摘要
目的观察mi RNA-381对胃癌细胞增殖和侵袭的影响,并探讨其作用机制。方法应用实时荧光定量聚合酶链反应(q RT-PCR)检测胃癌细胞系AGS、MGC-803、Hs746T及BSG823和正常胃黏膜上皮细胞系RGM-1中mi RNA-381的表达;将AGS细胞系分成两组,阴性对照组和mi RNA-381模拟物组,采用Lipofectamine TM 2000分别转染mi RNA-381 scramble及mimics,采用CCK-8、Transwell实验测定两组细胞增殖和侵袭的影响,Western blot检测两组细胞肝受体类似物1(LRH-1)和扭曲相关蛋白1(Twist1)表达水平。结果胃癌细胞系AGS、MGC-803、Hs746T及BSG823中mi RNA-381相对表达量较正常胃黏膜上皮细胞系RGM-1为低。mi RNA-381模拟物组在转染后72和96 h OD 450 nm值低于阴性对照组(P<0.05);阴性对照组侵袭细胞数为(79.6±7.2)个,mi RNA-381模拟物组侵袭细胞数为(31.70±4.2)个,mi RNA-381模拟物组侵袭细胞数少于阴性对照组(P<0.01);mi RNA-381模拟物组LRH-1蛋白相对表达量为(0.39±0.04),阴性对照组为1.0,mi RNA-381模拟物组LRH-1蛋白相对表达量低于阴性对照组(P<0.01);mi RNA-381模拟物组Twist1蛋白相对表达量为(0.51±0.05),阴性对照组为1.0,mi RNA-381模拟物组Twist 1蛋白相对表达量低于阴性对照组(P<0.01)。结论 mi RNA-381低表达于胃癌细胞系,mi RNA-381过表达可抑制胃癌细胞增殖和侵袭能力,其机制可能与下调LRH-1和Twist1的表达有关。
Objective To investigate the role of miR-381 on proliferation and invasion of gastric carcinoma cells and its potential mechanism. Methods The expression of miR-381 in normal and gastric cell lines were determined by qRT-PCR. The AGS cell line was divided into two groups including negative control group which was transfected with miR-381 vehicle, and miR-381 mimics group which was transfected with miR-381 mimics through Lipofectamine 2000. Proliferation and invasion rate were evaluated with CCK8 and Transwell assay, respectively. LRH-1 and Twist1 protein levels were measured by Western blot. Results Substantially reduction of miR-381 was found in gastric cancer cell line when compared with normal gastric cell line （P 〈 0.05）. The number of invasive cell in miR-381 mimics group was decreased signifcantly compared with negative control group [（31.70 ± 4.2） vs （79.6 ± 7.2）, P 〈 0.01]. Expression levels of LRH-1 and Twist1 signifcantly dropped in miR-381 mimics group in comparison with those in negative control group [（0.39 ± 0.04） vs （1.0 ± 0.0）, （0.51 ± 0.05） vs （1.0 ± 0.05）, P 〈 0.01, respectively]. Conclusions Low expression of miR-38 is identified in gastric cell line. Up-regulated expression of miR-381 contributes to inhibition of cellular proliferation and invasion activity through LRH-1and Twist signal pathway.
出处
《中国现代医学杂志》
CAS
2018年第6期9-14,共6页
China Journal of Modern Medicine
基金
湖北省卫生厅资助课题(No:WJ2017M177)
