Basic Research
Copyright ©2005 Baishideng Publishing Group Inc. All rights reserved.
World J Gastroenterol. Apr 28, 2005; 11(16): 2431-2437
Published online Apr 28, 2005. doi: 10.3748/wjg.v11.i16.2431
Effect of norcantharidin on proliferation and invasion of human gallbladder carcinoma GBC-SD cells
Yue-Zu Fan, Jin-Ye Fu, Ze-Ming Zhao, Cun-Qiu Chen
Yue-Zu Fan, Ze-Ming Zhao, Cun-Qiu Chen, Department of General Surgery, Tongji Hospital of Tongji University, Shanghai 200065, China
Jin-Ye Fu, Department of Surgery, Pudong People Hospital, Shanghai 201200, China
Author contributions: All authors contributed equally to the work.
Supported by the Scientific Foundation of Tongji University, China, No. KPB027
Correspondence to: Professor, Director Yue-Zu Fan, Department of General Surgery, Tongji Hospital of Tongji University, 389 Xincun Road, Shanghai 200065, China. fanyuezu_shtj@msn.com
Telephone: +86-21-56051080-1107 Fax: +86-21-56050502
Received: July 28, 2004
Revised: July 29, 2004
Accepted: September 4, 2004
Published online: April 28, 2005
Abstract

AIM: To investigate the effect of norcantharidin on proliferation and invasion of human gallbladder carcinoma GBC-SD cells in vitro and its anticancer mechanism.

METHODS: Human gallbladder carcinoma GBC-SD cells were cultured by cell culture technique. The growth and the invasiveness of GBC-SD cells in vitro were evaluated by the tetrazolium-based colorimetric assay and by the Matrigel experiment and the crossing-river test. Expression of PCNA, Ki-67, MMP2 and TIMP2 proteins of GBC-SD cells was determined by streptavidin–biotin complex method.

RESULTS: In vitro norcantharidin inhibited the growth and proliferation of GBC-SD cells in a dose- and time-dependent manner, with the IC50 value of 56.18 μg/mL at 48 h. Norcantharidin began to inhibit the invasion of GBC-SD cells at the concentration of 5 μg/mL, and the invasive action of GBC-SD cells was inhibited completely and their crossing-river time was prolonged significantly at 40 μg/mL. After treatment with norcantharidin, the expression of PCNA, Ki-67, and MMP2 was significantly decreased. With the increase in TIMP2 expression, the MMP2 to TIMP2 ratio was decreased significantly (P<0.05).

CONCLUSION: Norcantharidin inhibits the proliferation and growth of human gallbladder carcinoma cells in vitro at relatively low concentrations by inhibiting PCNA and Ki-67 expression. Its anti-invasive activity may be the result of decrease in MMP2 to TIMP2 ratio and reduced cell motility.

Keywords: Norcantharidin, Gallbladder neoplasm, Cell culture, Proliferation, Invasion, Oncoprotein PCNA, Ki-67, MMP2 and TIMP2, Immunohistochemistry