NHE1反义基因磁性纳米微粒对胃癌细胞NHE1蛋白表达及裸鼠成瘤力的影响

摘要

目的:探讨NHE1反义基因磁性纳米微粒对胃癌细胞NHE1蛋白表达及裸鼠成瘤力的影响,探索胃癌基因治疗新方法.方法:构建NHE1反义基因真核表达载体和NHE1反义基因磁性纳米微粒,将NHE1反义基因转染至SGC-7901胃癌细胞中,通过免疫组织细胞化学SP法检测NHE1蛋白表达情况,流式细胞仪检测细胞凋亡及细胞周期,观察双层软琼脂集落形成能力及转染前后生长曲线的变化.电镜水平比较观察转染与未转染细胞形态学变化,并观察裸鼠成瘤能力的改变.结果:氧化铁磁性纳米颗粒成功地将NHE1反义基因转染至SGC-7901胃癌细胞中,转染NHE1反义基因的SGC-7901胃癌细胞NHE1蛋白表达降低,细胞生长速度减慢、体外生长增殖能力降低,透射电镜下细胞线粒体数量增多、体积增大,内质网扩张,核偏向一侧,极性增强,裸鼠成瘤能力下降.结论:NHE1反义基因磁性纳米微粒能使SGC-7901胃癌细胞的NHE1蛋白表达明显下调,裸鼠成瘤能力降低,NHE1蛋白在胃癌细胞生长中起重要作用,NHE1 基因可能成为基因治疗胃癌的一个理想靶点.%Objective:To study the effects of antisense NHE1 gene magnetic nanoparticles on the expression of NHE1 protein of SGC -7901 gastric cancer cell and the changes of carcinoma forming capacity in nude mouse after transplantationed and its potential role in the genetherapy for gastric cancer. Methods: Antisense NHE1 eukaryotic expression vector was constructed with recombinant DNA technique and transfected into the cells of gastric carcinoma cell line SGC -7901 with liposome DOTAP. The dextran coated iron oxide nanoparticles( DCIONP) was synthesized with deposition, then the expression of NHE1 protein was tested with SP and the changes of carcinoma forming capacity in nude mouse was also observed. We examined the morphological change of transfected and untransfected cells under electron microscopes. Results: The expressions of NHE1 protein of SGC - 7901 cells transfected with antisense NHE1 gene magnetic nanoparticies in vitro were obviously decreased. Under electron microscope, organelles in these cells were abundant, carcinoma forming capacity in nude mouse was decreased. Conclusion: The expression of NHE1 protein was obviously down regulated after the transfection of antisense NHE1 geneagnetic nanoparticles, and carcinoma forming capacity in nude mouse was decreased. Which implies that NHE1 protein plays an important role in gastric cancer cell growth.