有哪些信誉好的足球投注网站Sphere-forming cell subpopulations with cancer stem cell properties in human hepatoma cell lines.pdf
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Sphere-forming cell subpopulations with cancer stem cell properties in human hepatoma cell lines
RESEARCH ARTICLE Open Access
Sphere-forming cell subpopulations with cancer
stem cell properties in human hepatoma cell lines
Lu Cao1, Yanming Zhou2, Beibei Zhai1, Jian Liao1, Wen Xu1, Ruixiu Zhang1, Jing Li1, Yu Zhang1, Lei Chen1,
Haihua Qian1, Mengchao Wu1 and Zhengfeng Yin1*
Abstract
Background: Cancer stem cells (CSCs) are regarded as the cause of tumor formation and recurrence. The isolation
and identification of CSCs could help to develop novel therapeutic strategies specifically targeting CSCs.
Methods: Human hepatoma cell lines were plated in stem cell conditioned culture system allowed for sphere
forming. To evaluate the stemness characteristics of spheres, the self-renewal, proliferation, chemoresistance,
tumorigenicity of the PLC/PRF/5 sphere-forming cells, and the expression levels of stem cell related proteins in the
PLC/PRF/5 sphere-forming cells were assessed, comparing with the parental cells. The stem cell RT-PCR array was
performed to further explore the biological properties of liver CSCs.
Results: The PLC/PRF/5, MHCC97H and HepG2 cells could form clonal nonadherent 3-D spheres and be serially
passaged. The PLC/PRF/5 sphere-forming cells possessed a key criteria that define CSCs: persistent self-renewal,
extensive proliferation, drug resistance, overexpression of liver CSCs related proteins (Oct3/4, OV6, EpCAM, CD133
and CD44). Even 500 sphere-forming cells were able to form tumors in NOD/SCID mice, and the tumor initiating
capability was not decreased when spheres were passaged. Besides, downstream proteins DTX1 and Ep300 of the
CSL (CBF1 in humans, Suppressor of hairless in Drosophila and LAG1 in C. elegans) -independent Notch signaling
pathway were highly expressed in the spheres, and a gamma-secretase inhibitor MRK003 could significantly inhibit
the sphere formation ability.
Conclusions: Nonadherent tumor spheres from hepatoma cell lines cultured in stem cell conditioned medium
possess liver CSC properties, and the CSL-independent Notch signal
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