EIF4A1在胃癌组织中表达及其抑制剂对人胃癌细胞株增殖、存活、侵袭、迁移能力影响观察

Expression of EIF4A1 in gastric cancer and effects of EIF4A1 inhibitor on proliferation,invasion,and migration of human gastric cancer cells

目的观察真核翻译起始因子4A1(EIF4A1)在胃癌组织中的表达变化;另观察EIF4A1抑制剂(Silvestrol)对人胃癌细胞株SGC-7901增殖、存活、侵袭、迁移能力影响,并分析其可能机制。方法淋巴结转移阳性、阴性胃癌组织及匹配癌旁正常组织各20例份,采用免疫组化法检测各组织EIF4A1。取对数生长期SGC-7901细胞并分为3组,药物组加含120μg/m L Silvestrol(在DMSO溶剂溶解)的细胞培养液100μL,模型组加含0. 5%DMSO的细胞培养液100μL,对照组加细胞培养液100μL; CCK-8法检测细胞增殖活性,CountessⅡFL细胞计数仪计数活细胞数,Transwell实验检测细胞侵袭能力,划痕实验检测细胞迁移能力,qRT-PCR法和Western blotting法分别检测EIF4A1和AKT mRNA、蛋白。结果与癌旁正常组织比较,淋巴结转移阳性胃癌组织中EIF4A1高表达例数多(P<0. 05)。各组OD值及活细胞数两两比较,P均> 0. 05;与模型组、对照组比较,药物组侵袭细胞数少,细胞迁移距离长,EIF4A1和AKT蛋白、mRNA表达降低(P均<0. 05)。结论淋巴结转移阳性胃癌组织中EIF4A1表达量高于癌旁正常组织; Silvestrol不影响SGC-7901细胞的增殖、存活能力,但可抑制细胞侵袭、迁移能力,其机制可能与Silvestrol调控AKT信号通路有关。

Objective To observe the expression of eukaryotic translation initiation factor 4A1(EIF4A1)in gastric cancer tissues and the effect of EIF4A1 inhibitor(Silvestrol)on the proliferation,survival,invasion and migration abilities in the human gastric cancer cells SGC-7901,and to analyze its possible mechanism.Methods The expression of EIF4A1 was examined by immunohistochemical method in 20 cases of lymph node metastasis positive GC(gastric cancer)tissues and 20 cases of lymph node metastasis negative GC tissues.SGC-7901 cells of logarithmic growth phase were divided into 3 groups:the drug group,the model group,and the control group.The cell culture medium(100μL)containing 120μg/mL Silvestrol(dissolved in DMSO solvent)was added to the cells in the drug group,100μL of cell culture medium containing 0.5%DMSO was added to the model group,and 100μL of cell culture medium was added to the control group.The cell proliferation was evaluated by CCK-8 assay.The alive cells were detected by the CountessⅡFL.The cell invasion ability was evaluated by Transwell assay.The cell migration ability was evaluated by Scratch test.The expression of EIF4A1 and AKT mRNA was examined by qRT-PCR.The protein levels of EIF4A1 and AKT were analyzed by Western blotting.Results Compared with non-malignant adjacent tissues samples,the lymph node metastasis-positive gastric cancer tissues had more cases of high EIF4A1 expression(P<0.05).No significant difference was found in the OD value and number of living cells between every two groups(P<0.05).Compare with the model group and control group,the ability of invasion and migration of cells significantly decreased,and the protein and mRNA expression of EIF4A1 and AKT significantly decreased in the drug group(all P<0.05).Conclusion The lymph node metastasis positive GC tissues had higher EIF4A1 expression than the non-malignant adjacent tissues,and Silvestrol did not affect the proliferation and viability of SGC-7901 cells,but inhibited the invasion and migration of SGC-7901 cells by regulating AKT signaling pathway.