RNA激活上调胆囊癌细胞p21WAF1CIP1基因表达的体外实验研究.pdfVIP

Abstract WAF1/CIP1 The Study of Up-regulation of p21 by RNAa in Gallbladder Cancer Cells in Vitro Objective:To observe the effects of p21WAF1/CIP1 activation by RNAa on proliferation,invasion and migration of human gallbladder cancer cell. Methods:A dsRNA targeting the p21WAF1/CIP1 gene promoter at position-322 relative to the transcription start site was transfected into gallbladder cancer cells.Expression of mRNA and protein was evaluated by semiquantitative RT-PCR and Western blot.Cell proliferation was detected by MTT assay.The migrative and invasive ability of GBS-SD cells were determined by Transwell chamber assay. Results:A dsRNA targeting the p21WAF1/CIP1 gene promoter at position-322 relative to the transcription start site was transfected into gallbladder cancer cells.At 72 h after the GBS-SD cells were transected with dsRNA,the results of semiquantitative RT-PCR and Western blot revealed that the expression of p21WAF1/CIP1 mRNA and protein were up-regulated.Compared with the blank group and negative group,the proliferative activity of the cells was reduced significantly and the ability of migration and invasion of the cells were decreased. Conclusions:1.At 72 h after the GBS-SD cells were transected with dsRNA,dsp21-322 can effectively up-regulate the expression of p21WAF1/CIP1 gene mRNA and protein. 2.After dsp21-322 was 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. Key words:Gallbladder cancer;RNA activation;p21W