Adenovirus-mediated p16INK4a gene expression radiosensitizes non-small cell lung cancer cells in a p53-dependent manner

Kawabe, Shinichiro; Roth, Jack A.; Wilson, Debra Rose; Meyn, Raymond E.

doi:10.1038/sj.onc.1203935

Cited by 42 publications

(29 citation statements)

References 21 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…We found that Adv/p16 infection induced cleavage of MDM2 and upregulation of p53 prior to triggering apoptosis. Others have shown that accumulation of the p53 protein by Adv/p16 sensitizes NSCLC to irradiation (Kawabe et al, 2000). These observations support the possibility that caspase-3 activation-induced cleavage of MDM2 results in transient p53 activation, leading to induction of apoptosis in Adv/p16-infected A549 cells; direct interactions among these molecules, however, should be further examined.…”

supporting

confidence: 45%

Activation of caspase-3 and cleavage of Rb are associated with p16-mediated apoptosis in human non-small cell lung cancer cells

et al. 2002

Self Cite

View full text Add to dashboard Cite

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 signi®cantly 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.

show abstract

supporting

confidence: 45%

Activation of caspase-3 and cleavage of Rb are associated with p16-mediated apoptosis in human non-small cell lung cancer cells

et al. 2002

Self Cite

View full text Add to dashboard Cite

show abstract

“…The results indicate that the cell survival of these cancer cell lines was substantially decreased by Ax-hp16 infection (data not shown) (Kawabe et al, 2000;Yokozaki, 2000). These findings suggest that adenovirus-mediated p16 gene transfer increased radiosensitivity via nonapoptotic cell killing in these other human cancer cell lines.…”

Section: Discussionmentioning

confidence: 82%

“…Previous findings indicate that p16-dependent downmodulation of pRb levels in the presence of overexpressed wild-type p53 causes apoptosis only in tumour cells, whereas normal cells arrest in G1 (Sandig et al, 1997). Kawabe et al (2000) U251MG cells Figure 4 Effect of p16 or p21 transduction with or without irradiation on nuclear morphologic changes. U251MG or D54MG cells were treated as described in the TUNEL assay, and nuclear morphologic changes in these treated cells were evaluated using Giemsa staining 7 days after irradiation.…”

Section: Discussionmentioning

confidence: 99%

“…However, Ax-hp16-infected U251MG and D54MG cells were arrested in the G1 phase during the 6 days following irradiation, suggesting that the induction of G1 arrest following irradiation increases the radiosensitivity of these cells. Kawabe et al (2000) reported that p16 gene transfer enhanced apoptotic cell death for cells that have wt-p53, but not for p53-mutated or p53-deleted cells. The present findings indicate that p16 enhanced radiosensitivity via increased cell death, and that this enhancement did not depend on p53 status.…”

Section: Discussionmentioning

confidence: 99%

“…Adenovirus-mediated gene transfer of wild-type p16 following irradiation has previously been shown (by clonogenic survival assay) to enhance radiosensitivity in lung non-small-cell cancer cells (Kawabe et al, 2000). The results of clonogenic survival assays clearly indicate that cells synchronised at the late S and early G1 phase are the most radioresistant, whereas cells synchronized at the M phase or at the late G1 to early S phase are the most radiosensitive (Terasima and Tolmach, 1963;Okada, 1970).…”

mentioning

confidence: 95%

See 2 more Smart Citations

p16 gene transfer increases cell killing with abnormal nucleation after ionising radiation in glioma cells

et al. 2003

View full text Add to dashboard Cite

It is well established that cells synchronised at the G1 -S phase are highly radiosensitive. In this study, p16-null human glioma cell lines were induced into G1 cell cycle arrest by adenovirus-mediated p16 gene transfer, and examined for radiation-induced cell killing. Clonogenic analysis and trypan blue extraction test showed that the p16 gene transfer enhanced radiation-induced cell killing in p16-null glioma cell lines. TUNEL assays and pulse-field gel electrophoresis confirmed that the radiation-induced cell killing of p16-transfected cells could be caused by a nonapoptotic mechanism. Gimsa staining demonstrated that irradiation alone or Ax-mock infection plus irradiation results in a slight increase in the frequency of cells with abnormal nucleus, compared to unirradiated uninfected or Ax-mock infected cells. However, Ax-hp16 or Ax-hp21 infection alone modestly increased the frequency of cells with abnormal nucleus (especially bi-and multinucleation), and 4-Gy irradiation of Ax-hp16 or Ax-hp21 infected cells substantially enhanced this frequency. These results suggest that there exists some unknown interaction between radiation and p16 in cytoplasm/ membranes, which decreases cytokinesis and promotes abnormal nucleation. Thus, p16 expression prevented radiation-induced apoptosis by promoting abnormal nucleation, thereby leading to another mode of cell death.

show abstract

p53, p16 and cyclin D1: Molecular determinants of radiotherapy treatment response in oral carcinoma

Jayasurya

Francis

Kannan

et al. 2004

Intl Journal of Cancer

View full text Add to dashboard Cite

Management of oral cancer by radiotherapy has witnessed promising advances in the past few years, with patient-tailored radio fractionation regimens. Different fractionation schedules, conventional and altered regimes, have been used in curative radiotherapy. Although contribution of biological markers on radio response has been evaluated, its unique influence on various radio fractionation schemes has not been accounted so far. Our study analyses a set of proteins that previously demonstrated radio response influence for their possible prognostic value in decision-making process between the respective fractionation schemes. Expression patterns of regulatory proteins such as p53, cyclin D1, p16, Cdk4, p21, Rb, bcl-2 and PCNA were determined by immunohistochemistry utilizing monoclonal antibodies in 125 patients who received curative radiotherapy dose. Among these 125 patients, 90 (72%) received altered fractionation, whereas 35 (28%) received conventional fractionation. p53 over-expression correlated with local treatment failure among the patients treated with conventional fractionation whereas cyclin D1 over-expression and p16 underexpression were associated with local treatment failure as well as overall survival in altered fractionation treated cases. Our findings suggest that wild-type p53 status may be an important parameter for achieving high local control in those patients undergoing conventional fractionation, where as intact p16 and cyclin D1 status may be beneficial for effective local control in patients who are treated with altered fractionation. Furthermore, it can be assumed that conventional fractionation employs p53-mediated apoptosis, whereas altered fractionation activates the functional G1 cell-cycle checkpoint for tumor growth suppression.

show abstract

Adenovirus-mediated p16INK4a gene expression radiosensitizes non-small cell lung cancer cells in a p53-dependent manner

Cited by 42 publications

References 21 publications

Activation of caspase-3 and cleavage of Rb are associated with p16-mediated apoptosis in human non-small cell lung cancer cells

Activation of caspase-3 and cleavage of Rb are associated with p16-mediated apoptosis in human non-small cell lung cancer cells

p16 gene transfer increases cell killing with abnormal nucleation after ionising radiation in glioma cells

p53, p16 and cyclin D1: Molecular determinants of radiotherapy treatment response in oral carcinoma

Contact Info

Product

Resources

About