Transfer of microRNA-25 by colorectal cancer cell-derived extracellular vesicles facilitates colorectal cancer development and metastasis

Shanchao Wang,Zeyan Zhang,Qianfu Gao

doi:10.1016/j.omtn.2020.11.018

Abstract

Cancer cell-derived extracellular vesicles (EVs) have been reported to promote the progression of colorectal cancer (CRC), although the regulatory mechanism remains uncharacterized. In this study, we investigated the role of microRNA-25 (miR-25)/sirtuin 6 (SIRT6) in the contribution of EVs derived from CRC cells to progression of CRC. In a co-culture system with EVs from HCT116 and NCM460 cells, the viability, migratory, and invasive properties of SW480 and SW620 cells were evaluated by cell counting kit-8 (CCK-8) and Transwell assays. Luciferase, chromatin immunoprecipitation (ChIP), and RNA immunoprecipitation (RIP) assays were conducted to verify the interaction among miR-25, SIRT6, lin-28 homologB (Lin28b), and neuropilin-1 (NRP-1). It was established that HCT116 cell-derived EVs promoted the malignant properties of SW480 cells and SW620 cells by delivering miR-25. SIRT6 was targeted by miR-25, whereas SIRT6 inhibited NRP-1 through downregulation of Lin28b. The tumor-bearing nude mouse experiments substantiated that HCT116 cell-derived EVs transferred miR-25 to facilitate tumor formation and metastasis by inhibiting SIRT6. In summary, our study clarifies the involvement of miR-25-targeted SIRT6 inhibition and SIRT6-mediated inhibition of the Lin28b/NRP-1 axis in CRC cell-derived EVs to CRC progression and metastasis.

Highlights

Colorectal cancer (CRC) is one of the leading causes of cancerrelated death globally, and the mortality of patients with CRC ranks fifth in China according to cancer statistics in 2015.1,2 improved screening and surveillance make great contributions to the early detection of CRC, minimal progress has been achieved in mitigating the devastating impact of CRC metastasis to the liver.[3]
Enrichment of miR-25 in CRC cell-derived Extracellular vesicles (EVs) and its transfer by EVs In the first phase of the study, clinical CRC tissues were collected and analyzed by quantitative reverse-transcriptase polymerase chain reaction, which showed higher miR-25 expression in cancer tissues than in the matched non-cancerous tissues, and that miR-25 expression was higher in cancer tissues of patients with metastasis than in those from patients without metastasis (Figure 1A)
To verify whether the miR-25 in EVs derived from CRC cells could affect the metastatic characteristics of cancer cells, we extracted EVs from NCM460 and HCT116 cells

Summary

Introduction

Colorectal cancer (CRC) is one of the leading causes of cancerrelated death globally, and the mortality of patients with CRC ranks fifth in China according to cancer statistics in 2015.1,2 improved screening and surveillance make great contributions to the early detection of CRC, minimal progress has been achieved in mitigating the devastating impact of CRC metastasis to the liver.[3] lung or liver metastases of CRC remain a major obstacle in clinical therapeutics.[4] more effective therapies for CRC are urgently needed. MiRNAs are small and non-coding regulatory RNAs, which have emerged as valuable biomarkers and therapeutic targets for CRC.[8] For example, miR-25 is highly expressed in CRC and is a marker of poor prognosis of patients with CRC.[9] Intriguingly, a major type of EVs, namely the exosomes, can transfer miR-25 from CRC cells to endothelial cells to promote the occurrence and metastasis of CRC.[10] the mechanism underlying this process remains largely unexplored

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Results

Conclusion