Differential capacity of CD90+ cells in autophagy activation following chemotherapy in hepatocellular carcinoma.

Abstract

INTRODUCTION AND OBJECTIVES:

Analysis of cancer biomarkers is an important tool in developing targeted-therapy and in modulating chemoresistance. Here, we analyze the relevance of CD90, a marker of cancer stem cells (CSC) in hepatocellular carcinoma (HCC) and its correlation with autophagy.

MATERIALS AND METHODS:

For in vivo study, 86 specimens were collected from 43 patients undergoing liver resections. In each patient, HCC nodule (HCC) and surrounding non-tumor (SNT) were collected. For in vitro study, HCC cells JHH6 subpopulations expressing CD90+ and CD90- were isolated using magnetic-sorter and confirmed by flow-cytometry. Upon doxorubicin treatment, autophagy turn-over was analyzed by RTqPCR for mRNA expression, Western blot for protein expression, and autophagosome staining for autophagy-flux. Cytotoxicity test was performed by MTT assay. Gene and protein analysis were performed in clinical samples together with immunohistostaining.

RESULTS:

CD90 mRNA expression was higher in HCC than in SNT for 8-fold (p < 0.001). LC3-II protein was up-regulated in the HCC in comparison with the SNT (p < 0.05). In vitro model showed that CD90+ and CD90- cells had diverse expressions of autophagy-related genes. Upon doxorubicin treatment, autophagy was activated in both cells by increasing LC3-II protein expression, autophagic vacuoles, and dysregulation of autophagy-related mRNAs. A differential autophagic capacity was noticed between two subpopulations and it was correlated with cellular toxicity assay.

CONCLUSIONS:

We demonstrated the relevance of differential autophagy capacity of CD90+ cells in HCC. Autophagy was involved in cancer-defense mechanism against doxorubicin. Cancer promoting function of autophagy in CD90+ cells was also related to cancer environment.

• Authors: Huy Q Do, An B Luong, Deborah Bonazza, Cristina Bottin, Thao Pt Doan, Long Dc Tran, Nhung H Truong, Gianluca Tell, Hoa Lt Pham, Claudio Tiribelli, Caecilia Hc Sukowati
• Journal: Annals of hepatology (Ann Hepatol) 2020 Nov - Dec Vol. 19 Issue 6 Pg. 645-652 ISSN: 1665-2681 [Print] Mexico
• PMID: 32745631 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
• Copyright: Copyright © 2020 Fundación Clínica Médica Sur, A.C. Published by Elsevier España, S.L.U. All rights reserved.

Chemical References

• Antibiotics, Antineoplastic
• MAP1LC3B protein, human
• Microtubule-Associated Proteins
• Thy-1 Antigens
• Doxorubicin

Topics

• Antibiotics, Antineoplastic (therapeutic use)
• Autophagy (drug effects)
• Carcinoma, Hepatocellular (drug therapy, metabolism, pathology)
• Cell Culture Techniques
• Cell Line, Tumor
• Doxorubicin (therapeutic use)
• Female
• Humans
• Liver Neoplasms (drug therapy, metabolism, pathology)
• Male
• Microtubule-Associated Proteins (metabolism)
• Middle Aged
• Thy-1 Antigens (metabolism)