蛋白(Bcl-2 associated X protein,Bax)和劈开的半胱胺酸蛋白酶-3(Cleaved cysteinyl aspartate-specific proteinase-3,Cleaved caspase-3)表达.结果LINC00996在胃癌组织和细胞中表达较癌旁组织和胃正常细胞系显著升高(P<0.05),Kaplan Meier Plotter数据库显示LINC00996高表达组的胃癌患者的总生存期较低表达组显著缩短(P<0.05).敲降LINC00996能抑制胃癌细胞增殖、细胞周期运行、迁移、侵袭(P<0.05),促进细胞凋亡(P<0.05);降低cyclin D1和Bcl-2表达(P<0.05);升高CDKN2A、Bax和Cleaved caspase-3表达(P<0.05).敲降CDKN2A能部分逆转敲降LINC00996对胃癌细胞的作用(P<0.05).结论敲降LINC00996能通过上调CDKN2A诱导胃癌细胞凋亡,抑制其增殖、迁移与侵袭.
BACKGROUND The expression and function of the long chain noncoding RNA(lncRNA)long intergenic non-protein coding RNA 996(LINC00996)in gastric cancer remain elusive,despite its established pro-or anti-tumor roles across various tumor types.AIM To investigate the expression of LINC00996 in gastric cancer tissues and cell lines,and to explore its role and underlying mechanism in the biological behavior of gastric cancer cells.METHODS Bioinformatics was used to analyze the expression of LINC00996 in gastric cancer and its effect on the overall survival of patients with gastric cancer.The expression of LINC00996 in gastric cancer and adjacent tissues as well as gastric cancer and normal gastric epithelial cell lines was detected by RT-qPCR.Gastric cancer cells(SGC7901 and NCI-N87)were transfected with small interfering RNA targeting LINC00996(si-LINC00996)and cyclin-dependent kinase inhibitor 2A(CDKN2A)(si-CDKN2A),and cell proliferation,cell cycle progression,apoptosis,migration,and invasion were detected by cell counting kit-8,EDU staining,flow cytometry,scratch assay,and Transwell assay,respectively.The expression of CDKN2A,cyclin D1,B-cell lymphoma-2(Bcl-2),Bcl-2 associated X protein(Bax),and cleaved cysteinyl aspartate-specific proteinase-3(caspase-3)was detected by Western blot.RESULTS The expression of LINC00996 in gastric cancer tissues and cells was significantly higher than that in adjacent tissues and normal gastric cell lines(P<0.05).Survival analysis based on the Kaplan-Meier Plotter database showed that the overall survival of gastric cancer patients with high expression of LINC00996 was significantly shorter than that of patients with low expression(P<0.05).Knockdown of LINC00996 inhibited the proliferation,cell cycle progression,migration,and invasion of gastric cancer cells(P<0.05),promoted apoptosis(P<0.05),decreased the expression of cyclin D1 and Bcl-2(P<0.05),and increased the expression of CDKN2A,Bax,and cleaved caspase-3(P<0.05).Knockdown of CDKN2A partially reversed the effect of LINC00996 knockdown on gastric cancer cells(P<0.05).CONCLUSION Knockdown of LINC00996 can induce apoptosis of gastric cancer cells and inhibit their proliferation,migration,and invasion by up-regulating CDKN2A expression.
作者
程国雄
刘明
陈正伟
叶巧萍
Guo-Xiong Cheng;Ming Liu;Zheng-Wei Chen;Qiao-Ping Ye(Department of Gastroenterology,Lishui People’s Hospital,Lishui 323000,Zhejiang Province,China)
出处
《世界华人消化杂志》
CAS
2024年第4期302-312,共11页
World Chinese Journal of Digestology
基金
丽水市科技计划项目,No.2020SJZC037.
作者简介
程国雄,本科,副主任医师,研究方向为胃肠道肿瘤外科综合治疗的临床和基础研究;通讯作者:程国雄,本科,副主任医师,323000,浙江省丽水市丽阳街1188号,丽水市人民医院胃肠外科.jiemengqn52403@163.com。
