不同mRNA差异显示法筛选胃癌细胞耐药相关基因的研究

Screening of multidrug-resistance-related genes in gastric cancer SGC7901 cell line with different mRNA differential display methods

摘要: 目的　比较银染和放射自显影mRNA差异展示方法在分离、克隆胃癌细胞耐药相关基因过程中的优缺点。方法　长春新硷诱导人胃癌SGC790 1细胞, 建立稳定的耐药细胞亚系SGC790 1/VCR。同时应用银染和放射自显影的mRNA差异展示方法筛选胃癌SGC790 1细胞和耐药细胞亚系SGC790 1/VCR之间mRNA表达的差异。结果　显示银染法省时, 花费少, 操作方便 ;在获得差异条带方面放射自显影法明显优于银染法, 且其能得到较多低丰度的差异表达基因 ;而两者在二次PCR扩增, 克隆, 测序, 杂交鉴定及假阳性率方面无明显差异。结论　银染mRNA差异展示法适合于高丰度差异表达基因的快速筛选, 而放射自显影mRNA差异展示则能较彻底地显示两种细胞中多数mRNA的差异

Abstract: Objective The comparison of Staining silver mRNA differential display with autoradiography mRNA differential display method was performed during Isolating and cloning the multidrug-resistance-related genes in gastric cancer SGC7901 cell line. Methods To establish drug-resistance human gastric cancer SGC7901/VCR cell induced by vincristine. Different drug-resistance related genes in SGC7901/VCR were screened with staining silver and autoradiography mRNA differential display method. Results The mRNA di...