摘要
研究磺酰化乳糖基神经酰胺 (SM 3 )的表达水平与肝癌细胞转移潜能的关系 .采用免疫荧光染色和细胞酶联免疫两种方法 ,观察了不同转移潜能的肝癌细胞中磺酰化乳糖基神经酰胺的表达水平 ,发现高转移性肝癌细胞表达SM 3的水平明显高于低转移性肝癌细胞 .高转移性肝癌细胞经过 10 μmol/L维甲酸处理 ,抑制肝癌细胞的转移潜能以后 ,SM 3的表达水平显著下降 (P <0 0 5 ) ,接近低转移性肝癌细胞水平 ,但是SM3的前体分子半乳糖基神经酰胺水平并不降低 ,提示磺酰化作用受到了抑制 .通过脑苷脂磺酰基转移酶 (CST)基因的转染 ,使Hep3B肝癌细胞高表达SM3 .发现CST转染的肝癌细胞与转移相关的行为显著改变 ,经黏附试验发现 ,CST基因转染的肝癌细胞对层粘连蛋白和玻连蛋白 (vitronectin)的黏附显著增强 ,比未转染的或转染载体的肝癌细胞升高 3～ 4倍 .裸鼠体内试验表明 ,这种具有高度表达CST基因的Hep3B细胞比对照组细胞具有更高的转移能力 (P <0 0 5 ) .
The relationship between sulfated lactosyl ceramide expressional level and hepatocellular carcinoma metastasis potential was investigated. By means of immunofluorescent staining and cell ELISA with monoclonal antibodies against sulfated lactosyl ceramide or galactocerebroside, it was observed that the expression of sulfated lactosyl ceramide in high metastasis hepatocarcinoma cells was significantly higher than those in low metastasis hepatocarcinoma. cells. After the hepatocarcinoma cells with high metastatic potential were treated with 10 mumol/L retinoic acid, the level of sulfated lactosyl ceramide on the cell surface was significantly reduced (P< 0.05) to the level closing to the low metastatic hepatocarcinoma cells. The level of galactocerebroside which was the precursor of sulfated lactosyl ceramide did not change significantly. This indicated that the sulfation was inhibited after retinoic acid treatment. Therefore the cerebroside sulfotransferase gene were transferred into hepatocarcinoma cells and selected sulfated lactosyl ceramide-highly-expressed clones. Through adhesive assay, it was observed that the control cells were quite poor in adhesive to vitronectin and laminin, the transfected cells were significantly enhanced in the adhesion to vitronectin and laminin. After 45 minute incubation, the adhesive percentage of the cells was 3 &SIM; 4 time higher in CST-transfected cells than those in control cells. By animal experiments there were more metastatic foci in the liver or lymph nodes in CST-transfected groups than the control (P<0.05). The results suggested that the high expression of SM3 in hepatocarcinoma cells promoted metastasis in nude mice and enhanced the adhesion to vitronectin.
出处
《生物化学与生物物理进展》
SCIE
CAS
CSCD
北大核心
2003年第3期395-400,共6页
Progress In Biochemistry and Biophysics
基金
国家自然科学基金资助项目 ( 30 0 70 183)
上海市教委重点学科基金资助项目 (B980 80 10 )~~
关键词
磺酰化乳糖基神经酰胺
表达水平
肝癌细胞
转移潜能
sulfated lactosyl ceramide
cerebroside sulfotransferase
hepatocarcinoma metastasis
