Effect of miRNA interference to annexinA3 gene on growth of human gallbladder cancer cells / 中华肝胆外科杂志

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.