p16 tumor suppressor therapy in salivary adenoid cystic carcinoma cell line SACC83.
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.


CONCLUSIONS
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.
Zhang
Yincheng
Y
Department of Maxillofacial Surgery, Stomatological College of Xi'an Jiaotong University, Xi'an, China. xuefan99@yahoo.com
Liu
Jun
J
Xue
Fan
F
Li
Jiawei
J
D003528:Carcinoma, Adenoid Cystic / Q000235:genetics / Q000628:therapy*; D002453:Cell Cycle; D045744:Cell Line, Tumor; D049109:Cell Proliferation; D004396:Coloring Agents / Q000378:metabolism; D004247:DNA / Q000378:metabolism; D005434:Flow Cytometry; D019942:Genes, p16 / Q000502:physiology*; D015316:Genetic Therapy; D006801:Humans; D016014:Linear Models; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012468:Salivary Gland Neoplasms / Q000235:genetics / Q000628:therapy*; D013778:Tetrazolium Salts / Q000378:metabolism; D013844:Thiazoles / Q000378:metabolism; D014162:Transfection
D004396:Coloring Agents; D013778:Tetrazolium Salts; D013844:Thiazoles; D004247:DNA; C022616:thiazolyl blue