Adenovirus-mediated p53 gene therapy in pediatric soft-tissue sarcoma cell lines: sensitization to cisplatin and doxorubicin

Ganjavi, Hooman; Gee, Matthew F.W.; Narendran, Aru; Freedman, Melvin H.; Malkin, David

doi:10.1038/sj.cgt.7700798

Cited by 35 publications

(23 citation statements)

References 70 publications

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“…Other possible determinants of the sensitivity to 2-DG are p53 and HIF-1, which regulate the glycolytic phenotype. p53 is not functional in any of the 3 sensitive alveolar rhabdomyosarcoma cell lines tested (29,30), which would agree with data that indicate that p53 protects cells from metabolic stress. However, RD cells are also deficient in p53 but still insensitive.…”

Section: Discussionsupporting

confidence: 80%

“…Apoptosis induced by glucose deprivation in hematopoietic cells is inhibited by overexpression of Bcl-2 or Bcl-x L , and it has been shown to be mediated by Noxa, Puma, or Bim (20,25). We could not detect induction of Puma, possibly because this protein is usually induced in a p53-dependent manner, but the alveolar rhabdomyosarcoma cells used in this study are deficient in p53 (29,30). Consistent with data that suggest that death induced by 2-DG is due to ER stress, we observed induction of Bim, which mediates death induced by ER stress in some systems (37).…”

Section: Discussioncontrasting

confidence: 46%

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2-Deoxyglucose Induces Noxa-Dependent Apoptosis in Alveolar Rhabdomyosarcoma

et al. 2011

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Alveolar and embryonal rhabdomyosarcomas are childhood tumors that do not respond well to current chemotherapies. Here, we report that the glycolytic inhibitor 2-deoxyglucose (2-DG) can efficiently promote cell death in alveolar, but not embryonal, rhabdomyosarcoma cell lines. Notably, 2-DG also induced cell differentiation accompanied by downregulation of PAX3/FOXO1a, the chromosome translocation-encoded fusion protein that is a central oncogenic driver in this disease. Cell death triggered by 2-DG was associated with its ability to activate Bax and Bak. Overexpression of the antiapoptotic Bcl-2 homologues Bcl-x L and Mcl-1 prevented apoptosis, indicating that cell death proceeds through the mitochondrial pathway. Mechanistic investigations indicated that Mcl-1 downregulation and Noxa upregulation were critical for 2-DG-induced apoptosis. In addition, 2-DG promoted eIF2a phosphorylation and inactivation of the mTOR pathway. Mcl-1 loss and cell death were prevented by downregulation of the endoplasmic reticulum (ER) stress-induced protein ATF4 and by incubating cells in the presence of mannose, which reverted 2-DG-induced ER stress but not ATP depletion. Thus, energetic stresses created by 2-DG were not the primary cause of cell death. Together, our findings suggest that glycolysis inhibitors such as 2-DG may be highly effective in treating alveolar rhabdomyosarcoma and that Noxa could offer a prognostic marker to monitor the efficacy of such agents. Cancer Res; 71(21); 6796-806. Ó2011 AACR.

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Section: Discussionsupporting

confidence: 80%

Section: Discussioncontrasting

confidence: 46%

2-Deoxyglucose Induces Noxa-Dependent Apoptosis in Alveolar Rhabdomyosarcoma

et al. 2011

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“…We observed that all cell lines tested were resistant to hypoxia-induced apoptosis (Figure 1a). For further experiments we selected a p53 wild type (A204 RMS) and a p53-deficient (A673 ES) cell line (Ganjavi et al, 2005). Additionally, analysis by flow cytometry using Annexin V-fluorescein isothiocyanate (FITC)/ propidium-iodide (PI) double staining revealed that hypoxia also did not induce necrotic cell death in both A204 RMS and A673 ES cells (Figure 1b).…”

