摘要
目的研究多药耐药相关蛋白2(MRP2)基因及MRP3、MRP5基因在正常胃细胞系GES-1、胃癌细胞株(BGC-823)及胃癌耐药细胞株BGC-823/ADM的表达差异,并探讨其在胃癌耐药中的意义。方法分别提取GES-1、BGC-823及BGC-823/ADM细胞的总RNA,逆转录cDNA,利用实时荧光定量PCR,根据标准品绘制标准曲线,检测MRP2、MRP3、MRP5基因的表达水平。结果MRP2基因在胃癌耐药细胞株BGC-823/ADM细胞中的表达量明显高于在胃癌细胞株(BGC-823)的表达,而与其在正常胃细胞系GES-1的表达没有显著性差异;MRP3、MRP5基因表达量在三种细胞中均有显著性差异。结论MRP2可能参与了胃癌的内在性耐药,而MRP3、MRP5可能与胃癌的获得性耐药有关,实时荧光定量方法是一种灵敏度高,特异性强,重复性好的定量检测方法。
Objective To investigate the differential expression of genes encoding multidrug resistance-associated protein 2 （MRP2）,muhidrug resistance associated protein 3 （MRP3） and multidrug resistance-associated protein 5 （MRPS） in human normal cell line GES-1 ,gastriccarcinoma cell line BGC-823 and its adriamycin-resistant counterpart BGC-823/ADM cell line. Methods Total RNA was extracted from GES-1 ,BGC-823 and BGC-823/ADM ceils respectively, mRNA was transcribed reversely into cDNA. The expression levels of genes encoding MRP2, MRP3,MRP5 were detected according to the standard curve using real-time fluoregenic quantitative PCR. Results The expression of MRP2 mRNA in the BGC-823/ADM cell line was significantly higher than that in the GES-1 and BGC-823 cell line. There was no significant variation in the expression of MRP2 mRNA between GES-1 and BGC-823 cell line. Whereas, the expression of MRP3 and MRP5 mRNA showed statistically significant difference among 3 cell lines. Conclusion MRP2 might play a role on the intrinsic drug resistanee;MRP3 and MRP5 are related to the acquired drug resistance of gastriccarcinoma.Real-time fluorescent quantitative RT-PCR has good sensitivity, specificity and reproducibility in determining mRNA levels.
出处
《中国现代医药杂志》
2009年第8期8-11,共4页
Modern Medicine Journal of China
关键词
胃癌
多药耐药相关蛋白
实时荧光定量
Gastric carcinoma Muhidrug resistance-associated protein Real-time fluorescent quantitative （RT- PCR）
