Adenovirus-mediated p16 gene transfer prevents drug-induced cell death through G1 arrest in human glioma cells.
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 G2 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 G1 arrest on day 5 after re-entering the cycle from G2 arrest on day 3. The cells infected with AxCA-hp16 before CDDP or ACNU treatment showed G1 arrest over the 5 days after the infection. This study demonstrated that G1 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 G1/S check point.
Hama
S
S
Pharmacology Division, National Cancer Center Research Institute, Tokyo, Japan.
Heike
Y
Y
Naruse
I
I
Takahashi
M
M
Yoshioka
H
H
Arita
K
K
Kurisu
K
K
Goldman
C K
CK
Curiel
D T
DT
Saijo
N
N
D000256:Adenoviridae; D000970:Antineoplastic Agents / Q000494:pharmacology*; D016923:Cell Death / Q000187:drug effects / Q000235:genetics; D019941:Cyclin-Dependent Kinase Inhibitor p16 / Q000235:genetics*; D019008:Drug Resistance, Neoplasm / Q000235:genetics*; D016193:G1 Phase / Q000235:genetics; D018014:Gene Transfer Techniques; D005822:Genetic Vectors; D005910:Glioma / Q000235:genetics* / Q000473:pathology*; D006801:Humans; D015376:Nimustine / Q000494:pharmacology*; D014407:Tumor Cells, Cultured
D000970:Antineoplastic Agents; D019941:Cyclin-Dependent Kinase Inhibitor p16; D015376:Nimustine