Cancer-associated fibroblasts enhance the chemoresistance of CD73+ hepatocellular carcinoma cancer cells via HGF-Met-ERK1/2 pathway

Hao Peng; Rong Xue; Zheng Ju; Jiannan Qiu; Jiawei Wang; Wei Yan; Xiaojie Gan; Yizhu Tian; Hongbin Shen; Xiaoming Wang; Xuehao Wang; Xuhao Ni; Yue Yu; Ling Lu

doi:10.21037/atm-20-1038

Cancer-associated fibroblasts enhance the chemoresistance of CD73⁺ hepatocellular carcinoma cancer cells via HGF-Met-ERK1/2 pathway

Ann Transl Med. 2020 Jul;8(14):856. doi: 10.21037/atm-20-1038.

Authors

Hao Peng¹, Rong Xue^{1

2}, Zheng Ju¹, Jiannan Qiu¹, Jiawei Wang¹, Wei Yan¹, Xiaojie Gan¹, Yizhu Tian¹, Hongbin Shen³, Xiaoming Wang⁴, Xuehao Wang¹, Xuhao Ni¹, Yue Yu¹, Ling Lu^{1

5

6}

Affiliations

¹ Hepatobiliary Center, The First Affiliated Hospital of Nanjing Medical University, Research Unit of Liver Transplantation and Transplant Immunology, Chinese Academy of Medical Sciences, Nanjing, China.
² School of Medicine, Southeast University, Nanjing, China.
³ Department of Epidemiology and Biostatistics, School of Public Health, Collaborative Innovation Center for Cancer Medicine, Nanjing Medical University, Nanjing, China.
⁴ State Key Laboratory of Reproductive Medicine, Department of Immunology, Nanjing Medical University, Nanjing, China.
⁵ Jiangsu Collaborative Innovation Center of Biomedical Functional Materials, College of Chemistry and Materials Science, Nanjing Normal University, Nanjing, China.
⁶ Jiangsu Key Laboratory of Cancer Biomarkers, Prevention and Treatment, Collaborative Innovation Center for Personalized Cancer Medicine, Nanjing Medical University, Nanjing, China.

Abstract

Background: Cancer-associated fibroblasts (CAFs) are a major component of hepatocellular carcinoma (HCC) stroma that are critically involved in HCC cancer chemoresistance, but the mechanism has not been elucidated. Previous studies have reported CD73 exerted an immunosuppressive function in cancer. Here, we explored the mechanism by which CAFs regulates CD73⁺ HCC cells and clarified whether CAFs promote chemoresistance of CD73⁺ cells.

Methods: We used the co-culture method to study the relationship between CAFs and HCC cells. Immunohistochemistry was applied to evaluate the correlation between α-smooth-muscle actin (α-SMA) and CD73. CD73 mRNA and protein were determined by real-time polymerase chain reaction (RT-PCR) and western blotting, and hepatocyte growth factor (HGF) was assayed by enzyme-linked immunosorbent assay (ELISA). Western blotting was used to explore the regulated pathway of CD73⁺ HCC. We then knocked down CD73 in cells, and then assessed the effect of CD73 on the apoptosis by flow cytometry. Finally, a sphere formation assay was applied to investigate the stemness of cancer cells, and xenograft tumors in nude mice were built to investigate the tumorigenicity.

Results: We found that the proportion of CAFs was positively correlated with CD73 expression in HCC cells. Mechanistically, c-Met and the MEK-ERK1/2 pathway were activated by HGF from CAFs which upregulated CD73 expression in HCC cells. Also, we found that CD73 promote sorafenib and cisplatin resistance in HCC, and CD73⁺ HCC cells indicated the higher capability of tumorigenicity compared to CD73^- HCC cells in vivo. Furthermore, HGF further enhanced the chemoresistant characteristics of CD73⁺ tumor cells.

Conclusions: Our findings collectively suggest that CD73 is a vital HCC-chemoresistance force controlled by cross-talking between CAFs and HCC cells, thereby establishing CD73 as a potential new therapeutic target for HCC.

Keywords: CD73; Hepatocellular carcinoma (HCC); cancer-associated fibroblast (CAF); chemoresistance.