论文部分内容阅读
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)。
GST-π and MDR1 primers were designed by using β-actin as an internal reference, and simultaneous detection of GST -π and MDR1 mRNA expression levels by multiplex quantitative PCR. The optical density of the amplified bands was detected by gel imaging system, and the relative gene expression level was calculated by the optical density ratio of the target gene and β-actin. The method of high sensitivity, good repeatability, can make batch analysis. Indoor X control chart can effectively control quantitative PCR amplification and electrophoresis quality, to ensure that this method can detect less than 2 times the expression difference. The median of GST-π and MDR1 mRNA in breast cancer tissues were 0.27 and 1.32, respectively, and the expression level was positively correlated (r = 0.323, P <0.05).