乙酰肝素酶基因过表达对小鼠肝癌细胞血道及淋巴道转移的影响

Effects of heparanase over-expression on the hematogenous and lymphatic metastasis of hepatocellular carcinoma cells in mice

目的利用小鼠乙酰肝素酶pcDNA3.1(+)表达载体转染高转移潜能肝癌细胞株(Hca-F)和低转移潜能肝癌细胞株(Hca-P),研究其对小鼠肝癌细胞血道及淋巴道转移的影响。方法构建pcDNA3.1(+)-hpa质粒,以lipofectamine2000作为转染试剂,将其分别转至Hca-F、Hca-P细胞株。转染空载体的肝癌细胞作为对照。血道转移实验:小鼠尾静脉注射肿瘤细胞,30d后观察小鼠的肺部转移瘤生长情况。淋巴道转移实验:在小鼠右下肢爪垫内侧皮下注射肿瘤细胞,30d后观察荷瘤鼠腋窝淋巴结或/和腹股沟淋巴结转移情况。结果尾静脉注射转染有pcDNA3.1(+)-hpa质粒的高转移Hca-F和低转移Hca-P株,荷瘤小鼠的肺表面转移结节数分别为(37.5±2.63)个和(32.6±2.91)个,显著高于其各自空白对照组的(31.3±2.79)个和(14.2±2.38)个(P<0.05)。皮下注射转染有pcDNA3.1(+)-hpa质粒的高转移Hca-F和低转移Hca-P株,荷瘤小鼠淋巴结转移率分别为93.3%和83.3%,显著高于各自空白对照组(P<0.05)。结论乙酰肝素酶基因过表达对小鼠肝癌细胞血道及淋巴道转移均有一定的促进作用。

Objective To transfect hepatocellular carcinoma cell lines of high (Hca-F) and low (Hca-P) metastatic ability with pcDNA3.1(+) expressing heparanase,and investigate its effects on the hematogenous and lymphatic metastasis of hepatocellular carcinoma cells in mice.Methods Mouse hepatocellular carcinoma cell lines Hca-F and Hca-P were transfected with pcDNA3.1(+) expressing heparanase by lipofectamine 2000,with empty pcDNA3.1(+) vector as control.In hematogenous metastasis experiment,the number of the lung superfacial metastases was recorded 30 days after intravenous injection of transfected tumor cells into the mouse tail veins.In lymphatic metastasis experiment,the metastasis of axillary lymph nodes and/or inguinal lymph nodes was observed 30 days after subcutaneous injection of transfected tumor cells to the pad of right lower limb.Results The numbers of the lung superfacial metastases were 37.5±2.63 and 32.6±2.91 in mice intravenously injected with transfected hepatocellular carcinoma cell lines of high(Hca-F)and low(Hca-P) metastatic ability,significantly higher than 31.3±2.79 and 14.2±2.38 in their negative transfected groups respectively(P<0.05).The lymph node metastasis rates were 93.3% and 83.3% in mice subcutaneously injected with transfected Hca-F and Hca-P lines,significantly higher than those in their negative control groups respectively(P<0.05).Conclusion Over-expression of heparanase can promote haematogenous and lymphatic metastasis of hepatocellular carcinoma cells,which may be used as a new therapeutic target.