实时荧光PCR检测宫颈组织TSLC1基因表达

目的建立检测肺癌肿瘤抑制因子1（TSLC1）基因mRNA表达的实时荧光聚合酶链反应（PCR）方法。方法设计特异性引物和TaqMan探针,以3-磷酸甘油醛脱氢酶（GAPDH）基因作为对照,建立检测TSLC1基因表达的实时荧光PCR方法,并采用细胞株和宫颈组织样本进行验证。结果该方法检测TSLC1基因mRNA的检测下限为每反应10个细胞或2拷贝cDNA,总不精密度为3.1%。TSLC1基因mRNA在子宫肌瘤、低度鳞状上皮内病变（LSIL）、高度鳞状上皮内病变（HSIL）与宫颈癌患者宫颈组织中的表达率分别为100%、75%、55%和24%。宫颈癌细胞株中未检出TSLC1基因mRNA表达。所有样本中均有GAPDH基因mRNA表达。结论该实时荧光PCR方法能够有效检测TSLC1基因mRNA表达,是进一步研究TSLC1基因在宫颈癌发生中的作用及宫颈癌早期诊断的重要工具。

Expression of TSLC1 gene in cervical tissue by real-time fluorescence PCR

Objective To establish real-time fluorescence polymerase chain reaction (PCR) method for the detection of tumor suppressor in lung cancer 1 (TSLC1) mRNA expression. Methods The real-time fluorescence PCR method for the detection of TSLC1 gene expression was established with designed specific primers, TaqMan probe and glyceraldehyde 3-phosphate dehydrogenase (GAPDH) gene as control, and verified by cell lines and cervical tissue samples. Results The detection limit of TSLC1 gene mRNA was 10 cells per reaction,or 2 copies of cDNA. The total imprecision of this method was 3.1%. The TSLC1 gene mRNA expression rates in cervical tissue of the patients with hysteromyoma, low-grade squamous intraepithelial lesion (LSIL), high-grade squamous intraepithelial lesion (HSIL) and cervical cancer were 100%, 75%, 55% and 24%,respectively. TSLC1 gene mRNA expression was not found in cervical cancer cell lines. GAPDH gene mRNA expression was positive in all samples. Conclusions The real-time fluorescence PCR method can effectively detect mRNA expression of TSLC1 gene and is an important tool for a further study of the effect of TSLC1 gene in cervical cancer development and the early diagnosis of cervical cancer.