P16 tumor suppressor therapy in salivary adenoid cystic carcinoma cell line SACC83

Abstract

This study aimed to determine the growth inhibitory effects of transfecting the wild-type p16 gene into human salivary adenoid cystic carcinoma (SACC) cells in vitro. p16 cDNA was cloned into pDOR-neo plasmid by recombination technique, and the human eukaryotic expression vector, pDOR-p16 , was constructed. P16 gene was transfected into SACC83 cells using the liposome method, and the expression of the transfected genes was detected by RT-PCR. The efficacy of transfection was determined by MTT-based colorimetric formats. The DNA metabolic rate of transfected cells was measured by 3 H-TdR incorporation. To describe the cell division phase, the cell cycles of the p16 -transfected gene cells and untransfected cells were analyzed by flow cytometry. The growths in vitro of the p16-transfected cells were inhibited significantly. 3H-TdR incorporation showed a decreased DNA metabolic rate of p16-transfected cells, and flow cytometry suggested a significant increase of the cells in the G1 phase and decrease in the S phase. The study confirmed the reversal effect of wild-type p16 on malignant phenotype of the salivary adenoid cystic carcinoma and provides valuable data for further clinical trial of gene therapy with p16.