Effect of norcantharidin on proliferation and invasion of human gallbladder carcinoma GBC-SD cells.
OBJECTIVE
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 microg/mL at 48 h. Norcantharidin began to inhibit the invasion of GBC-SD cells at the concentration of 5 microg/mL, and the invasive action of GBC-SD cells was inhibited completely and their crossing-river time was prolonged significantly at 40 microg/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).


CONCLUSIONS
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.
Fan
Yue-Zu
YZ
Department of General Surgery, Tongji Hospital of Tongji University, 389 Xincun Road, Shanghai 200065, China. fanyuezu_shtj@msn.com.
Fu
Jin-Ye
JY
Zhao
Ze-Ming
ZM
Chen
Cun-Qiu
CQ
D019086:Bridged Bicyclo Compounds, Heterocyclic / Q000494:pharmacology*; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects; D003094:Collagen; D004338:Drug Combinations; D005706:Gallbladder Neoplasms / Q000188:drug therapy* / Q000378:metabolism / Q000473:pathology*; D006801:Humans; D066298:In Vitro Techniques; D019394:Ki-67 Antigen / Q000378:metabolism; D007797:Laminin; D020778:Matrix Metalloproteinase 2 / Q000378:metabolism; D008516:Medicine, Chinese Traditional; D009361:Neoplasm Invasiveness; D018809:Proliferating Cell Nuclear Antigen / Q000378:metabolism; D011509:Proteoglycans; D019716:Tissue Inhibitor of Metalloproteinase-2 / Q000378:metabolism
D019086:Bridged Bicyclo Compounds, Heterocyclic; D004338:Drug Combinations; D019394:Ki-67 Antigen; D007797:Laminin; D018809:Proliferating Cell Nuclear Antigen; D011509:Proteoglycans; C059485:matrigel; D019716:Tissue Inhibitor of Metalloproteinase-2; C069741:norcantharidin; D003094:Collagen; D020778:Matrix Metalloproteinase 2