The in vitro effect of norcantharidin on proliferation and invasion of human gallbladder carcinoma GBC-SD cells and its mechanism
Fan, Y-zu.; Fu, J-ye.; Zhao, Z-ming.; Chen, C-qiu.
Zhonghua Zhong Liu Za Zhi 26(5): 271-274
2004
ISSN/ISBN: 0253-3766 PMID: 15312360 Document Number: 584093
Objective To study the effect and mechanism of action of norcantharidin on proliferation and invasion of GBC-SD cells. Methods GBC-SD cells of human gallbladder carcinoma were cultured by cell culture technique. The tetrazolium-based colorimetric assay was used to evaluate cell growth. The Matrigel experiment and the crossing-river test were used to examine the invasiveness of GBC-SD cells. Expression of MMP2, TIMP2, PCNA and Ki-67 proteins of GBC-SD cells was determined by streptavidin-biotin complex method. Results Norcantharidin inhibited the growth and proliferation of GBC-SD cells in a dose and time dependent manner, with an IC50 value of 56.18 mug/ml at 48 h. The Matrigel experiment showed that norcantharidin began to inhibit the in vitro invasion of GBC-SD cells at the concentration of 5 mug/ml. At 40 g/ml, the invasive action of GBC-SD cells was inhibited completely and their crossing-river time was prolonged significantly. After treatment with norcantharidin, the expression of PCNA, Ki-67, 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 in vitro proliferation and growth of human gallbladder carcinoma cells at relatively low concentrations by inhibiting PCNA and Ki-67 expression. Its anti-invasive activity may be the results of decrease in MMP2 to TIMP2 ratio and reduced cell motility.