摘要
目的探讨HOX转录反义RNA (HOTAIR)在胃癌发病中的作用机制.方法选取40例胃癌患者,其中男29例,女21例,平均年龄(51.7±10.8)岁;Ⅰ期20例,Ⅱ期10例,Ⅲ期6例,Ⅳ期4例;高分化9例,中分化12例,低分化19例.收集所有患者的癌变组织和癌旁组织,实时定量反转录聚合酶链反应(RT-qPCR)法检测两种组织中长链非编码RNA(lncRNA) HOTAIR的表达水平,并对HOTAIR表达与患者临床特征的相关性进行分析;设计合成lncRNA HOTAIR小干扰RNA(siRNA),在原代SGC-7901胃癌细胞株建立沉默模型.检测原代SGC-7901胃癌细胞中lncRNAHOTAIR的表达,流式细胞周期和细胞凋亡检测研究lncRNA HOTAIR对癌细胞增殖凋亡的影响;结合蛋白质印迹法(Western blot)和RT-qPCR技术研究其分子调控机制.应用SPSS 19.0统计软件分析.根据不同的数据类型,组间差异采用χ2检验或t检验分析,相关性采用皮尔森(Pearson's)相关试验评价.结果癌组织中lncRNA HOTAIR的表达量为4.367±0.352,高于癌旁组织的0.984±0.782(P<0.05),癌组织中lncRNA HOTAIR与肿瘤分化程度以及肿瘤分期呈正相关(r=0.347、0.623,P<0.05).lncRNA HOTAIR siRNA转染组与未转染组比较,G1期细胞数升高(78.64土6.91比58.34±6.13,t=3.806,P<O.05),S期细胞数降低(11.93±2.54比31.54±3.01,t =8.624,P<0.05),G2期细胞数量没有明显的变化(9.43±1.47比10.12士2.11,t =0.645,P>0.05).lncRNA HOTAIR siRNA转染处理细胞48 h后,lncRNA HOTAIR siRNA转染组早期凋亡细胞[(22.69±3.21)%比(10.97±1.52)%]、晚期凋亡细胞[(30.41±3.97)%比(7.40±1.71)%]、坏死细胞[(5.53±0.74)%比(3.44±0.42)%]的比率均明显高于未转染组细胞(=5.715、9.220、4.254,P<0.05).转染处理后细胞中Wnt3a蛋白相对表达量明显低于未转染组水平(0.39±0.07比0.92±0.13,t=6.217,P<0.05).结论 lncRNA HOTAIR在胃癌中表达上调,而抑制HOTAIR表达可以通过下调WntWnt3a蛋白表达阻遏Wnt/β-连环蛋白(β-catenin)信号通路的激活进而促进胃癌细胞凋亡.
Objective Investigate the mechanism of HOX antisense intergenic RNA (HOTAIR) in the pathogenesis of gastric cancer. Methods 40 patients with gastric cancer were selected, there were 29 males and 21 females with an average age of (51.7±10.8) years;Ⅰ period 20 cases,Ⅱ period 10 cases,Ⅲ period 6 cases,Ⅳ period 4 cases;9 cases of high differentiation, 12 cases of middle differentiation and 19 cases of low differentiation. The expression levels of long non-coding RNA (lncRNA) HOTAIR in cancer tissues and adjacent tissues of gastric cancer patients were detected by real-time quantitative reverse transcriptase-polymerase chain reaction (RT-qPCR), andanalyzed the correlation between the expression of HOTAIR and the clinical characteristics of gastric cancer patients. lncRNA HOTAIR small interfering RNA (siRNA) were designed and synthesized, and the silencing model was established in the primary SGC-7901 gastric cancer cell line. The expression of lncRNA HOTAIR in primary SGC-7901 gastric cancer cells was detected, add the effects of lncRNA HOTAIR on the proliferation and apoptosis of cancer cellsanalyzed. The molecular regulatory mechanism was studied by Western blotting and RT-qPCR. Results The expression of lncRNA HOTAIR in cancer tissues was (4.367±0.352), higher than that in adjacent tissues (0.984±0.782)(P<0.05). lncRNA HOTAIR in cancer tissues was positively correlated with tumor differentiation degree and tumor staging (r=0.347, r=0.623, P<0.05). Compared with the untransfected group, the number of cells in G1 phase (78.64±6.91 vs. 58.34±6.13, t=3.806, P<0.05) and S phase (11.93 ±2.54 vs. 31.54±3.01, t=8.624, P<0.05)in the lncRNA HOTAIR siRNA transfection group were increased, and there was no significant change in the number of cells in the G2 phase (9.43±1.47 vs. 10.12±2.11, t=0.645, P>0.05). After 48 h of lncRNA HOTAIR siRNA transfection, the ratio of early apoptotic cells [(22.69±3.21)% vs.(10.97±1.52)%], late apoptotic cells [(30.41±3.97)% vs.(7.40±1.71)%] and necrotic cells [(5.53±0.74)
作者
徐勇超
黄涛
唐礼恭
李星
任莹坤
Xu Yongchao;Huang Tao;Tang Ligong;Li Xing;Ren Yingkun(Department of General Surgery,Henan Cancer Hospital,Affiliated Tumor Hospital of Zhengzhou University,Zhengzhou 450008,China;Department of Hepatobiliary and Pancreatic Surgery,Henan Cancer Hospital Affiliated to Zhengzhou University,Zhengzhou 450008,China)
出处
《中华实验外科杂志》
CAS
CSCD
北大核心
2019年第10期1844-1846,共3页
Chinese Journal of Experimental Surgery
