摘要
目的构建galectin-9基因不同亚型的真核表达载体，观察galectin-9对结肠癌LoVo细胞体外黏附侵袭能力的影响。方法构建galectin-9基因不同亚型的真核表达载体后，用脂质体法转染结肠癌LoVo细胞，逆转录一聚合酶链反应（RT-PCR）和Western blot方法验证转染效果。LoVo细胞分别在转染galectin-9、转染galectin-9＋抗体（5mg／L）和转染galectin-9＋β-乳糖（30mmol／L）条件下进行体外细胞-基质黏附实验和体外细胞侵袭实验。结果测序结果证实载体上插入的galectin-9三种剪接体碱基序列正确，同时载体成功转染LoVo细胞。体外细胞-基质黏附实验中，转染空载体，转染galectin-9L，galectin-9M和galectin-9S组平均吸光度分别为：0．74±0．04、1．14±0．12、1．12±0．10和1．15±0．13，表明galectin-9促进LoVo细胞黏附于细胞外基质（P〈0．05）。galectin-9抗体和β-乳糖可抑制该作用，但转染galectin-9L和galectin-9S LoVo细胞的黏附作用受β-乳糖抑制更明显（P〈0．05）。体外侵袭实验未发现galectin-9对LoVo细胞侵袭力有影响。结论ga-lectin-9促进LoVo细胞与细胞外基质的黏附，这可能与结肠癌侵袭转移相关，但其三种亚型发挥作用的机制不全相同。
Objective To construct three isoforms of galectin-9 gene expression vectors and study the effects of galectin-9 isoformson adhesion and invasion of colon carcinoma LoVo cell hne in vitro. Methods After three isoforms of galectin-9 expression vectors were constructed, the vectors were transfected into LoVo cells by Lipofectamine 2000, and the effect of transfection was confirmed by RT-PCR and Western blot analysis. Cell-matrix adhesion assay and cell invasion assay were performed. In addition, the influence of anti-galectin-9 antibody （5 mg/L） and β-lactose （30 mmol/L） was also investigated. Results Base sequences of three isoforms of galectin-9 were correct and the expression vectors were transfected into LoVo cells successfully. In cell-matrix adhesion assay, the average A value of LoVo-vector, LoVo-galectin-9L, Lo- Vo-galectin-9M and LoVo-galectin-9S was 0.74 ±0.04,1.14 ± 0.12,1.12 ± 0.10 and 1.15 ±0.13,respectively. The results showed all isoforms of galectin-9 facilitated LoVo cells adhere to extracellular matrix （P 〈 0.05 ）, both anti-galectin-9 antibody and β-lactose inhibited the effect compared with control trans- fectants. However, the adhesion of LoVo cells transfected with galectin-9L or galectin-9S was impaired by β-lactose more significantly （ P 〈 0.05 ）. Cell invasion assay revealed that galectin-9 was not correlated to the invasiveness of LoVo cells. Conclusion Ectopic expression of galectin-9 in LoVo cells facihtates the cell-matrix adhesion,which may be involved in tumor metastasis,but the mechanisms through which three isoforms of galectin-9 modulate cell-matrix adhesion may not be the same.
出处
《中华实验外科杂志》
CAS
CSCD
北大核心
2008年第3期288-290,共3页
Chinese Journal of Experimental Surgery
关键词
半乳凝素
结肠肿瘤
黏附
侵袭
Galectin
Colon neoplasm
Adhesion
Invasion
