Myc down-regulation affects cyclin D1/cdk4 activity and induces apoptosis via Smac/Diablo pathway in an astrocytoma cell line

D. Amendola, M. De Salvo, R. Marchese, C. Verga Falzacappa, A. Stigliano, E. Carico, E. Brunetti, M. Moscarini, B. Bucci

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Abstract

Objectives: We investigated the antiproliferative effect of Myc down-regulation via cell proliferation inhibition, cell cycle perturbation and apoptosis in two human astrocytoma models (T98G and ADF) steadily expressing an inducible c-myc Anti-sense RNA. Materials and methods: Cell growth experiments were performed using the trypan blue dye exclusion test and cell cycle analysis was evaluated by flow cytometry. Cell cycle molecules were detected by Western blot analysis, co-immunoprecipitation and reverse transcription-polymerase chain reaction assays. Results: We showed that Myc down-regulation in astrocytoma cells led to G1 accumulation and an inhibition of cell proliferation characterized by S phase delay. Co-immunoprecipitation experiments detected formation of inactive cyclin D1/cdk4 complexes as evaluated by presence of an active unphosphorylated form of retinoblastoma protein, the best characterized target substrate for cyclin D1/cdk4 complex, in ADF pINDc-myc anti-sense 7 cells. We also found that either p57Kip2 "apice" or p27Kip1 "apice" inhibitors bound to cyclin D1/cdk4 complex, thus, suggesting that they cooperated to inhibit the activity of cyclin D1/cdk4. Moreover, c-Myc down-regulation led to activation of the apoptotic mitochondrial pathway, characterized by release of cytochrome c and Smac/Diablo proteins and by reduction of c-IAP levels through activation of proteasome-mediated protein degradation system. Conclusions: Our results suggest that c-Myc could be considered as a good target for the study of new approaches in anticancer astrocytoma treatment.

Original languageEnglish
Pages (from-to)94-109
Number of pages16
JournalCell Proliferation
Volume42
Issue number1
DOIs
Publication statusPublished - Feb 2009

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Cyclin D1
Astrocytoma
Down-Regulation
Apoptosis
Cell Line
Cell Cycle
Immunoprecipitation
Cell Proliferation
Antisense RNA
Retinoblastoma Protein
Trypan Blue
Proteasome Endopeptidase Complex
Cytochromes c
S Phase
Proteolysis
Reverse Transcription
Flow Cytometry
Coloring Agents
Western Blotting
Polymerase Chain Reaction

ASJC Scopus subject areas

  • Cell Biology

Cite this

Amendola, D., De Salvo, M., Marchese, R., Falzacappa, C. V., Stigliano, A., Carico, E., ... Bucci, B. (2009). Myc down-regulation affects cyclin D1/cdk4 activity and induces apoptosis via Smac/Diablo pathway in an astrocytoma cell line. Cell Proliferation, 42(1), 94-109. https://doi.org/10.1111/j.1365-2184.2008.00576.x

Myc down-regulation affects cyclin D1/cdk4 activity and induces apoptosis via Smac/Diablo pathway in an astrocytoma cell line. / Amendola, D.; De Salvo, M.; Marchese, R.; Falzacappa, C. Verga; Stigliano, A.; Carico, E.; Brunetti, E.; Moscarini, M.; Bucci, B.

In: Cell Proliferation, Vol. 42, No. 1, 02.2009, p. 94-109.

Research output: Contribution to journalArticle

Amendola, D, De Salvo, M, Marchese, R, Falzacappa, CV, Stigliano, A, Carico, E, Brunetti, E, Moscarini, M & Bucci, B 2009, 'Myc down-regulation affects cyclin D1/cdk4 activity and induces apoptosis via Smac/Diablo pathway in an astrocytoma cell line', Cell Proliferation, vol. 42, no. 1, pp. 94-109. https://doi.org/10.1111/j.1365-2184.2008.00576.x
Amendola, D. ; De Salvo, M. ; Marchese, R. ; Falzacappa, C. Verga ; Stigliano, A. ; Carico, E. ; Brunetti, E. ; Moscarini, M. ; Bucci, B. / Myc down-regulation affects cyclin D1/cdk4 activity and induces apoptosis via Smac/Diablo pathway in an astrocytoma cell line. In: Cell Proliferation. 2009 ; Vol. 42, No. 1. pp. 94-109.
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AU - Stigliano, A.

AU - Carico, E.

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