The Study Of Up-regulation Of P21^WAF1/CIP1 By RNAa In Gallbladder Cancer Cells In Vitro

Objective:To observe the effects of p21WAF1/CIP1activation by RNAa on proliferation,invasion and migration of human gallbladder cancer cell.To investigate the possibility of gallbladder cancer gene therapy and molecular mechanism.To provide a theoretical foundation and experimental basis for gene therapy of gallbladder cancerMethods:We transfected dsRNA into gallbladder cancer cell line by using LipofectamineTM2000.We divided into three groups:one is blank group(only liposomes),another is negative group(the control dsRNA is non-homologous to the whole human genome),the other is experiment group(dsp21-322is targetedthe p21gene promoter at position-322)1.Expression of mRNA and protein was evaluated by semi quantitative RT-PCR and Western blot.2.The gallbladder cancer cell proliferation was detected by MTT assay.3.The migrative and invasive ability of GBS-SD cells were determined by Transwell chamber assay.Results:1.A dsRNA targeting the p21WAF1/CIP1gene promoter at position-322relative to the transcription start site was transfected into gallbladder cancer cells. At72h after the GBS-SD cells were transected with dsRNA,the results of semi quantitative RT-PCR and Western blot revealed that the expression of p21WAF1/CIP1mRNA and protein were up-regulated.2.Compared with the blank group and negative group,the proliferative activity of the cells was reduced significantly3.Compared with the blank group and negative group,the ability of migration and invasion of the cells were decreased.Conclusions:1.At72h after the GBS-SD cells were transected with dsRNA,dsp21-322can effectively up-regulate the expression of p21WAF1/CIP1gene mRNA and protein.2.After dsp21-322was transfected into GBC-SD cells,it has an substantially inhibitor effect on cell proliferation and can inhibit the invasion and metastasis of GBC-SD cells in vitro. 3.RNA activation provides a new method to the gene therapy of gallbladder cancer.