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  • 吴为明,李先鹏,郭世伟,金海波,卿艳平.miR-181a调控结肠癌细胞生长的作用和机制研究[J].第二军医大学学报,2014,35(5):574-576    [点击复制]
  • WU Wei-ming,LI Xian-peng,GUO Shi-wei,JIN Hai-bo,QING Yan-ping.Role of miR-181a in regulating colon cancer cell growth and the related mechanism[J].Acad J Sec Mil Med Univ,2014,35(5):574-576   [点击复制]
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miR-181a调控结肠癌细胞生长的作用和机制研究
吴为明1,李先鹏1,郭世伟2,金海波1,卿艳平1*
0
(1. 宁波大学医学院附属医院肛肠科,宁波 315020;
2. 第二军医大学长海医院普通外科,上海 200433
*通信作者)
摘要:
目的 确定miR-181a对结肠癌细胞生长的作用以及探讨其机制。方法 常规培养结肠癌细胞系TCHu102,利用脂质体转染miR-181a抑制剂/模拟物的方法干预细胞内源性miR-181a的水平。MTT法测定结肠癌细胞的生长活力。生物信息学预测的方法筛选miR-181a的可能靶基因,双荧光素酶报告基因确定miR-181a与靶基因的作用关系。实时定量PCR的方法确定基因表达水平,western blot的方法检测蛋白表达水平。结果 敲低结肠癌细胞TCHu102中miR-181a的水平明显抑制细胞生长活力,为对照组的28.9% (P<0.01)。生物信息学预测发现PRKCD基因可能是miR-181a的靶基因,同时miR-181a能显著抑制含有PRKCD基因3’UTR的荧光素酶活性,在结肠癌细胞中过表达miR-181a后PRKCD的蛋白水平明显降低。结论 miR-181a通过抑制PRKCD的表达调控结肠癌细胞的生长。
关键词:  miR-181a  结肠肿瘤  细胞增殖  PRKCD  肿瘤基因表达调控
DOI:10.3724/SP.J.1008.2014.00574
投稿时间:2013-11-15修订日期:2014-04-22
基金项目:中国博士后科学基金(2013M542436).
Role of miR-181a in regulating colon cancer cell growth and the related mechanism
WU Wei-ming1,LI Xian-peng1,GUO Shi-wei2,JIN Hai-bo1,QING Yan-ping1*
(1. Department of Anorectal, the Affiliated Hospital of Ningbo University School of Medicine, Ningbo 315020, Zhejiang, China;
2. Department of General Surgery, Changhai Hospital, Second Military Medical University, Shanghai 200433, China
*Corresponding authors.)
Abstract:
Objective To determine the role and mechanism of miR-181a on growth of colon cancer cells. Methods The colon cancer cell lines TCHu102 was cultured routinely, and the endogenous miR-181a was interferedreduced by liposomal transfection of inhibitor or mimic. The growth activity of colon cancer cell was determined by MTT assay. The potential targeted genes of miR-181a were predicted by bioinformatics, and the relationship between miR-181a and PRKCD were identified by dual-luciferase reporter assay. The expression level of gene were detected by real-time PCR and western blot respectively. Result The growth activity of colon cancer cell with low miR-181a level was significantly inhibited, with 28.9% of the control group (P<0.01). PRKCD was predicted as the potential targeted gene of miR-181a, and miR-181a could suppress the luciferase activity containing PRKCD 3’UTR. Moreover, the protein levels of PRKCD was significantly lower in colon cancer cells overexpressing miR-181a. Conclusion miR-181a regulated the growth of colon cancer cells by inhibition of PRKCD expression.
Key words:  miR-181a  colonic neoplasms  cell proliferation  PRKCD  neoplastic gene expressionregulation