CD133+ subpopulation of the HT1080 human fibrosarcoma cell line exhibits cancer stem-like characteristics

Abstract

The cancer stem cell (CSC) theory holds that a minority population within tumors possesses stem cell properties of self-renewal and multilineage differentiation capacity and provides the initiating cells from which tumors are derived and sustained. However, verifying the existence of these CSCs has been a significant challenge. The CD133 antigen is a pentaspan membrane glycoprotein proposed to be a CSC marker for cancer-initiating subpopulations in the brain, colon and various other tissues. Here, CD133+ cells were obtained and characterized from the HT1080 cell line to determine the utility of this marker for isolating CSCs from human fibrosarcoma cells. In this study, CD133+ cells were separated from HT1080 cells using magnetic beads and characterized for their proliferation rate and resistance to chemotherapeutic drugs, cisplatin and doxorubicin, by MTS assay. Relative expression of tumor-associated genes Sox2, Oct3/4, Nanog, c-Myc, Bmi-1 and ABCG2 was measured by real-time polymerase chain reaction (PCR). Clonal sphere formation and the ability of CD133+ cells to initiate tumors in BALB/c nude mice was also evaluated. We found that CD133+ cells showed a high proliferation rate, increased resistance to chemotherapy drugs and overexpression of tumor-associated genes compared with these features in CD133- cells. Additionally, CD133+ cells were able to form spherical clusters in serum-free medium with high clonogenic efficiency, indicating a significantly greater tumor-initiating potential when compared with CD133- cells. These findings indicate that CD133+ cells identified within the HT1080 human fibrosarcoma cell line possess many CSC properties and may facilitate the development of improved therapies for fibrosarcoma.

Transition from mesenchymal to bleb‐based motility is predominantly exhibited by CD133‐positive subpopulation of fibrosarcoma cells

“…Both MACS-and FACS-isolated fractions of CD133+ HT1080 cells were significantly enriched with cells capable of MAT. These results are aligned with another study, which showed that the CD133+ HT1080 cells were able to form a greater number of tumours when injected into mice, possess multi-drug resistance properties and form spherical clusters in serum-free medium [Feng et al, 2013]. In our experiments, some CD133-negative cells were also able to switch to blebbing.…”

“…To test our hypothesis, we used a well-characterised stem cell marker, CD133, which has been first identified in neuronal stem cells [Mizrak et al, 2008] and then shown to be associated with CSCs in various tumours [Singh et al, 2003;Tirino et al, 2011;Han et al, 2015]. Notably, HT1080 fibrosarcoma cells have been shown to contain a subpopulation of CD133-positive cells that also satisfy other criteria of being CSCs, including the formation of clonal spheres in vitro, efficient tumorigenesis in mice, increased proliferation rate, resistance to chemotherapy and overexpression of tumour-associated genes [Feng et al, 2013]. To determine whether the fractions of CD133positive and blebbing cells overlap, we analysed protrusive activity of unsorted HT1080 cells in the presence of 200 µM CK-666 by live cell imaging, then fixed them and performed their post hoc CD133 staining.…”

“…CD133+ cells are also present in various sarcomas [Suva et al, 2009]. In particular, CD133 expression defines a minor subpopulation of HT1080 cells with enhanced stemness features [Tirino et al, 2011;Feng et al, 2013]. Taking into account that induction of MAT in melanoma cells positively correlates with the expression of genes characteristic for CSCs [Taddei et al, 2011[Taddei et al, , 2014, we hypothesised that the MAT-competent fibrosarcoma cells may correspond to CSCs.…”

Stat3/Oct-4/c-Myc signal circuit for regulating stemness-mediated doxorubicin resistance of triple-negative breast cancer cells and inhibitory effects of WP1066

“…ABCG2 is not only associated with MDR, but may also function as a breast cancer stem cell marker (20,34,35,37,38). Although CD133 was initially described as a specific marker of human hematopoietic stem cells (13,14,17,34,39,40), CD133 has been used as the primary marker of putative CSCs and has been reported to be the most reproducible marker of breast CSCs (BCSCs) (14,24,27,39,41). Therefore, cells expressing high levels of cell surface markers, including CD44, ABCG2 and CD133, have stem-like activities (10, 17,24).…”

Molecular radiosensitization of soft tissue sarcoma by oncolytic virus-mediated MCL1 ablation

“…The dormancy of such cells is a critical factor for the radioresistance of tumors, because radiotherapy targets proliferating cancer cells by inducing DNA damage-related cell cycle arrest. Recent reports have suggested that a CD133-positive subpopulation of radioresistant HT1080 cells exhibit cancer stem-like characteristics [36,37], such as activation of stemness-related markers, sphere-forming capacity, tumorigenicity, and resistance to chemotherapy [37]. CD133-positive subpopulations have similarly been associated with the radioresistance of brain tumors [38], liver cancer [39], and gastric cancer [40].…”