The Inhibitory Effect of Exogenous Wild-Type p53 Gene on the Cell Growth and Tumorigenicity of Human Gallbladder Cancer Cell Lines / 中国肿瘤生物治疗杂志

ABSTRACT

Objective: To investigate effect of exogenous wild-type p53 gene on the cell growth and tumorigenicity of human gallbladder cancer cell lines. Methods: After identification of the genetic status of p53 gene of GBC-SD cell lines with the immunocytochemistry staining and the direct sequencing technique of PCR products, eukaryotic expressing plasmid pCMV-p53 was introduced by lipofectamine-mediated into GBC-SD cell lines. Growing transfected cells were selected by G418. The presence and expression of exogenous p53 gene was detected by PCR, RT-PCR and Western blot. The cellular proliferating ability was assessed using the cell growth curve and cloning assay. The xenograft in nude mice was performed to examine the effect of tumorigenicity. Results: P53 protein overexpression was showed in GBC-SD cell lines. A transversion of TAC→AAC at codon 126 of exon 5 was confirmed. PCR, RT-PCR and Western blot showed exogenous p53 gene had successfully transfected into GBC-SD cells and obtained high expression. The growth and proliferation of the cells were greatly decreased, and the tumorigenicity was significantly inhibited after transfection wtp53. Conclusion: The expression of exogenous wild-type p53 gene could effectively inhibit the growth of gallbladder cancer GBC-SD cells in vitro and in vivo.