Yincheng Zhang1, Jun Liu, Fan Xue, Jiawei Li. 1. Department of Maxillofacial Surgery, Stomatological College of Xi'an Jiaotong University, Xi'an, China. xuefan99@yahoo.com
Abstract
OBJECTIVE: 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. METHODS: 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. RESULTS: 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. CONCLUSION: 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.
OBJECTIVE: This study aimed to determine the growth inhibitory effects of transfecting the wild-type p16 gene into humansalivary adenoid cystic carcinoma (SACC) cells in vitro. METHODS: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. RESULTS: 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. CONCLUSION: 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.
Authors: João Augusto Vianna Goulart-Filho; Victor Angelo Martins Montalli; Fabrício Passador-Santos; Ney Soares de Araújo; Vera Cavalcanti de Araújo Journal: Diagn Pathol Date: 2019-02-08 Impact factor: 2.644