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The effects of exogenous p53 overexpression on HPV‐immortalized and carcinogen transformed oral keratinocytes
Author(s) -
Yoo George H.,
Washington James,
Oliver Jeffrey,
Piechocki Marie,
Kim Harold,
FosterNora Jessica,
Shibuya Terry Y.,
Wilson Deborah R.,
Ensley John F.
Publication year - 2001
Publication title -
cancer
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 3.052
H-Index - 304
eISSN - 1097-0142
pISSN - 0008-543X
DOI - 10.1002/cncr.10210
Subject(s) - cell culture , immortalised cell line , cancer research , apoptosis , transfection , cell cycle , cell growth , carcinogen , biology , keratinocyte , cell , microbiology and biotechnology , biochemistry , genetics
BACKGROUND Overexpression of p53 in head and neck carcinoma cells has demonstrated tumor growth suppression using in vitro and in vivo models. The effects of exogenous overexpression of wild‐type p53 on human papilloma virus (HPV)–immortalized and carcinogen transformed oral keratinocytes were determined. METHODS The p53 gene was overexpressed in IHGK (immortalized human gingival keratinocyte), IHGKN [4‐(methylnitrosamino)‐1‐(3‐pyridyl)‐1‐butanone, (NNK)]‐carcinogen transformed keratinocytes, and two head and neck squamous carcinoma (HNSCC) cell lines, HN30 and HN12. The transfection efficiency, growth suppression, and inhibition of the cell cycle along with the induction of apoptosis were measured. RESULTS Transfections with adenoviruses were more efficient for IHGK cells than for IHGKN, HN12, and HN30 cells. Inhibition of proliferation in all cell lines was proportional to the viral particle to cell (VPC) ratios. IHGK cells were more sensitive to p53 than IHGKN cells. HN12 cells were more suppressed than HN30 cells. HN12 were the most suppressed at 72 hours whereas HN30 cells were most suppressed at 24 hours. Expression of exogenous p53‐induced G1 cell cycle arrest and p21 expression as VPC ratios increased in IHGK and IHGKN cell lines. Apoptosis also was induced in these cells by p53 as VPC increased. IHGK cells were more sensitive to p53‐induced growth inhibition, cell cycle regulation, p21 expression and apoptosis than IHGKN cells. HN12 (mutated p53 ) cells were more sensitive to p53 overexpression than HN30 (wild‐type p53 ) cells. Gene transfer and expression of exogenous p53 by using Ad‐p53 demonstrates suppressive effects on HPV immortalized and carcinogen transformed oral keratinocytes. CONCLUSIONS Cell cycle regulation by gene transfer is feasible in immortalized oral keratinocytes. Carcinogen transformed cells are less susceptible to the effects of p53 overexpression. Expression of exogenous p53 through p53 gene transfer can suppress HPV immortalization and carcinogen transformation in oral keratinocytes. The sensitivity of HNSCC cell lines to p53‐induced cell cycle regulation and apoptosis is variable and dependent on the cell line and duration of exposure. In vitro results using p53 gene transfer must be validated in clinical studies with patients at risk for HNSCC. Cancer 2002;94:159–66. © 2002 American Cancer Society.