Spink8基因对EC9706细胞增殖、凋亡及迁移能力的影响*
Effects of Spink8 gene on proliferation,apoptosis and migration of EC9706 cells

目的:探讨Spink8基因对人食管癌EC9706细胞增殖、凋亡及迁移能力的影响。方法:利用DNA重组技术构建Spink8真核表达重组质粒并转染EC9706细胞,构建稳定表达Spink8的EC9706细胞。取未转染、转染空质粒和稳定表达Spink8的EC9706细胞,采用RT-PCR和Western blot 法检测Spink8 mRNA和蛋白,MTT法检测细胞增殖,Annexin V-APC/7-AAD法检测细胞凋亡,并进行Transwell细胞迁移实验。结果:与未转染和转染空质粒组细胞比较,稳定表达Spink8的EC9706细胞增殖受到抑制(P＜0.05),增殖抑制率为24.5%; 细胞凋亡率增加(P＜0.05); Transwell小室穿膜细胞数减少(P＜0.05)。结论:Spink8可能是一个新的抑癌基因。
Aim: To investigate the effects of Spink8 gene on proliferation, apoptosis and migration of human esophageal cancer cell EC9706.Methods: Eukaryotic expression plasmid pEGFP-Spink8 was established by DNA recombination in vitro, and transfected into EC9706 cells. The expressions of Spink8 mRNA and protein were examined by RT-PCR and Western blot. MTT method was used to detect cell proliferation, Annexin V-APC/7-AAD method was used to detect cell apoptosis, and migration ability was detected by Transwell assay.Results: The plasmid of pEGFP-Spink8 was successfully established. Compared with the EC9706 cells without transfection or transfected with null plasmid, the number of surviving cells transfected with pEGFP-Spink8 decreased(P＜0.05), and proliferation inhibition rate was 24.5%; the apoptosis rate was significantly increased(P＜0.05); the number of cells which passed Transwell chamber decreased(P＜0.05).Conclusion: Spink8 may be a new tumor suppressor gene