Brief Article
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World J Gastroenterol. Jun 28, 2011; 17(24): 2965-2971
Published online Jun 28, 2011. doi: 10.3748/wjg.v17.i24.2965
CD133+ gallbladder carcinoma cells exhibit self-renewal ability and tumorigenicity
Cheng-Jian Shi, Jun Gao, Min Wang, Xin Wang, Rui Tian, Feng Zhu, Ming Shen, Ren-Yi Qin
Cheng-Jian Shi, Min Wang, Xin Wang, Rui Tian, Feng Zhu, Ming Shen, Ren-Yi Qin, Department of Biliary-pancreatic Surgery, Affiliated Tongji Hospital, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, Hubei Province, China
Jun Gao, Department of General Surgery, The Affiliated Hospital of Qingdao University Medical College, Qingdao 266003, Shandong Province, China
Author contributions: Shi CJ and Gao J contributed equally to this work; Shi CJ, Gao J and Wang X performed the majority of the experiments, analyzed the data and wrote the manuscript; Tian R and Qin RY designed the research and drafted the manuscript; Wang X, Wang M and Shen M helped edit the manuscript; Zhu F contributed to the cell culture experiments.
Supported by Grant from the National Natural Science Foundation of China, No. 30772172
Correspondence to: Ren-Yi Qin, MD, Professor, Department of Biliary-pancreatic Surgery, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, No. 1095, Jiefang Ave., Wuhan 430030, Hubei Province, China. ryqin@tjh.tjmu.edu.cn
Telephone: +86-27-83663814 Fax: +86-27-83663874
Received: October 25, 2010
Revised: November 28, 2010
Accepted: December 5, 2010
Published online: June 28, 2011
Abstract

AIM: To identify cancer stem cells (CSCs) in human gallbladder carcinomas (GBCs).

METHODS: Primary GBC cells were cultured under serum-free conditions to produce floating spheres. The stem-cell properties of the sphere-forming cells, including self-renewal, differentiation potential, chemoresistance and tumorigenicity, were determined in vitro or in vivo. Cell surface expression of CD133 was investigated in primary tumors and in spheroid cells using flow cytometry. The sphere-colony-formation ability and tumorigenicity of CD133+ cells were assayed.

RESULTS: In vitro culture experiments revealed that floating spheroids were generated from primary GBC cells, and these sphere-forming cells could generate new progeny spheroids in serum-free media. Spheroid cells were differentiated under serum-containing conditions with downregulation of the stem cell markers Oct-4, Nanog, and nestin (P < 0.05). The differentiated cells showed lower spheroid-colony-formation ability than the original spheroid cells (P < 0.05). Spheroid cells were more resistant to chemotherapeutic reagents than the congenetic adherent cells (P < 0.05). Flow cytometry showed enriched CD133+ population in sphere-forming cells (P < 0.05). CD133+ cells possessed high colony-formation ability than the CD133- population (P < 0.01). CD133+ cells injected into nude mice revealed higher tumorigenicity than their antigen-negative counterparts (P < 0.05).

CONCLUSION: CD133 may be a cell surface marker for CSCs in GBC.

Keywords: Gallbladder carcinoma, Cancer stem cell, Non-adherent spheres, CD133 protein, Self-renewal, Tumorigenicity