摘要
背景与目的SYBRGreenⅠ是一种较为常用的非探针类定量PCR的方法,其主要优点在于操作过程简单,结果具有一定特异性。本研究的目的是建立检测色素上皮衍生因子(pigmentepitheliumderivedfactor,PEDF)mRNA的SYBRGreenⅠ实时定量PCR的方法,了解肺癌组织及其远癌肺组织中PEDF的表达水平,研究肺癌组织中PEDF的表达水平与各种临床病理特征之间的关系。方法利用RTPCR的方法扩增并纯化PEDF以及内对照GAPDHmRNA的目的片段,倍比稀释后作为定量模板。利用相对定量的方法,检测21例肺癌及相应远癌肺组织中PEDFmRNA相对于GAPDHmRNA的表达量,进行相对定量分析。结果21例肺癌组织及相应的远癌肺组织中均可检出PEDF的表达,肺癌组织中PEDF的相对表达量为0.5505±0.3590(0.11～1.11),远癌肺组织中为0.7219±0.2582(0.29～1.31)(P=0.024)。早期(Ⅰ～Ⅱ期)肺癌患者和肿瘤较小时(T1)PEDF的表达量显著高于晚期(Ⅲ～Ⅳ期,P=0.010)和肿瘤较大者(T2～4,P=0.007)。结论所建立的SYBRGreenⅠ实时定量PCR方法可以成功地检测PEDF基因的表达量。初步检测结果显示PEDF可能是一种肺癌发生发展过程中的保护因素。
Background and objective SYBR GreenⅠ is a non-probe real-time PCR method. It is quite convenient and its specificity is good. The aim of this study is to estahlish a quantitative SYBR GreenⅠ realtime PCR method for detection of PEDF gene in non-small cell lung cancer （NSCLC）, and to investigate the relationship between PEDF mRNA expression and the clinicopathological characteristics. Methods The target segments of PEDF and GAPDH proliferated by RT-PCR were diluted and used as the standard templates. PEDF mRNA was detected in 21 NSCLC tissues and matched paracancerous pulmonary tissues by relative quantitative method. Results All the lung cancer tissues and the matched paracancerous pulmonary tissues had expression of PEDF mRNA. The relative level of PEDF mRNA expression was 0. 5505±0. 3590 （0. 11- 1.11） in the lung cancer tissues and 0. 7219±0. 2582 （0. 29-1.31） in the matched paracancerous pulmonary tissues respectively （P=0.024）. PEDF expression was significantly related to TNM stages （Ⅰ-Ⅱ vs Ⅲ-Ⅳ, P=0. 010） and the tumor size （T1 vs T2-4, P=0.007）. Conclusion The established SYBR GreenⅠ quantitative real-time PCR method can successfully detect the expression of PEDF mRNA. The primary results show that PEDF may be a protective factor in oncogenesis and development of NSCLC.
出处
《中国肺癌杂志》
CAS
2006年第2期177-181,共5页
Chinese Journal of Lung Cancer
基金
英国癌症研究基金(C7603/A3065)资助~~
