Effect of miRNA interference to annexinA3 gene on growth of human gallbladder cancer cells
10.3760/cma.j.issn.1007-8118.2010.11.018
- VernacularTitle:RNA干扰人膜联蛋白A3对胆囊癌细胞增殖的影响
- Author:
Yi TAN
;
Haiping MENG
;
Fengmeng TENG
;
Xingsheng JIA
;
Haorong WU
- Publication Type:Journal Article
- Keywords:
Gallbladder neoplasms;
Interfering RNA;
AnnexinA3
- From:
Chinese Journal of Hepatobiliary Surgery
2010;16(11):853-856
- CountryChina
- Language:Chinese
-
Abstract:
Objective To construct a recombinant interfering RNA(miRNA)plasmid vector targeting annexinA3 and observe its impact on the growth and apoptosis of human gallbladder cancer cell line SGC-996.Methods Four small fragments of miRNA and one negative control were sequenced and cloned into vector pcDNATM 6.2-GW/EmGFPmiR.The recombinant plasmids were then transfected into gallbladder cancer cell line SGC-996 by positive liposomal transfection.The transcription and translation level of annexinA3 expression were detected by RT-PCR and Western Blot.Meanwhile, the proliferation and apoptosis of transfected tumor cells were determined by MTT and flow cytometry.Results The recombinant plasmids containing annexinA3 miRNA were successfully constructed.AnnexinA3 expression was significantly knocked down after miRNA plasmid transfection.Transfection of annexinA3 gene miRNA could effectively inhibit the proliferation of gallbladder cancer cell and promote apoptosis.Conclusion AnnexinA3 gene silenced by miRNA can induce the apoptosis of human gallbladder cancer cell line SGC-996.The process might serve as a potential approach for cancer gene therapy.
