目的 探讨去甲斑蝥素对GBC-SD、SGC-996胆囊癌细胞系体外三维培养血管生成拟态形态学影响。方法 Transwell小室及鼠尾Ⅰ型胶原收缩试验测定GBC-SD、SGC-996胆囊癌细胞系体外侵袭迁徙力, 运用Matrigel胶、鼠尾Ⅰ型胶原三维培养基质建立GBC-SD、SGC-996胆囊癌细胞三维培养模型, 观察细胞侵袭力高低与血管生成拟态相关性, 并运用电镜、光镜观察血管生成拟态形态学构成;在三维培养模型中加入NCTD后动态观察NCTD对血管生成拟态形态影响。结果 ①GBC-SD胆囊癌细胞侵袭迁徙能力显著高于SGC-996胆囊癌细胞(P＝0.0013), 高侵袭GBC-SD胆囊癌细胞在Matrigel胶、鼠尾Ⅰ型胶原三维培养基质中均能形成管状结构, 而低侵袭SGC-996胆囊癌细胞不能形成管状结构 ②第1天至第4天, GBC-SD TIMP2干预组收缩指数明显低于GBC-SD组(F=353.88, P<0.0001;F=525.8, P<0.0001;F=190.02, P=0.0002;F=196.26, P=0.0002), 高侵袭性GBC-SD细胞收缩指数显著高于低侵袭性SGC-996细胞(F=318.73, P<0.0001;F=911.25, P<0.0001;F=268.88, P<0.0001;F=243.14, P<0.0001);仅第2天, GBC-SD TIMP1干预组收缩指数显著低于GBC-SD组(F=52.66, P=0.0019)③NCTD可抑制胆囊癌细胞侵袭迁移能力, 促使细胞凋亡, 抑制细胞三维培养基质中管道形成。结论 三维培养模型能模拟体内细胞生长条件, GBC-SD胆囊癌细胞侵袭迁徙能力显著高于SGC-996胆囊癌细胞并能在三维培养基质中形成管状结构, NCTD能抑制胆囊癌细胞体外血管生成拟态形成。

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.