CD133⁺EpCAM⁺ Phenotype Possesses More Characteristics of Tumor Initiating Cells in Hepatocellular Carcinoma Huh7 Cells

Abstract

Background: EpCAM or CD133 has been used as the tumor initiating cells (TICs) marker in hepatocellular carcinoma (HCC). We investigated whether cells expressing with both EpCAM and CD133 surface marker were more representative for TICs in hepatocellular carcinoma Huh7 cells.Methods: Four different phenotypes of CD133+EpCAM+, CD133+EpCAM-, CD133-EpCAM+ and CD133-EpCAM- in Huh7 cells were sorted by flow cytometry. Then cell differentiation, self-renewal, drug-resistance, spheroid formation and the levels of stem cell-related genes were detected to compare the characteristics of TICs. The ability of tumorigenicity was measured in nonobese diabetic/severe combined immunodeficient (NOD/SCID) mice to verify TICs.Results: CD133+EpCAM+ cells have many characteristics of TICs in Huh7 cells compared with CD133+EpCAM-, CD133-EpCAM+, CD133-EpCAM- cells, including enrichment in side population cells, higher differentiation capacity, increased colony-formation ability, preferential expression of stem cell-related genes, appearance of drug-resistant to some chemotherapeutics, more spheroid formation of culture cells and stronger tumorigenicity in NOD/SCID mice.Conclusion: CD133+EpCAM+ phenotype is precisely represented TICs in Huh7 cells. It might be useful for studying biology mechanism of TICs in hepatocellular carcinoma and screening new targets for cancer therapy.

Preclinical Evaluation of Transcriptional Targeting Strategy for Human Hepatocellular Carcinoma in an Orthotopic Xenograft Mouse Model

“…Prolonged 5-FU treatment has been shown to increase in EpCAM high and CD133 high 5-FU-resistant hepatocellular carcinoma cells (HuH-7; ref17). Hepatocellular carcinoma cells expressing EpCAM and CD133 proteins have been implicated to possess features characteristics of tumor-initiating cells (50). Interestingly, EpCAM high and CD133 high 5-FU-resistant hepatocellular carcinoma cells could be selectively killed by DZNep (17).…”

Surface markers of hepatocellular cancer stem cells and their clinical potential

“…Moreover, these cells were capable of initiating highly invasive HCC in nonobese diabetic, severe combined immunodeficient mice. Chen et al [77] found that CD133(+) EpCAM (+) cells possessed many characteristics of TICs in Huh-7 cells, including higher differentiation capacity, increased colony-formation ability, preferential expression of stem cell-related genes, drug-resistant to some chemothera-peutics, more spheroid formation of culture cells and stronger tumorigenicity in NOD/SCID mice. In 2013, Yamashita et al [78] showed that CSC markers EpCAM and CD90 was independently expressed in liver cancer.…”

Lidamycin decreases CD133 expression in hepatocellular carcinoma via the Notch signaling pathway

“…Cancer stem cells (CSCs) have been identified in a number of solid tumors, including HCC (2)(3)(4). CSCs may lead to recurrence, drug-resistance and tumor formation in HCC (5).…”

“…CSCs may lead to recurrence, drug-resistance and tumor formation in HCC (5). Cluster of differentiation (CD)133 is a membrane-bound pentaspan glycoprotein that has been identified as a surface biomarker in a variety of cancer types, including breast, colon, prostate, pancreatic, lung and liver carcinoma (2,3,(6)(7)(8)(9)(10)(11). Although its function remains unclear, directly targeting CD133 has been demonstrated to be a potentially effective strategy for eliminating CSCs (11,12).…”

“…To investigate this, the proportion of CD133 + Sphere formation assay. The sphere formation medium for HCC was created as previously described (2,15). Sorted CD133 + cells in sphere formation medium were cultured in ultra-low attachment 24-well plates (Corning Inc., Corning, NY, USA) at a density of 5,000 cells/well.…”