Acidic Microenvironment Up-Regulates Exosomal miR-21 and miR-10b in Early-Stage Hepatocellular Carcinoma to Promote Cancer Cell Proliferation and Metastasis
Authors: Chen-Yuan Wang
Feng-Wei Wang
Xiao-Han Jin
Rui-Hua Xu
Xiao-Peng Tian
Qing-Qing Cai
Jing-Ping Yun
Wei-Juan Huang
Mei Li
Dan Xie
Journal: Theranostics (2019)
Discipline: CLINICAL MEDICINE
Keywords: acidic microenvironment; early-stage hepatocellular carcinoma; epithelial- mesenchymal transition; miR-10b; miR-21.
Affiliations: Sun Yat-sen University Cancer Center, State Key Laboratory of Oncology in South China, Collaborative Innovation Center of Cancer Medicine, Guangzhou, China., Department of Reproductive Medicine, The Second Affiliated Hospital of Guangzhou University of Chinese Medicine, Guangzhou, China., Department of Pathology, Sun Yat-sen University Cancer Center, Guangzhou, China., Department of Pharmacology, College of Pharmacy, Jinan University, Guangzhou, China., Department of Medical Oncology, Sun Yat-sen University Cancer Center, Guangzhou, China.

Abstract:  Rationale: The incidence of hepatocellular carcinoma is rising worldwide. It is predicted that nearly half of the early-stage hepatocellular carcinoma (E-HCC) patients will develop recurrence. Dysregulated pH, a hallmark of E-HCC, is correlated with poor prognosis. The acidic microenvironment has been shown to promote the release of exosomes, the membrane vesicles recognized as intercellular communicators associated with tumor progression, recurrence, and metastasis. We, therefore, aimed to identify exosomes induced by acidic microenvironment that may regulate E-HCC progression and to explore their mechanisms and clinical significance in E-HCCs. Methods: miRNA microarray analysis and LASSO logistic statistic model were used to identify the main functional exosomal miRNAs. Invasion and scratch assays were performed to examine the migration and invasion of HCC cells. Immunoblotting and immunofluorescence were employed to detect the epithelial-to-mesenchymal transition (EMT) in HCC cells. Chromatin immunoprecipitation (ChIP) was used to analyze the binding of HIF-1Œ± and HIF-2Œ± to promoter regions of miR-21 and miR-10b. Results: The acidic microenvironment in HCC was correlated with poor prognosis of patients. Exosomes from HCC cells cultured in the acidic medium could promote cell proliferation, migration, and invasion of recipient HCC cells. We identified miR-21 and miR-10b as the most important functional miRNAs in acidic HCC-derived exosomes. Also, the acidic microenvironment triggered the activation of HIF-1Œ± and HIF-2Œ± and stimulated exosomal miR-21 and miR-10b expression substantially promoting HCC cell proliferation, migration, and invasion both in vivo and in vitro. In E-HCC patients, serum exosomal miR-21 and miR-10b levels were associated with advanced tumor stage and HIF-1Œ± and HIF-2Œ± expression and were independent prognostic factors for disease-free survival of E-HCC patients. Most importantly, we developed a nano-drug to target exosomal miR-21 and/or miR-10b and examined its therapeutic effects against HCC in vivo. Conclusion: Our findings suggested that the exosomal miR-21 and miR-10b induced by acidic microenvironment in HCC promote cancer cell proliferation and metastasis and may serve as prognostic molecular markers and therapeutic targets for HCC. 

Retracted: No

PubPeer comments:
 This seminal paper reports a collaborative effort between the Aguzzi lab and Nobel Prize winner Kurt Wu00fctrich. Together, they showed unequivocally that quasi-physiological expression levels of a prion protein variant results in spontaneous transmissible disease (see Kaplan-Meyer curve, Fig. 1B). This recapitulates the human situation in which familial Creutzfeldt-Jakob Disease is autosomal-dominant, but at the same time is infectious. The structure of this particular mutant is very well understood in atomistic detail. The  rigid loop of PrP thus emerges as a central determinant of prion infectivity.  ,