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。
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.
作者
冷娇
刘雪梅
张匠
毛英
刘黎
Jiao Leng;Xue-mei Liu;Jiang Zhang;Ying Mao;Li Liu(Department of Oncology,Suining Central Hospital,Suining,Sichuan 629000,China)
出处
《中国现代医学杂志》
CAS
2020年第8期13-19,共7页
China Journal of Modern Medicine
