| 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）. |
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| 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 |
