摘要
用β－肌动蛋白（β－ａｃｔｉｎ）作内参照，设计不同扩增片段的谷胱苷肽－Ｓ转移酶（ＧＳＴ－π）和多药耐药基因（ＭＤＲ１）引物，建立同时定量检测ＧＳＴ－π和ＭＤＲ１ｍＲＮＡ表达水平的复合定量ＰＣＲ法。扩增条带光密度值用凝胶成像系统检测，基因相对表达水平用目的基因和β－ａｃｔｉｎ光密度比值计算。本法灵敏度高、重复性好，能作批量分析。室内Ｘ控制图可有效控制定量ＰＣＲ扩增和电泳质量，保证本法能检测出小于２倍的表达差异。在乳腺癌组织中ＧＳＴ－π和ＭＤＲ１ｍＲＮＡ中位数分别为０．２７和１．３２，且表达水平呈明显正相关（ｒ＝０．３２３，Ｐ＜０．０５）。
Based on the principle that the quantity of PCR product is a reflection of the quantity of template at the exponential increasing stage. A reverse transcriptionpolymerase chain reaction(RTPCR) quantitative technique of glutathione Stransferase isoenzyme(GST)and multidrug resistance gene(MDR1)expression was developed through the setting the housekeeper gene,actin and scanning the total optical density of productive bands on the gel.The individual Xcontrol method of complex quantitative PCR was established too.The RTPCR described here is a sensitive,reproducible,nonradioactive method.Using the present technique,71 samples of breast carcinoma were examined.The median of GST and MDR1 was 027 and 132,at the same time,it's expression lever have positive correlation in breast carcinoma.The quantitative RTPCR method can be applied to show the tiny difference of the GST and MDR1 mRNA expression in breast carcinomas.
出处
《临床检验杂志》
CSCD
北大核心
1999年第4期197-199,共3页
Chinese Journal of Clinical Laboratory Science
关键词
聚合酶链反应
乳腺癌
GST-Π
MDR1
quantitative PCR
glutathione Stransferase
*2]multidrug resistance gene
quality control
