Low expression of SPARC in gastric cancer-associated fibroblasts leads to stemness transformation and 5-fluorouracil resistance in gastric cancer

Yongchen Ma; Jing Zhu; Shanwen Chen; Ju Ma; Xiaoqian Zhang; Sixia Huang; Jianwen Hu; Taohua Yue; Junling Zhang; Pengyuan Wang; Xin Wang; Long Rong; Hongjie Guo; Guowei Chen; Yucun Liu

doi:10.1186/s12935-019-0844-8

Low expression of SPARC in gastric cancer-associated fibroblasts leads to stemness transformation and 5-fluorouracil resistance in gastric cancer

Cancer Cell Int. 2019 May 21:19:137. doi: 10.1186/s12935-019-0844-8. eCollection 2019.

Authors

Yongchen Ma¹, Jing Zhu¹, Shanwen Chen¹, Ju Ma¹, Xiaoqian Zhang¹, Sixia Huang², Jianwen Hu¹, Taohua Yue¹, Junling Zhang¹, Pengyuan Wang¹, Xin Wang¹, Long Rong³, Hongjie Guo⁴, Guowei Chen¹, Yucun Liu¹

Affiliations

¹ 1Department of General Surgery, Peking University First Hospital, Beijing, 100034 People's Republic of China.
² 2Department of Pathology, Peking University First Hospital, Beijing, 100034 People's Republic of China.
³ 3Department of Endoscopic Center, Peking University First Hospital, Beijing, 100034 People's Republic of China.
⁴ 4Department of Interventional Radiology and Vascular Surgery, Peking University First Hospital, Beijing, 100034 People's Republic of China.

Abstract

Background: The aim of the present study was to clarify the correlations between SPARC expression in gastric cancer-associated fibroblasts (GCAFs) and the prognosis of patients with gastric cancer and to elucidate the role of GCAF-derived SPARC in stemness transformation and 5-fluorouracil resistance in gastric cancer.

Methods: One hundred ninety-two patients were enrolled in the present study. SPARC expression levels were evaluated by immunohistochemical staining. Primary GCAFs were obtained and cultured from cancer patients for in vitro study, and a lentivirus infection method was employed to knock down SPARC expression in GCAFs. The stemness phenotype and 5-fluorouracil (5-FU) response of gastric cancer cells were assessed via a 3D co-culture model. The apoptotic status and stemness alterations were monitored by flow cytometry and western blotting. Additionally, label-free quantification proteomics was used to identify the differentially expressed proteins and potential pathways in gastric cancer cells treated with GCAF-derived SPARC.

Results: Low expression of GCAF-derived SPARC was associated with decreased differentiation and reduced 5-year overall survival and was an independent predictive factor for prognosis in gastric cancer. The 3D tumour growth and 5-FU resistance abilities of gastric cancer cells were elevated after treatment with GCAFs with SPARC knockdown relative to these abilities in negative control cells. Additionally, suppressing SPARC expression in GCAFs facilitated the phenotypic alteration of gastric cancer cells towards CD44⁺/CD24^- cancer stem cell (CSC)-like cells. Quantification proteomics analysis revealed that the differentially expressed proteins in gastric cancer cells were mainly involved in the AKT/mTOR and MEK/ERK signalling pathways.

Conclusions: SPARC expression in GCAFs is a useful prognostic factor in patients with gastric cancer. Low expression of GCAF-derived SPARC can lead to CSC transformation and 5-FU resistance. Additionally, the AKT/mTOR and MEK/ERK signalling pathways may participate in the malignant process.

Keywords: Cancer-associated fibroblasts; Drug resistance; Gastric cancer; SPARC; Stemness.