Research Paper Volume 12, Issue 22 pp 22700—22718

Drug-selected population in melanoma A2058 cells as melanoma stem-like cells retained angiogenic features – the potential roles of heparan-sulfate binding ANGPTL4 protein

Chia-Yu Shih1, *, , Yu-Che Cheng1,2,3, *, , ChiaoHui Hsieh1, , TingTing Tseng1, , ShihSheng Jiang4, , Shao-Chen Lee1, ,

  • 1 School of Medicine, College of Medicine, Fu Jen Catholic University, New Taipei, Taiwan
  • 2 Proteomics Laboratory, Cathay Medical Research Institute, Cathay General Hospital, Taipei, Taiwan
  • 3 Department of Biomedical Science and Engineering, National Central University, Jhongli, Taiwan
  • 4 National Institute of Cancer Research, National Health Research Institutes, Miaoli, Taiwan
* Equal contribution

Received: September 5, 2019       Accepted: July 16, 2020       Published: November 10, 2020
How to Cite

Copyright: © 2020 Shih et al. This is an open access article distributed under the terms of the Creative Commons Attribution License (CC BY 3.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.


Malignant cancer may contain highly heterogeneous populations of cells, including stem-like cells which were resistant to chemotherapy agents, radiation, mechanical stress, and immune surveillance. The characterization of these specific subpopulations might be critical to develop novel strategy to remove malignant tumors.

We selected and enriched small population of human melanoma A2058 cells by repetitive selection cycles (selection, restoration, and amplification). These subpopulation of melanoma cells persisted the characteristics of slower cell proliferation, enhanced drug-resistance, elevated percentage of side population as analyzed by Hoechst33342 exclusion, in vitro sphere formation, and in vivo xenograft tumor formation by small amount of tumor cells. The selected populations would be melanoma stem-like cells with high expression of stem cell markers and altered kinase activation. Microarray and bioinformatics analysis highlighted the high expression of angiopoietin-like 4 protein in drug-selected melanoma stem-like cells. Further validation by specific shRNA demonstrated the role of angiopoietin-like 4 protein in drug-selected subpopulation associated with enhanced drug-resistance, sphere formation, reduced kinase activation, in vitro tube-forming ability correlated with heparan-sulfate proteoglycans.

Our finding would be applicable to explore the mechanism of melanoma stemness and use angiopoietin-like 4 as potential biomarkers to identify melanoma stem-like cells.


CSCs: cancer stem cells; ANGPTL4: angiopoietin-like protein4; FGFR3: fibroblast growth factor receptor 3; SDC1: syndecan-1; NDST1: N-deacetylase/N-sulfotransferase 1; HS2ST1: heparan sulfate 2-O-sulfotransferase 1; GEO: Gene Expression Omnibus.