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Adenovirus‐mediated p16 gene transfer prevents drug‐induced cell death through G 1 arrest in human glioma cells
Author(s) -
Hama Seiji,
Heike Yuji,
Naruse Ichiro,
Takahashi Minako,
Yoshioka Hiroyuki,
Arita Kazunori,
Kurisu Kaoru,
Goldman Corey K.,
Curiel David T.,
Saijo Nagahiro
Publication year - 1998
Publication title -
international journal of cancer
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.475
H-Index - 234
eISSN - 1097-0215
pISSN - 0020-7136
DOI - 10.1002/(sici)1097-0215(19980703)77:1<47::aid-ijc9>3.0.co;2-#
Subject(s) - programmed cell death , trypan blue , cell cycle , cancer research , glioma , cell growth , microbiology and biotechnology , cell culture , cell , cell cycle checkpoint , biology , apoptosis , biochemistry , genetics
This study examined the effects of full‐length p16 gene transfer by recombinant adenovirus on cell growth and on sensitivity to CDDP or ACNU chemotherapies. We developed a recombinant adenovirus expressing the full‐length human p16 gene (AxCA‐ hp16 ) by the COS‐TPC method. AxCA‐ hp16 was infected into the p16 ‐null human glioma cell line, U251MG. AxCA‐ hp16 infection inhibited proliferation of U251MG cells. A proliferation assay employing MTT showed that AxCA‐ hp16 infection induced chemoresistance, preventing CDDP‐induced cell death (11‐ to 15‐fold) and ACNU‐induced cell death (80‐ to 92‐fold). In the absence of AxCA‐ hp16, cell death was induced with CDDP or ACNU at 3 to 5 days after treatment, as demonstrated by Trypan‐blue exclusion. Flow‐cytometric analysis showed that CDDP or ACNU arrested cells in the G 2 phase on day 1 and that cells re‐entered the cycle on day 3. However, the cells infected with AxCA‐ hp16 after CDDP or ACNU treatment showed G 1 arrest on day 5 after re‐entering the cycle from G 2 arrest on day 3. The cells infected with AxCA‐ hp16 before CDDP or ACNU treatment showed G 1 arrest over the 5 days after the infection. This study demonstrated that G 1 arrest induced with p16 ‐gene expression prevents ACNU‐ or CDDP‐induced cell death. The cell death induced by ACNU and CDDP therefore appears to occur in the phase after the G 1 /S check point. Int. J. Cancer 77:47–54, 1998.© 1998 Wiley‐Liss, Inc.