[1]王密斯,张荣花,王琳,等.miR-185对肝癌细胞生长增殖及侵袭迁移的影响[J].陆军军医大学学报(原第三军医大学学报),2016,38(17):1918-1923.
 Wang Misi,Zhang Ronghua,Wang Lin,et al.Effect of miR-185 on proliferation, invasion and migration of hepatocellular carcinoma cells[J].J Amry Med Univ (J Third Mil Med Univ),2016,38(17):1918-1923.
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miR-185对肝癌细胞生长增殖及侵袭迁移的影响(/HTML )
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陆军军医大学学报(原第三军医大学学报)[ISSN:1000-5404/CN:51-1095/R]

卷:
38卷
期数:
2016年第17期
页码:
1918-1923
栏目:
基础医学
出版日期:
2016-09-15

文章信息/Info

Title:
Effect of miR-185 on proliferation, invasion and migration of hepatocellular carcinoma cells
作者:
王密斯张荣花王琳孔艺璇冯雪研刘志勇甄永占章广玲
华北理工大学:基础医学院病原生物学与免疫学教研室,附属医院胸心外科,基础医学院组织学与胚胎学教研室,基础医学院河北省慢性疾病重点实验室,唐山市慢性病临床基础研究重点实验室
Author(s):
Wang Misi Zhang Ronghua Wang Lin Kong Yixuan Feng Xueyan Liu Zhiyong Zhen Yongzhan Zhang Guangling

Department of Pathogen Biology and Immunology, Department of Histology and Embryology,Hebei Key Laboratory for Chronic Diseases, Tangshan Key Laboratory for Preclinical and Basic Research on Chronic Diseases, School of Basic Medical Sciences; Department of Chest Cardiac Surgery, Affiliated Hospital, North China University of Science and Technology, Tangshan, Hebei Province, 063000, China

关键词:
miR-185肝癌细胞细胞增殖肿瘤浸润细胞运动
Keywords:
miR-185 hepatoma carcinoma cells proliferation invasion migration
分类号:
R394.2; R730.23; R735.7
文献标志码:
A
摘要:

目的     探讨miR-185对人肝癌细胞HepG2和SMMC-7721生长增殖、侵袭迁移能力的影响。   方法      构建miR-185过表达载体、合成miR-185反义寡聚核苷酸(antisense oligonucleotide, ASO),经脂质体转染肝癌细胞HepG2和SMMC-7721。利用CCK-8和Transwell小室实验检测细胞的增殖、侵袭和迁移能力;生物信息学数据库预测结合基因芯片结果确定miR-185的候选靶基因NR1D1;利用实时荧光定量PCR技术和Western blot检测miR-185对细胞中NR1D1基因表达水平的影响;荧光报告载体实验验证miR-185对靶基因的调控作用。    结果    miR-185在人肝癌细胞中的mRNA表达水平显著低于正常肝细胞(P<0.01);过表达miR185使人肝癌细胞的增殖、侵袭和迁移能力降低,下调肝癌细胞中NR1D1基因的mRNA和蛋白的表达水平(P<0.01);荧光报告载体实验证明miR-185可以通过作用于NR1D1 mRNA的3′端非翻译区(untranslated region, UTR)下调其表达水平(P<0.05)。  结论     miR-185能抑制肝癌细胞的增殖、侵袭迁移能力,NR1D1是miR-185的直接靶基因。

Abstract:

Objective    To determine the effect of miR-185 on the proliferation, invasion and migration in human hepatic carcinoma cell lines HepG2 and SMMC-7721. Methods     After miR-185 expression vector was constructed and miR-185 antisense oligonucleotide (ASO) was synthesized, they were used to transfect the HepG2 and SMMC-7721 cells. Cell counting kit-8 assay and Transwell chamber test were employed to detect cell proliferation, invasion and migration. Bioinformatics database combined with gene chip results indicated that NR1D1 may be the candidate target gene of miR-185. So real-time quantitative PCR and Western blotting were adopted to detect the mRNA and protein levels of NR1D1. Fluorescent report carrier experiment was adopted to detect the regulation function of miR-185 with its target gene.   Results    The mRNA level of miR-185 was significantly lower in human hepatic carcinoma cells than that of normal liver cells (P<0.05). Overexpression of miR-185 resulted in the decreases in proliferation, invasion and migration of the hepatic carcinoma cells, and the down-regulation of NR1D1 at mRNA and protein levels (P<0.01). Fluorescent report carrier experiment proved that miR-185 acted on the 3′ untranslated region of NR1D1 mRNA to down-regulate its expression level (P<0.05).  Conclusion    Overexpression of miR-185 may inhibit the proliferation, invasion and migration in hepatic carcinoma cells. NR1D1 may be a target gene of miR-185.

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更新日期/Last Update: 2016-09-06