Exosomal linc-ROR mediates crosstalk between cancer cells and adipocytes to promote tumor growth in pancreatic cancer

Zhaowei Sun; Dong Sun; Yujie Feng; Bingyuan Zhang; Peng Sun; Bin Zhou; Lutao Du; Yunshan Wang; Zhiyao Fan; Jian Yang; Yongzheng Li; Sanyuan Hu; Hanxiang Zhan

doi:10.1016/j.omtn.2021.06.001

Exosomal linc-ROR mediates crosstalk between cancer cells and adipocytes to promote tumor growth in pancreatic cancer

Mol Ther Nucleic Acids. 2021 Jun 4:26:253-268. doi: 10.1016/j.omtn.2021.06.001. eCollection 2021 Dec 3.

Authors

Zhaowei Sun^{1

2}, Dong Sun¹, Yujie Feng², Bingyuan Zhang², Peng Sun³, Bin Zhou², Lutao Du^{4

5

6}, Yunshan Wang^{4

5

6}, Zhiyao Fan¹, Jian Yang¹, Yongzheng Li¹, Sanyuan Hu⁷, Hanxiang Zhan¹

Affiliations

¹ Division of Pancreatic Surgery, Department of General Surgery, Qilu Hospital, Cheeloo College of Medicine, Shandong University, Jinan, Shandong, China.
² Department of Hepatobiliary and Pancreatic Surgery, The Affiliated Hospital of Qingdao University, Qingdao, China.
³ Department of Liver Transplant, The Affiliated Hospital of Qingdao University, Qingdao, China.
⁴ Department of Clinical Laboratory, The Second Hospital, Cheeloo College of Medicine, Shandong University, Jinan, China.
⁵ Shandong Engineering & Technology Research Center for Tumor Marker Detection, Jinan, China.
⁶ Shandong Provincial Clinical Medicine Research Center for Clinical Laboratory, Jinan, China.
⁷ Department of General Surgery, The First Affiliated Hospital of Shandong First Medical University, Jinan, China.

Abstract

Exosomes are emerging as important mediators of the crosstalk between tumor cells and stromal cells in the microenvironment. However, the underlying molecular mechanism of pancreatic cancer (PC)-derived exosomes in the progression of the tumor microenvironment (TME) and crosstalk with adipocytes has not been elucidated. Exosomes isolated from PC cell culture supernatant through ultracentrifugation were rich in long intergenic non-coding ROR (linc-ROR). After constructing PC cell lines with stable linc-ROR knockdown or overexpression via the transfection of short hairpin RNA (shRNA) and pLent-U6-GFP-Puro, direct and indirect coculture systems were established to simulate the interaction between adipocytes and PC cells. Next, the effects of conditioned medium collected from dedifferentiated adipocytes on PC cell proliferation, motility, metastasis, and epithelial-mesenchymal transition (EMT) were evaluated by western blot analysis, colony forming, real-time cell analysis (RTCA), 5-ethynyl-2'-deoxyuridine (EdU), immunofluorescence (IF), Transwell, and wound-healing assays in vitro. Xenograft models were employed to identify whether conditioned medium loaded with interleukin-1β (IL-1β) promoted PC cell growth in vivo. Our results demonstrate that linc-ROR delivery via exosomes represents a brand-new perspective of dedifferentiating adipocytes in the TME of PC, which further induce PC cell EMT via the hypoxia inducible factor 1α (HIF1α)-ZEB1 axis. Moreover, exosomal linc-ROR may become a novel diagnostic marker for PC patients.

Keywords: HIF1α; IL-1β; ZEB1; adipocytes; coculture system; epithelial-mesenchymal transition; exosomes; linc-ROR; pancreatic cancer.