TY - JOUR
T1 - Activation of caspase-3 and cleavage of Rb are associated with p16-mediated apoptosis in human non-small cell lung cancer cells
AU - Katsuda, Koh
AU - Kataoka, Masafumi
AU - Uno, Futoshi
AU - Murakami, Takayoshi
AU - Kondo, Tadashi
AU - Roth, Jack A.
AU - Tanaka, Noriaki
AU - Fujiwara, Toshiyoshi
N1 - Funding Information:
We thank Dr Ta-Jen Liu for the gift of Adv/p16. We also thank Tohru Tanida, Yoshiko Takata and Kyoko Nasu for their technical support. This work was supported in part by grants from the Ministry of Education, Science, and Culture, Japan; by grants from the Ministry of Health and Welfare, Japan (Health Sciences Research Grants [Research on Human Genome and Gene Therapy]); by grants from the National Cancer Institute and the National Institutes of Health SPORE (2P50-CA70970-04) [JA Roth]; (P01 CA78778-01A1) [JA Roth]; by gifts to the Division of Surgery, from Tenneco and Exxon for the Core Laboratory Facility; by the UT M.D. Anderson Cancer Center Support Core Grant (CA 16672); by a grant from the Tobacco Settlement Funds as appropriate by the Texas State Legislature (Project 8), and The W. M. Keck Center for Cancer Gene Therapy [JA Roth].
PY - 2002
Y1 - 2002
N2 - The p16 tumor suppressor gene is frequently inactivated in human cancer tissues and cell lines. We previously reported that wild-type p16 expression from an adenovirus vector (Adv/p16) induced p53-dependent apoptotic cell death in non-small cell lung cancer (NSCLC) cell lines. Here we show the potential mechanism of apoptosis induced by Adv/p16 infection. Infection of human NSCLC cell line A549, which carries the wild-type p53 gene, with Adv/p16 resulted in activation of caspase-3, accompanied by the cleavage of its substrate poly (ADP-ribose) polymerase (PARP), on day 3 of infection. The retinoblastoma (Rb) cell cycle regulator protein was also cleaved after activation of caspase-3; when the levels of Rb significantly diminished, apoptosis began. When A549 cells were pretreated with the caspase-inhibitory peptide N-acetyl-asp-Glu-Val-Asp-CHO (aldehyde) (Ac-DEVD-CHO), Adv/p16-mediated apoptosis and Rb cleavage were greatly inhibited. Furthermore, MDM2, a negative regulator of p53 expression was upregulated 3 days after Adv/p16 infection, and MDM2 was subsequently cleaved by caspase-3; MDM2 cleavage was inhibited by Ac-DEVD-CHO treatment. These data implied that cleavage of Rb, in addition to activation of caspase-3, represented a mechanism by which Adv/p16 induced apoptotic cell death in human NSCLC cells. Our results support the clinical relevance of Adv/p16 as a treatment for p16-null human NSCLC that express wild-type p53.
AB - The p16 tumor suppressor gene is frequently inactivated in human cancer tissues and cell lines. We previously reported that wild-type p16 expression from an adenovirus vector (Adv/p16) induced p53-dependent apoptotic cell death in non-small cell lung cancer (NSCLC) cell lines. Here we show the potential mechanism of apoptosis induced by Adv/p16 infection. Infection of human NSCLC cell line A549, which carries the wild-type p53 gene, with Adv/p16 resulted in activation of caspase-3, accompanied by the cleavage of its substrate poly (ADP-ribose) polymerase (PARP), on day 3 of infection. The retinoblastoma (Rb) cell cycle regulator protein was also cleaved after activation of caspase-3; when the levels of Rb significantly diminished, apoptosis began. When A549 cells were pretreated with the caspase-inhibitory peptide N-acetyl-asp-Glu-Val-Asp-CHO (aldehyde) (Ac-DEVD-CHO), Adv/p16-mediated apoptosis and Rb cleavage were greatly inhibited. Furthermore, MDM2, a negative regulator of p53 expression was upregulated 3 days after Adv/p16 infection, and MDM2 was subsequently cleaved by caspase-3; MDM2 cleavage was inhibited by Ac-DEVD-CHO treatment. These data implied that cleavage of Rb, in addition to activation of caspase-3, represented a mechanism by which Adv/p16 induced apoptotic cell death in human NSCLC cells. Our results support the clinical relevance of Adv/p16 as a treatment for p16-null human NSCLC that express wild-type p53.
KW - Adenovirus vector
KW - Apoptosis
KW - Caspase-3
KW - P16
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UR - http://www.scopus.com/inward/citedby.url?scp=85047699958&partnerID=8YFLogxK
U2 - 10.1038/sj.onc.1205272
DO - 10.1038/sj.onc.1205272
M3 - Article
C2 - 11960384
AN - SCOPUS:85047699958
SN - 0950-9232
VL - 21
SP - 2108
EP - 2113
JO - Oncogene
JF - Oncogene
IS - 13
ER -