Identification of side population cells in human hepatocellular carcinoma cell lines with stepwise metastatic potentials.
OBJECTIVE
To identify the side population (SP) cells from four hepatocellular carcinoma (HCC) cell lines with stepwise metastatic potentials.


METHODS
SP cells were sorted from HCCLM3, MHCC97-H, MHCC97-L and Hep3B by flow cytometry, and then analyzed by differentiation study, clonogenic assay, chemoresistance study and tumorigenicity assay in vivo. The expression of ABCG(2) in SP cells was detected by immunocytochemistry, western blotting and real-time quantitative PCR, respectively.


RESULTS
There was significant difference in SP proportion among HCCLM3, MHCC97-H, MHCC97-L and Hep3B (28.7 +/- 1.6%, 14.5 +/- 0.6%, 4.2 +/- 0.4%, 0.9 +/- 0.1%, respectively, P < 0.01). All the SP cells showed similar characteristics of self-renewal, high clonogenicity, remarkable chemo-resistance and high expression of ABCG(2). As low as 2,000 SP cells could initiate tumors in non-obese diabetic/severe combined immunodeficiency mice successfully.


CONCLUSIONS
SP cells purified from HCC cell lines harbors cancer stem cell-like properties, and may be related to the metastatic potentials and therapeutic-resistance of HCC.
Shi
Guo-Ming
GM
Key Laboratory for Carcinogenesis and Cancer Invasion, Liver Cancer Institute, Zhongshan Hospital and Shanghai Medical School, Fudan University, the Chinese Ministry of Education, 136 Yi Xue Yuan Road, Shanghai, 200032, People's Republic of China.
Xu
Yang
Y
Fan
Jia
J
Zhou
Jian
J
Yang
Xin-Rong
XR
Qiu
Shuang-Jian
SJ
Liao
Yong
Y
Wu
Wei-Zhong
WZ
Ji
Yuan
Y
Ke
Ai-Wu
AW
Ding
Zhen-Bin
ZB
He
Yi-Zhou
YZ
Wu
Bing
B
Yang
Guo-Huan
GH
Qin
Wen-Zhen
WZ
Zhang
Wu
W
Zhu
Jiang
J
Min
Zhi-Hui
ZH
Wu
Zhi-Quan
ZQ
D000070997:ATP Binding Cassette Transporter, Subfamily G, Member 2; D018528:ATP-Binding Cassette Transporters / Q000235:genetics; D006528:Carcinoma, Hepatocellular / Q000145:classification* / Q000453:epidemiology / Q000401:mortality / Q000473:pathology*; D002454:Cell Differentiation; D002455:Cell Division; D045744:Cell Line, Tumor; D019008:Drug Resistance, Neoplasm; D005434:Flow Cytometry; D015972:Gene Expression Regulation, Neoplastic; D006801:Humans; D008113:Liver Neoplasms / Q000145:classification* / Q000453:epidemiology / Q000401:mortality / Q000473:pathology*; D009362:Neoplasm Metastasis / Q000473:pathology*; D009363:Neoplasm Proteins / Q000235:genetics; D010641:Phenotype; D011379:Prognosis; D015996:Survival Rate; D014410:Tumor Stem Cell Assay
C116614:ABCG2 protein, human; D000070997:ATP Binding Cassette Transporter, Subfamily G, Member 2; D018528:ATP-Binding Cassette Transporters; D009363:Neoplasm Proteins