实时荧光定量PCR检测野生型p53介导蛋白磷酸酶mRNA在非小细胞肺癌中的表达

Expression of Wip1 mRNA in Non-small Cell Lung Cancer by Real-time PCR

【目的】荧光染料Ⅰ(SYBR GreenⅠ)是一种较为常用的非探针类定量PCR的方法,其结果具有一定特异性。本研究的目的是建立检测野生型p53介导蛋白磷酸酶(Wip1)mRNA的SYBR GreenⅠ实时定量PCR的方法,了解肺癌组织及其远癌正常肺组织中Wip1的表达水平,研究肺癌组织中Wip1的表达水平与各种临床病理特征之间的关系。【方法】利用相对定量的方法检测44例肺癌及相应远癌正常肺组织中Wip1mRNA相对于β-actin mRNA的表达量,进行相对定量分析。【结果】44例非小细胞肺癌(NSCLC)及相应远癌正常肺组织均有Wip1mRNA的表达,其中17例非小细胞肺癌中Wip1mRNA高表达,占38.6%,两者表达差异有统计学意义(Ratio=2.1644±1.394,P<0.05);分化程度低的肺癌组织中Wip1mRNA表达量显著高于分化程度高者(F=5.08,P=0.013)。【结论】SYBR GreenⅠ实时定量PCR方法可以成功地检测Wip1基因的表达量。初步检测结果显示Wip1可能是一种肺癌发生发展过程中的致癌因素。

[Objective] The aim of this study was to establish a quantitative SYBR Green Ⅰ real-time PCR method for detection of wide-type p53-induced phosphatase 1 （Wipl or PPM1D） gene expression level in non-small cell lung cancer （NSCLC）, and to investigate the relationship between Wipl mRNA expression level and the clinicopathological characters. [Method] Real-time PCR was employed to determine the expression level of Wipl mRNA in 44 specimens of NSCLC tissues and their adjacent normal tissues. [Results] In the 44 specimens, the expression of Wipl mRNA in both cancer tissues and adjacent normal lung tissues were positive. Wipl gene was overexpressed in 17 specimens among 44 NSCLC specimens. The rate was 38.6%. The relative level of Wipl mRNA in NSCLC tissues was significantly higher than the adjacent normal lung tissues （Ratio = 2.1644 ± 1.394, P 〈 0.01）. The expression of Wipl mRNA was also correlated with pathological staging （F = 5.08, P = 0.013）. [Conclusion] The established SYBR Green Ⅰ quantitative real-time PCR method can successfully detect the expression level of Wipl mRNA. The results suggested that Wipl may be involved in the development of NSCLC.