miR-335-5p通过靶向Oct4调控胃癌细胞的增殖

被引：2
|
作者：
程安琪
彭旭东
刘光艺
王子卫
机构：
[1] 重庆医科大学第一附属医院胃肠外科
来源：
重庆医科大学学报 | 2019年 / 44卷 / 03期
关键词：
miR-335-5p; 胃癌; 细胞增殖; Oct4;
D O I：
10.13406/j.cnki.cyxb.001994
中图分类号：
R735.2 [胃肿瘤];
学科分类号：
摘要：
目的:研究微小(mi R)-335-5p对胃癌细胞增殖的影响及其机制。方法:采用荧光定量PCR检测mi R-335-5p在胃癌细胞系SGC-7901、BGC-823、MKN-28、MKN-45和人正常胃黏膜上皮细胞系GES-1中的表达水平,利用mi R-335-5p过表达慢病毒及封闭慢病毒分别感染SGC-7901和BGC-823细胞株,实验分为SGC-7901细胞中mi R-335-5p组(转染过表达mi R-335-5p组)和阴性对照组(空病毒组,NC),BGC-823细胞中antago-mi R-335-5p组(转染封闭mi R-335-5p慢病毒)和阴性对照组(空病毒组,NC)。通过CCK-8实验、平板克隆实验、Ed U实验检测mi R-335-5p对细胞增殖的影响,用生物信息学软件预测mi R-335-5p可能的靶基因,通过Western blot验证mi R-335-5p对靶基因的调控作用并检测AKT及p AKT的表达。结果:mi R-335-5p在胃癌细胞系中表达明显下降。CCK-8实验、平板克隆实验及Ed U实验中过表达mi R-335-5p组光密度值、克隆形成率、Ed U阳性细胞率均低于其阴性对照组,而封闭mi R-335-5p组光密度值、克隆形成率、Ed U阳性细胞率均高于其阴性对照组。同时用生物信息学软件预测了Oct4作为mi R-335-5p的靶基因,Western blot表明过表达mi R-335-5p组Oct4的表达量(0.469±0.054)明显低于阴性对照组(0.670±0.045)(P=0.008),封闭mi R-335-5p组Oct4的表达(0.804±0.046)高于阴性对照组(0.600±0.073)(P=0.015),同时双荧光素酶报告实验证明mi R-335-5p能够靶向Oct4;过表达mi R-335-5p组中p AKT的表达量(0.525±0.046)较阴性对照组(0.847±0.053)有所下降(P=0.001),封闭mi R-335-5p组(0.841±0.069)中p AKT的表达量较阴性对照组(0.563±0.063)有所升高(P=0.007)。结论:mi R-335-5p可以通过靶向Oct4抑制AKT通路从而抑制胃癌细胞的增殖。
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页码：268 / 274
页数：7
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