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.
Sun Wei
,
Fan Yue-Zu
,
Zhang Wen-Zhong
,
Ge Chun-Yan
,
Zhao Feng-Di
,
Zhou Ting-Ting
,
Ning Yan-Xia
. Inhibitive effect of norcantharidin on vasculogenic mimicry in three-dimensional cultures of gallbladder carcinoma cell lines GBC-SD and SGC-996 in vitro[J]. Surgical Research and New Technique, 2012
, 1(1)
: 42
-48
.
DOI: 10.3969/j.issn.2095-378X.2012.01.008
[1] Maniotis AJ,Folberg R,Hess A,et al.Vascular channel formation by human melanoma cells in vivo and in vitro:vasculogenic mimicry[J].Am J Pathol,1999,155(3):739-752.
[2] Shirakawa K,Wakasugi H,Heike Y,et al.Vasculogenic mimicry and pseudo-comedo formation in breast cancer[J].Int J Cancer,2002(6),99:821-828.
[3] 赵秀兰,杜静,张诗武,等.肝细胞癌中血管生成拟态的研究[J].中华肝脏病杂志,2006,14(1):41-44.
[4] 范跃祖,孙伟,张文忠,等.原发性胆囊癌患者肿瘤血管生成拟态及其临床意义[J].中华医学杂志,2007,87(3):145-149.
[5] Folberg R,Hendrix MJ,Maniotis AJ,et al.Vasculogenic mimicry and tumor angiogenesis[J].Am J Pathol,2000,156(2):361-381.
[6] Sood AK,Fletcher MS,Coffin JE,et al.Functional role of matrix metalloproteinases in ovarian tumor cell plasticity[J].Am J Obstet Gynecol,2004,190(4):899-909.
[7] Hess AR,Seftor EA,Seftor RE, et al.Phosphoinositide 3-kinase regulates membrane Type 1-matrix metalloproteinase (MMP) and MMP-2 activity during melanoma cell vasculogenic mimicry[J].Cancer Res,2003,63(16):4757-4762.
[8] 孙伟,范跃祖,张文忠,等.Ⅳ型胶原和基质金属蛋白酶-2、-9在胆囊癌血管生成拟态组织中的表达与意义[J].中华实验外科杂志,2007,24(2):1472-1474.
[9] 范跃祖,傅锦业,赵泽明,等.去甲斑蝥素对人胆囊癌GBC-SD细胞系增殖及侵袭的影响[J].中华肿瘤杂志,2004,26(5):271-274.
[10] 范跃祖,赵泽明,陈春球,等.去甲斑蝥素对荷瘤 裸鼠胆囊癌移植瘤的抗癌作用机制探讨[J].中华外科杂志,2006,44(9):618-622.