不同mRNA差异显示法筛选胃癌细胞耐药相关基因的研究

 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

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

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 differential display method with silver staining was short time, lower cost, and convenience for acting. Autoradiography mRNA differential display method, which can get more differential low|abundance DNA bands, was superior to silver staining method. There was no difference during re|amplifying of differential DNA bands, cloning, sequencing, hybridization for identification and false positive rate using them. Conclusion The silver staining mRNA differential display method was better for rapid screening of high|abundance differential expressed genes; the later was benefited for visualizing more differential expressed genes between SGC7901 cell and SGC7901/VCR cell.