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CLC number: R73

On-line Access: 2011-04-06

Received: 2010-08-16

Revision Accepted: 2010-12-02

Crosschecked: 2011-03-11

Cited: 15

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Journal of Zhejiang University SCIENCE B 2011 Vol.12 No.4 P.256-263

http://doi.org/10.1631/jzus.B1000303


Preliminary screening and identification of stem cell-like sphere clones in a gallbladder cancer cell line GBC-SD


Author(s):  Bao-bing Yin, Shuang-jie Wu, Hua-jie Zong, Bao-jin Ma, Duan Cai

Affiliation(s):  Department of General Surgery, Huashan Hospital, Fudan University, Shanghai 200040, China

Corresponding email(s):   yinbaobing@hotmail.com, mcswq@yahoo.com.cn

Key Words:  Gallbladder cancer, Stem cell gene, Sphere clone, Suspension culture


Bao-bing Yin, Shuang-jie Wu, Hua-jie Zong, Bao-jin Ma, Duan Cai. Preliminary screening and identification of stem cell-like sphere clones in a gallbladder cancer cell line GBC-SD[J]. Journal of Zhejiang University Science B, 2011, 12(4): 256-263.

@article{title="Preliminary screening and identification of stem cell-like sphere clones in a gallbladder cancer cell line GBC-SD",
author="Bao-bing Yin, Shuang-jie Wu, Hua-jie Zong, Bao-jin Ma, Duan Cai",
journal="Journal of Zhejiang University Science B",
volume="12",
number="4",
pages="256-263",
year="2011",
publisher="Zhejiang University Press & Springer",
doi="10.1631/jzus.B1000303"
}

%0 Journal Article
%T Preliminary screening and identification of stem cell-like sphere clones in a gallbladder cancer cell line GBC-SD
%A Bao-bing Yin
%A Shuang-jie Wu
%A Hua-jie Zong
%A Bao-jin Ma
%A Duan Cai
%J Journal of Zhejiang University SCIENCE B
%V 12
%N 4
%P 256-263
%@ 1673-1581
%D 2011
%I Zhejiang University Press & Springer
%DOI 10.1631/jzus.B1000303

TY - JOUR
T1 - Preliminary screening and identification of stem cell-like sphere clones in a gallbladder cancer cell line GBC-SD
A1 - Bao-bing Yin
A1 - Shuang-jie Wu
A1 - Hua-jie Zong
A1 - Bao-jin Ma
A1 - Duan Cai
J0 - Journal of Zhejiang University Science B
VL - 12
IS - 4
SP - 256
EP - 263
%@ 1673-1581
Y1 - 2011
PB - Zhejiang University Press & Springer
ER -
DOI - 10.1631/jzus.B1000303


Abstract: 
This paper aims to screen and identify sphere clone cells with characteristics similar to cancer stem cells in human gallbladder cancer cell line GBC-SD. GBC-SD cells were cultured in a serum-free culture medium with different concentrations of the chemotherapeutic drug cisplatin for generating sphere clones. The mRNA expressions of stem cell-related genes CD133, OCT-4, Nanog, and drug resistance genes ABCG2 and MDR-1 in sphere clones were detected by quantitative real-time polymerase chain reaction (PCR). Stem cell markers were also analyzed by flow cytometry and immunofluorescent staining. Different amounts of sphere clones were injected into nude mice to test their abilities to form tumors. sphere clones were formed in serum-free culture medium containing cisplatin (30 μmol/L). Flow cytometry results demonstrated that the sphere clones expressed high levels of stem cell markers CD133+ (97.6%) and CD44+ (77.9%) and low levels of CD24+ (2.3%). These clones also overexpressed the drug resistance genes ABCG2 and MDR-1. Quantitative real-time PCR showed that sphere clones expressed stem cell genes Nanog and OCT-4 284 and 266 times, respectively, more than those in the original GBC-SD cells. Immunofluorescent staining showed that sphere clones overexpressed OCT-4, Nanog, and SOX-2, and low expressed MUC1 and vimentin. Tumor formation experiments showed that 1×103 sphere clone cells could induce much larger tumors in nude mice than 1×105 GBC-SD cells. In conclusion, sphere clones of gallbladder cancer with stem cell-like characteristics can be obtained using suspension cultures of GBC-SD cells in serum-free culture medium containing cisplatin.

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