Isolation of a human gallbladder cancer cell clone with high invasive phenotype in vitro and metastatic potential in orthotopic model and inhibition of its invasiveness by heparanase antisense oligodeoxynucleotides
Isolation of a human gallbladder cancer cell clone with high invasive phenotype in vitro and metastatic potential in orthotopic model and inhibition of its invasiveness by heparanase antisense oligodeoxynucleotides
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DOI：
10.1007/s10585-006-9053-7
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发表时间：
2007-03-01
影响因子：
4
通讯作者：
Shao, Zhi-Min
中科院分区：
文献类型：
--
作者：
Chang, Xin-Zhong;Wang, Zhan-Min;Shao, Zhi-Min
迄今为止，胆囊癌转移的机制仍不清楚。人们对这一问题的了解不足，部分原因是缺乏理想的细胞系和动物模型来进行研究。本研究从人胆囊癌细胞GBC-SD中分离得到21个细胞克隆，并筛选出具有高侵袭表型的细胞克隆GBC-SDHi。 GBC-SDHi 的侵袭表型和转移潜力在新型裸鼠胆囊癌手术原位植入模型中得到证实。乙酰肝素酶是一种降解硫酸乙酰肝素的糖苷内切酶，是肿瘤转移和血管生成的关键介质。 RT-PCR、实时RT-PCR和Western blot结果显示GBC-SDHi细胞中乙酰肝素酶的表达水平与其亲代细胞有显着差异。 GBC-SDHi细胞经反义寡核苷酸处理后，乙酰肝素酶mRNA和蛋白表达显着降低，其体外侵袭能力受到剂量依赖性抑制。该研究为人类胆囊癌的转移研究提供了有用的细胞克隆和临床相关的原位肿瘤模型。还评估了乙酰肝素酶在胆囊癌中的作用。
The mechanisms involved in gallbladder cancer metastasis still remain unclear to date. The poor understanding is due, in part, to the lack of ideal cell line and animal model for study. In the present study, 21 cell clones were isolated from the human gallbladder carcinoma cells GBC-SD and the cell clone GBC-SDHi with high invasive phenotype was fished out. The invasive phenotype and metastatic potential of GBC-SDHi were confirmed in a novel surgical orthotopic implantation model of gallbladder cancer in nude mice. Heparanase, an endoglycosidase that degrades heparan sulfate, is a critical mediator of tumor metastasis and angiogenesis. RT-PCR, real time RT-PCR and western blot showed that the expression levels of heparanase were significant difference between GBC-SDHi and its parent cells. After treated with antisense oligodeoxynucleotides, the heparanase mRNA and protein expression in GBC-SDHi cells were significantly decreased and its invasive potential in vitro was inhibited in a dose-dependent manner. The study provides a useful cell clone and a clinically relevant orthotopic tumor model for the metastatic study in human gallbladder cancer. The roles of heparanase in gallbladder cancer are also evaluated.