Galectin-1 与胃癌细胞阿帕替尼敏感性的 关系及调控机制研究

Relationship between Galectin-1 and sensitivity of gastric cancer cells to apatinib and its possible mechanism

摘要:

目的 探讨半乳糖凝集素1（Galectin-1）与胃癌细胞株SGC-7901 对阿帕替尼敏感性的关 系，以及可能的调控机制。方法 转染Gal1-siRNA 至SGC-7901 细胞，并采用实时荧光定量聚合酶链反应 （qRT-PCR）和Western blotting 验证干扰效果。另外设置空白对照组和阴性siRNA 组。采用MTT 法检测 细胞对阿帕替尼的敏感性；Annexin V/PI 双染法和DAPI 法检测阿帕替尼对细胞凋亡活力的影响；Western blotting 检测各组细胞中上皮间质转化相关蛋白E-cadherin、Vimentin，以及相关转录因子Snail、Twist、ZEB 的表达。结果 不同浓度阿帕替尼作用48 h，或25μmol/L 阿帕替尼分别作用不同时间，Gal1-siRNA1 组细 胞增殖抑制率均高于空白对照组（P <0.05）。25μmol/L 阿帕替尼处理48 h 后，Gal1-siRNA1 组细胞凋亡率 高于空白对照组（P <0.05）。同时与空白对照组和阴性siRNA 组比较，Gal1-siRNA1 组细胞E-cadherin 蛋白 表达升高，而Vimentin 和Snail 蛋白表达降低（P <0.05）。结论 通过siRNA 干扰SGC7901 细胞Galectin-1 mRNA 和蛋白的表达后，细胞对阿帕替尼的敏感性增强，其作用机制可能与抑制上皮间质转化进程有关。

Abstract:

Objective To discuss the relationship between Galectin-1 and sensitivity of gastric cancer cells SGC7901 to apatinib and its possible mechanism. Methods SGC-7901 cells were cultured in vitro. Chemically synthesized siRNA targeting Galectin-1 were transfected into SGC-7901 cells, which were verified by quantitative real time polymerase chain reaction (qRT-PCR) and Western blotting. SGC-7901 cells with the non-transfected cells and cells transfected with negative siRNAs were made as blank group and negative siRNA group. The proliferation of SGC-7901 cells was detected by MTT assay. The apoptosis of SGC-7901 cells was detected by Annexin V/PI staining and DAPI staining. The levels of epithelial cadherin (E-cadherin), Vimentin, Twsit, Snail and zinc finger E-box binding homeobox (ZEB) proteins in SGC-7901 cells were detected by Western blotting. Results After apatinib with different concentrations treated for 48h or 25μmol/L apatinib treated for different time, the inhibition rates of SGC-7901 cells in Gal1-siRNA1 group were higher than that in blank group (P < 0.05). The apoptosis rate of apatinib for 48h on SGC-7901 cells in Gal1-siRNA1 group were higher than that in blank group (P < 0.05). Compared with blank group and negative siRNA group, E-cadherin protein of SGC-7901 cells in Gal1-siRNA1 group was higher,while Vimentin protein and Snail protein were lower (P < 0.05). Conclusions Silencing Galectin-1 can enhance the sensitivity of SGC7901 cells to apatinib, which may be related to inhibiting EMT process via regulating E-cadherin, Vimentin and Snail.