长链非编码RNA SNHG14通过靶向miR-144-3p调控胃癌细胞增殖和凋亡的体外实验研究  

Long-chain non-coding RNA SNHG14 regulates proliferation and apoptosis of gastric cancer cells by targeting miR-144-3p

作　　者：李昊天 裴效瑞[1] 李洪涛[1] 郝明利[1] Hao-Tian Li;Xiao-Rui Pei;Hong-Tao Li;Ming-Li Hao(General Surgery Department of Tianjin Taida Hospital Tianjin 300457,China)

机构地区：[1]天津市泰达医院普外科

出　　处：《世界华人消化杂志》2019年第21期1304-1312,共9页World Chinese Journal of Digestology

摘　　要：背景长链非编码RNA(long-chain non-coding RNA,LncRNA)与胃癌发生发展密切相关,LncRNA SNHG14在肿瘤发生过程中发挥癌基因作用,但其在胃癌中的作用机制尚未阐明.starBase预测显示miR-144-3p可能是SNHG14的靶基因,但SNHG14是否可通过调控miR-144-3p的表达而参与胃癌发生过程尚未可知.目的探讨LncRNA SNHG14是否可通过靶向miR-144-3p调控胃癌细胞增殖与凋亡.方法实时荧光定量聚合酶链反应检测人胃黏膜上皮正常细胞与胃癌细胞中SNHG14与miR-144-3p的表达情况.体外培养人胃癌MGC-803细胞,随机分为5组:空白(NC)组、阴性对照(si-con)组、SNHG14小干扰RNA(si-SNHG14)组、SNHG14小干扰RNA+miR-144-3p阴性对照(si-SNHG14+anti-miR-con)组、SNHG14小干扰RNA+miR-144-3p特异性寡核苷酸抑制剂(si-SNHG14+anti-miR-144-3p)组.甲基噻唑基四唑(methyl thiazolyl tetrazolium,MTT)法检测各组细胞的增殖情况;流式细胞术检测各组细胞凋亡率;双荧光素酶报告系统验证SNHG14与miR-144-3p的靶向调节关系.蛋白免疫印迹法(Western blot)检测细胞中细胞周期蛋白1(cyclinD1)、B淋巴细胞瘤2(B-cell lymphoma-2,Bcl-2)、p21、Bcl-2相关X蛋白(Bcl-2-associated X protein,Bax)、活化的含半胱氨酸的天冬氨酸蛋白水解酶3(cleaved caspase-3)、磷脂酰肌醇激酶(phosphoinositide 3-kinase,PI3K)/蛋白激酶B(protein kinase B,AKT)信号通路相关蛋白表达.结果SNHG14在胃癌细胞中的表达水平显著高于胃黏膜上皮正常细胞(P<0.05),而miR-144-3p的表达水平显著降低(P<0.05);与NC组、si-con组比较,si-SNHG14组MGC-803细胞OD值显著降低(P<0.05),细胞凋亡率显著增高(P<0.05),cyclinD1、Bcl-2、PI3K的磷酸化(p-PI3K)、AKT的磷酸化(p-AKT)蛋白表达显著降低(P<0.05),p21、Bax、cleaved caspase-3蛋白表达显著升高(P<0.05);双荧光素酶报告基因显示SNHG14可靶向结合miR-144-3p,并可负向调控miR-144-3p的表达与活性;干扰miR-144-3p可部分逆转沉默SNHG14对胃癌细胞增殖、凋BACKGROUND Long-chain non-coding RNAs(lncRNAs)are closely related to the development of gastric cancer.LncRNA SNHG14 play an oncogene role in tumorigenesis,but its mechanism of action in gastric cancer has not been elucidated.Bioinformatics prediction showed that miR-144-3p may be a target gene of SNHG14,but whether SNHG14 participates in the development of gastric cancer by regulating the expression of microRNA-144-3p(miR-144-3p)is unknown.AIM To investigate whether the lncRNA SNHG14 can regulate the proliferation and apoptosis of gastric cancer cells by targeting miR-144-3p.METHODS Real-time quantitative polymerase chain reaction was used to detect the expression of SNHG14 and miR-144-3p in normal human gastric epithelial cells and gastric cancer cells.Human gastric cancer MGC-803 cells were cultured in vitro and randomly divided into five groups:NC group,si-con group,si-SNHG14 group,si-SNHG14+anti-miR-con group,and si-SNHG14+antimiR-144-3p group.MTT assay was used to detect the proliferation of cells in each group.Flow cytometry was used to detect the apoptosis of cells.The dual luciferase reporter system was used to validate whether there is a targeted regulatory relationship between SNHG14 and miR-144-3p.The expression of CyclinD1,Bcl-2,p21,Bcl-2-associated X protein(Bax),and PI3K/AKT signaling pathway-related proteins was detected by Western blot.RESULTS The expression level of SNHG14 in gastric cancer cells was significantly higher than that in normal gastric epithelial cells(P<0.05),while the expression level of miR-144-3p was significantly decreased(P<0.05).Compared with the NC group and si-con group,the proliferation of MGC-803 cells in the si-SNHG14 group was significantly decreased(P<0.05),and the apoptosis rate was significantly increased(P<0.05).The expression of p-AKT,CyclinD1,Bcl-2,and p-PI3K protein was significantly decreased(P<0.05),while the expression of p21,Bax,and cleaved caspase-3 protein was significantly increased(P<0.05).The dual luciferase reporter assay showed that SNHG14 can bind to

关 键 词：LncRNA SNHG14 miR-144-3p 胃癌 增殖 凋亡 

分 类 号：R73[医药卫生—肿瘤]