Abstract: Objective To evaluate inhibitive effect of norcantharidin on vasculogenic mimicry in three-dimensional cultures of gallbladder carcinoma cell lines GBC-SD and SGC-996 in vitro.Methods The different invasive capacity of gallbladder carcinoma cell lines GBC-SD and SGC-996 were detected by Transwell chamber and collagen gel contraction in vitro. Gallbladder carcinoma cell lines GBC-SD and SGC-996 were tested for their ability to form vasculogenic-like networks in three-dimensional cultures containing Matrigel and type Ⅰcollagen.Norcantharidin was dropped into three-dimensional culture model.The structures of vasculogenic-like networks were observed with light and electron microscope in different periods of time.Results ① The gallbladder carcinoma cell lines GBC-SD had higher invasion than SGC-996 (P＝0.0013),the aggressive gallbladder carcinoma cell lines GBC-SD were able to form in vitro vasculogenic-like networks in three-dimensional cultures containing Matrigel and type Ⅰcollagen,whereas the poorly aggressive SGC-996 cell lines were not.② From 1st day to 4th day the collagen contraction index of GBC-SD group containing TIMP2 were significantly lower than that of GBC-SD group (F=353.88,P<0.0001; F=525.80,P<0.0001; F=190.02,P=0.0002; F=196.26,P=0.0002),furthermore.The collagen contraction index of aggressive GBC-SD cell lines was higher than GBC-SD group poorly aggressive SGC-996 cell lines (F=318.73,P<0.0001; F=911.25,P<0.0001; F=268.88,P<0.0001; F=243.14,P<0.0001); the collagen contraction index of GBC-SD group containing TIMP1 was lower than that of GBC-SD group only at the second day (F=52.66,P=0.0019).③ the network formation could be blocked by Norcantharidin by inhibiting cell invasion and promoting cell apoptosis.Conclusion Three-dimensional cultures model can perfectly stimulate tumor cell growing and progressing conditions in vivo,the aggressive GBC-SD cell lines have higher invasive capacity than SGC-996 cell lines and can form in vitro vasculogenic-like networks in three-dimensional cultures containing Matrigel and type Ⅰcollagen,Norcantharidin can completely block the vasculogenic-like networks.

Key words: Gallbladder neoplasm, Vasculogenic mimicry, Norcantharidin