The role of E26 transformation-specific variant transcription factor 5 in colorectal cancer cell proliferation and cell cycle progression

Yi Peng,Yaqi Zhang,Ren Zhao,Kun Liu,Xi Cheng,Zijia Song,Haoran Feng,Changgang Wang

doi:10.1038/s41419-021-03717-5

Yi Peng, Yaqi Zhang + Show 6 more

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https://doi.org/10.1038/s41419-021-03717-5

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Abstract

E26 transformation-specific variant transcription factor 5 (ETV5) contributes to tumor growth and progression and promotes colorectal cancer (CRC) angiogenesis. Previous studies indicate that ETV5 may regulate the cell cycle, but its detailed mechanism remain unclear. Gene Ontology (GO) analysis of RNA-seq data revealed that ETV5 possibly regulates the cell cycle in CRC. Here, in vitro and in vivo experiments were performed to verify that ETV5 promoted tumor progression and influenced cell cycle G1/S transition. Cell cycle PCR array and co-immunoprecipitation (Co-IP) helped identify the p21-CDKs pathway. Chromatin immunoprecipitation (ChIP) and luciferase reporter assays were performed to determine whether ETV5 binds to the p21 promoter. ETV5 and p21 were detected by immunohistochemistry, and the effects of their expression on CRC patients were evaluated. ETV5 upregulation enhanced tumor proliferative capacity and promoted G1 phase transfer to the S phase. ETV5 knockdown slowed the growth of CRC cells and repressed the G1/S transition. We also found p21 as a downstream target of ETV5. p21 knockdown resulted in faster CRC cell growth and in more cells being driven from the G0/1 phase into the S phase. Co-IP experiments showed that p21 banding to CDK2, CDK4, and CDK6 inhibited p130 phosphorylation. Using the ChIP and luciferase reporter assay, we confirmed that ETV5 bound to the p21 promoter and repressed p21 expression. CRC patients with high ETV5 expression and low p21 expression showed the worst prognosis. Finally, by targeting p21 to regulate CDK function, ETV5 also changed drug-sensitivity to palbociclib and dinaciclib. In conclusion, ETV5 promoted cell cycle G1/S transition through transcriptional inhibition of p21, thereby accelerating tumor growth. Moreover, ETV5 changed drug-sensitivity to palbociclib and dinaciclib. Therefore, therapeutic regimens targeting ETV5 may be promising in improving the efficacy of target-CDK treatment in CRC.

Highlights

Colorectal cancer (CRC) represents 10.2% of all new cancer cases, and is the third most prevalent cancer worldwide
Based on the Gene Ontology (GO) analysis of RNA-seq data from our last study[19], we found that E26 transformation-specific variant transcription factor 5 (ETV5) was related to cell cycle regulation via many pathways (Supplement Fig. 1)
Co-IP was conducted, and the results showed that p21 could combine CDK2, CDK4, and CDK6. p21 increased when ETV5 was knocked down and bound more cyclindependent kinase (CDK), and reverse outcomes were observed in ETV5-overexpressing cells (Fig. 4C, D)

Summary

Introduction

Colorectal cancer (CRC) represents 10.2% of all new cancer cases, and is the third most prevalent cancer worldwide. The ETS family regulates the expression of genes involved in normal cell development, proliferation, differentiation, angiogenesis, and apoptosis. Dysregulation of these transcription factors facilitates cell proliferation in cancers, and several members participate in invasion and metastasis by activating certain gene transcriptions[8]. ERG aberrant expression occurs in 50% of prostate tumors, and that of ETS family transcription factors ETV1 and ETV4 occurs in another 10% of cases. These three ETS factors are thought to promote tumorigenesis in the majority of prostate cancers[9]. ERG and ETV6 promote thrombocytopenia and leukemia and are crucial for its maintenance[11,12]

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