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Rb/E2F4 and Smad2/3 link survivin to TGF-β-induced apoptosis and tumor progression

Oncogene volume 27, pages 5326–5338 (2008)Cite this article

Abstract

Survivin is a prosurvival protein overexpressed in many cancers through mechanisms that remain poorly explored, and is implicated in control of tumor progression and resistance to cancer chemotherapeutics. Here, we report a critical role for survivin in the induction of apoptosis by transforming growth factor-β (TGF-β). We show that TGF-β rapidly downregulates survivin expression in prostate epithelial cells, through a unique mechanism of transcriptional suppression involving Smads 2 and 3, Rb/E2F4, and the cell-cycle repressor elements CDE and CHR. This TGF-β response is triggered through a Smad2/3-dependent hypophosphorylation of Rb and the subsequent association of the Rb/E2F4 repressive complex to CDE/CHR elements in the proximal region of the survivin promoter. Viral-mediated gene delivery experiments, involving overexpressing or silencing survivin, reveal critical roles of survivin in apoptosis induced by TGF-β alone or in cooperation with cancer therapeutic agents. We propose a novel TGF-β/Rb/survivin axis with a putative role in the functional switch of TGF-β from tumor suppressor to tumor promoter.

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Abbreviations

3TP-Lux:

3TP-luciferase

CA-ALK5:

constitutively active ALK5

Cdk:

cyclin-dependent kinase

DC-FBS:

dextran charcoal-treated FBS

DMEM/F-12:

Dulbecco's modified Eagle's medium/Ham's F-12

DNAP:

DNA pull-down assay

EMSA:

electrophoretic mobility shift assay

FACS:

fluorescence-activated cell sorting

FBS:

fetal bovine serum

HEK293T:

human embryonic kidney cell line 293 inserted with SV40 T antigen

IAP:

inhibitor of apoptosis

P-Rb:

phosphorylated Rb

P-Smad:

phosphorylated Smad

Rb:

retinoblastoma protein

R-Smad:

receptor-regulated Smad

SAC:

a 32-bp survivin promoter region (−50/−19) containing SBE2, CDE and CHR elements

SBE:

Smad-binding element

shRNA:

short-hairpin RNA

TβRI:

TGF-β receptor type I

TβRII:

TGF-β receptor type II

TGF-β:

transforming growth factor-β

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Acknowledgements

This research was supported by the Gene Expression and Genotyping Facility, the Flow Cytometry Core Facility of the Case Comprehensive Cancer Center (P30 CA43703). Grant support: NCI grants R01CA092102 and R01CA102074 (to D Danielpour).

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Authors and Affiliations

  1. Division of General Medical Sciences—Oncology, Case School of Medicine, Case Western Reserve University, Cleveland, OH, USA

    J Yang, K Song, T L Krebs & D Danielpour

  2. Department of Biochemistry, Case School of Medicine, Case Western Reserve University, Cleveland, OH, USA

    J Yang

  3. Department of Pathology, Case School of Medicine, Case Western Reserve University, Cleveland, OH, USA

    M W Jackson

  4. Department of Pharmacology, Case School of Medicine, Case Western Reserve University, Cleveland, OH, USA

    D Danielpour

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  1. J Yang
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Correspondence to D Danielpour.

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Supplementary Information accompanies the paper on the Oncogene website (http://www.nature.com/onc)

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Yang, J., Song, K., Krebs, T. et al. Rb/E2F4 and Smad2/3 link survivin to TGF-β-induced apoptosis and tumor progression. Oncogene 27, 5326–5338 (2008). https://doi.org/10.1038/onc.2008.165

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