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《天津医科大学学报》[ISSN:1006-8147/CN:12-1259/R]

卷:

22卷

期数:

2016年01期

页码:

13-16

栏目:

基础医学

出版日期:

2016-01-20

文章信息/Info

Title:

Experimental research on reducing invasion and metastasis ability of HCT-116 by inhibiting microRNA-21 expression

文章编号:

1006-8147（2016）01-0013-04

作者:

王金淼; 姜 涛; 戚 峰; 刘 彤; 王鹏志

(天津医科大学总医院普通外科,天津300052)

Author(s):

WANG Jin-miao;  JIANG Tao; QI Feng; LIU Tong; WANG Peng-zhi

(Department of General Surgery，General Hospital，Tianjin Medical University，Tianjin 300052，China)

关键词:

结肠癌; 侵袭; 转移; microRNA-21; 质粒

Keywords:

colonic cancer;  invasion;  metastasis;  microRNA-21;  plasmid

分类号:

R735.3+5

DOI:

-

文献标志码:

A

摘要:

目的:探讨抑制人结肠癌细胞HCT-116 microRNA-21表达降低结肠癌细胞侵袭转移能力作用机制。方法:应用表达反义microRNA-21的质粒miRZip21转染HCT-116细胞，检测microRNA-21的变化，TargetScan和PicTar等工具筛选与侵袭转移相关的靶基因，实时定量PCR方法检测转染前后靶基因mRNA的变化，Western Blot检测靶基因蛋白表达变化，平皿划痕和Transwell实验观察抑制microRNA-21与否HCT-116细胞转移和侵袭能力。结果:筛选出4种与侵袭转移相关的microRNA-21靶基因：TIMP-3，RECK，BMPRⅡ和PCDH17；miRZip21质粒转染HCT-116后，micro-RNA 21表达水平下降约40%，靶基因TIMP-3和RECK 的mRNA及靶蛋白表达上升明显，有统计学意义（P <0.05）；平皿划痕和Transwell实验结果表明抑制microRNA-21后，HCT-116细胞转移侵袭能力明显减弱，差异有统计学意义（P <0.05）。结论:抑制microRNA-21的表达能够减弱HCT-116细胞的侵袭和转移能力。

Abstract:

Objective: To explore the effect on invasion and metastasis of HCT-116 colon cancer cell by downregulating the microRNA-21 expression. Methods: After plasmid miRZip21, miRZip^TManti-microRNA-21expression lentivector , was used to transfect HCT-116, the expression of microRNA-21 was assessed. The microRNA-21 target genes associated with invasion and metastasis were predicted by TargetScan and PicTar. The mRNA and protein of taget genes were detected by qRT-PCR and Western blotting respectively. The ability of migration and invasion of HCT-116 after transfection were detected by the Scratch assay and Transwell assay. Results: TIMP-3, RECK, BMPRⅡ and PCDH17 were the gene targets associated with invasion and metastasis; After the cells were transfected with plasmid miRZip21，the microRNA-21 level descended to about 60%，the mRNA and protein level of TIMP-3 and RECK increased significantly (P<0.05); Significant reduction of invasion and migration were observed in anti-microRNA-21-transfected HCT-116 (P <0.05). Conclusion: Inhabiting the microRNA-21 expression can lessen the invasive and metastatic ability of HCT-116 cells.

参考文献/References:

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备注/Memo

备注/Memo:

基金项目 天津市自然科学基金资助项目（08JCYBJC08100）

作者简介 王金淼（1987-），男，硕士在读，研究方向：普通外科；通信作者：戚峰，E-mail：qf@medmail.com.cn。

常用功能

更新日期/Last Update: 2016-01-25