Section: Resistance Of A204 Rms and A673 Es Cells To Hypoxiainduced Amentioning

confidence: 99%

Role of hypoxia inducible factor-1 alpha in modulation of apoptosis resistance

et al. 2006

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Hypoxia inducible factor-1 (HIF-1) is the major transcription factor and key regulator of adoptive responses to hypoxia. Although it usually promotes tumor cell survival under hypoxia, it has also been implied to trigger apoptosis. Although the impact of hypoxia has been extensively studied in many adult solid tumors, its role in most childhood tumors, for example, in rhabdomyosarcoma (RMS) or Ewing sarcoma (ES), has not yet been addressed. Here, we report that hypoxia protects A204 RMS and A673 ES cells against anticancer drug-or tumor necrosis factor-related apoptosis-inducing ligandinduced apoptosis and that Hif-1a plays a key role in conferring apoptosis resistance under hypoxia. Although a functional HIF-1 pathway and proapoptotic proteins such as p53 and Bcl-2/E1B 19 kDa interacting protein 3 were activated under hypoxia in both A204 RMS and A673 ES cells, these cells remained refractory to apoptosis. Concomitant analysis of antiapoptotic proteins revealed that hypoxia induced expression of Bcl-2 and inhibitor of apoptosis proteins (IAP)-2 as well as proteins associated with anaerobic metabolism such as the glucose transporter protein GLUT-1 and the glycolytic enzyme Aldolase A. Specific downregulation of Hif-1a by RNA interference significantly enhanced apoptosis under hypoxia by preventing the hypoxia-mediated increase in GLUT-1 expression without altering expression levels of the antiapoptotic proteins Bcl-2 or cIAP-2. Moreover, glucose deprivation-induced apoptosis of A204 RMS and A673 ES cells was inhibited under hypoxic conditions in a Hif-1a-dependent manner. As GLUT-1 was induced via Hif-1a under hypoxia in A204 RMS and A673 ES, these findings suggest that the Hif-1a-mediated increase in glucose uptake plays an important role in conferring apoptosis resistance. Thus, hypoxia-inducible genes may represent novel targets for therapeutic intervention in some pediatric tumors, which warrants further investigation.

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“…Mutations in the p53 gene are among the most frequent somatic genetic changes observed in human cancers, and consequently, p53 mutational status has been extensively studied for its correlation with tumor grade, patient survival, and treatment outcome in many cancers (10). The results of these studies have, however, been inconsistent with some showing a positive correlation of p53 mutations with tumor growth, progression, and drug resistance (11)(12)(13)(14), whereas others showed the mutant p53 status to be deleterious to survival and to favor drug sensitivity (15)(16)(17). The findings indicate a high level of complexity in the role of p53 in human cancer and suggest that multiple upstream and downstream pathways of p53 may interact to mediate its effects in this disease.…”

Section: Introductionmentioning

confidence: 76%

Identification and Functional Characterization of the Human Glutathione S-Transferase P1 Gene as a Novel Transcriptional Target of the p53 Tumor Suppressor Gene

Stephenson

Cao

et al. 2008

Molecular Cancer Research

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The glutathione S-transferase P1 (GSTP1) is involved in multiple cellular functions, including phase II metabolism, stress response, signaling, and apoptosis. The mechanisms underlying the significantly high GSTP1 expression in many human tumors are, however, currently not well understood. We report here that the GSTP1 gene is a heretofore unrecognized downstream transcriptional target of the tumor suppressor p53. We identified a p53-binding motif comprising two consecutive half-sites located in intron 4 of the GSTP1 gene and is highly homologous to consensus p53-binding motifs in other p53-responsive genes. Using a combination of electrophoretic mobility shift assay and DNase I footprinting analyses, we showed that wild-type p53 protein binds to the GSTP1 p53 motif and luciferase reporter assays showed the motif to be transcriptionally functional in human tumor cells. In a temperature-sensitive p53-mutant cells, levels of both p21/WAF1 and GSTP1 gene transcripts increased time dependently when cells were switched from the inactive mutant state to the wild-type p53 state. Small interfering RNA -mediated reduction of p53 expression resulted in a specific decrease in GSTP1 expression and in tumor cells with mutated p53; adenovirally mediated expression of wild-type p53 increased GSTP1 expression significantly. In a panel of early-passage brain tumor cultures from patients, high levels of GSTP1 transcripts and protein were associated with wild-type p53 and, conversely, low GSTP1 levels with mutant p53. p53 expression knockdown by small interfering RNA increased cisplatin sensitivity. The ability of wild-type p53 to transcriptionally activate the human GSTP1 gene defines a novel mechanism of protecting the genome and, potentially, of tumor drug resistance. (Mol Cancer Res 2008;6(5):843 -50)

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Adenovirus-mediated p53 gene therapy in pediatric soft-tissue sarcoma cell lines: sensitization to cisplatin and doxorubicin

Cited by 35 publications

References 70 publications

2-Deoxyglucose Induces Noxa-Dependent Apoptosis in Alveolar Rhabdomyosarcoma

2-Deoxyglucose Induces Noxa-Dependent Apoptosis in Alveolar Rhabdomyosarcoma

Role of hypoxia inducible factor-1 alpha in modulation of apoptosis resistance

Identification and Functional Characterization of the Human Glutathione S-Transferase P1 Gene as a Novel Transcriptional Target of the p53 Tumor Suppressor Gene

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