LncRNA SNHG16通过调控miR-195-5p/MYB促进胃癌细胞增殖及迁移  

The mechanism of long non-coding RNA SNHG16 regulating the expression of miR-195-5p to promote the proliferation and migration of gastric cancer cells

作　　者：康娟 贺娇 任伟宏[2] Kang Juan;He Jiao;Ren Weihong(Henan University of Chinese Medicine,Zhengzhou 450046;The First Affiliated Hospital of Henan University of Chinese Medicine,Zhengzhou 450000)

机构地区：[1]河南中医药大学第一临床医学院,郑州450046 [2]河南中医药大学第一附属医院检验科,郑州450000

出　　处：《安徽医科大学学报》2023年第9期1564-1571,1579,共9页Acta Universitatis Medicinalis Anhui

基　　金：国家自然科学基金项目(编号:82174146);河南省科技攻关项目(编号:212102310639)。

摘　　要：目的探讨lncRNA SNHG16(SNHG16)对胃癌细胞增殖、迁移的影响及其分子调控机制。方法基于在线数据库检索SNHG16在胃癌组织中的表达情况并筛选SNHG16的下游靶基因。通过生物信息学方法分析、双荧光素酶报告基因实验验证SNHG16与miR-195-5p间相互作用关系。实时荧光定量PCR(qRT-PCR)检测SNHG16、miR-195-5p和MYB在胃癌细胞(HGC-27、MKN-28)中的表达情况;敲低SNHG16检测miR-195-5p或MYB表达;过表达miR-195-5p检测MYB表达;蛋白质印迹分析(Western blot)检测各组中MYB的蛋白表达水平。敲低SNHG16或过表达miR-195-5p,通过细胞增殖及划痕实验分别检测胃癌细胞(HGC-27、MKN-28)增殖及迁移能力。结果LncRNA SNHG16在胃癌组织及胃癌细胞(HGC-27、MKN-28、MKN-45、NCI-N87)中表达升高。双荧光素酶报告基因实验结果显示,将psiCHECK2-SNHG16-WT和miR-195-5p mimic同时转染进HGC-27中,显著抑制了HGC-27细胞的荧光素酶活性。qRT-PCR及WB实验结果显示:敲低HGC-27、MKN-28细胞中SNHG16可上调miR-195-5p并抑制MYB在转录及翻译水平的表达;过表达HGC-27、MKN-28细胞中miR-195-5p可抑制MYB表达。CCK8增殖实验及细胞划痕实验结果表明:敲低SNHG16或过表达miR-195-5p均可抑制HGC-27、MKN-28细胞的增殖和迁移。结论LncRNA SNHG16可通过miR-195-5p调节MYB在胃癌细胞中的表达,SNHG16高表达可促进胃癌细胞增殖和迁移。Objective To investigate the effect of lncRNA SNHG16(SNHG16)on the proliferation and migration of gastric cancer cells and its molecular regulatory mechanism.Methods The expression of SNHG16 in gastric cancer tissue was retrieved based on online database and the downstream target gene of SNHG16 was screened.The interaction between SNHG16 and miR-195-5p was verified by bioinformatics analysis and double luciferase reporter gene experiment.The expression of SNHG16,miR-195-5p and MYB in gastric cancer cells(HGC-27,MKN-28)was detected by real-time fluorescent quantitative PCR(qRT-PCR);SNHG16 was knocked down to detect the expression of miR-195-5p or MYB;The expression of MYB was detected by overexpression of miR-195-5p;Western blot analysis was used to detect the protein expression level of MYB in each group.Knock down SNHG16 or overexpress miR-195-5p,and the proliferation and migration ability of gastric cancer cells(HGC-27,MKN-28)were detected through cell proliferation and scratch test.Results The expression of LncRNA SNHG16 increased in gastric cancer tissues and gastric cancer cells(HGC-27,MKN-28,MKN-45,NCI-N87).The results of double luciferase reporter gene experiment showed that psiCHECK2-SNHG16-WT and miR-195-5p mimic were transfected into HGC-27 at the same time,significantly inhibiting the luciferase activity of HGC-27 cells.The results of qRT PCR and WB experiments showed that knocking down SNHG16 in HGC-27 and MKN-28 cells upregulated miR-195-5p and inhibited the expression of MYB at the transcription and translation levels;Overexpression of miR-195-5p in HGC-27 and MKN-28 cells inhibited MYB expression.The results of CCK8 proliferation test and cell scratch test showed that knocking down SNHG16 or overexpressing miR-195-5p could inhibit the proliferation and migration of HGC-27 and MKN-28 cells.Conclusion LncRNA SNHG16 can regulate the expression of MYB in gastric cancer cells through miR-195-5p,and the high expression of SNHG16 can promote the proliferation and migration of gastric cancer cells.

关 键 词：lncRNA SNHG16 miR-195-5p/MYB 胃癌细胞 增殖 迁移 

分 类 号：R735.2[医药卫生—肿瘤]