实时荧光定量端粒重复扩增法检测大肠癌患者单个核细胞端粒酶基因表达

目的 研究大肠癌患者外周血单个核细胞(PBMC)端粒酶基因表达与临床病理的关系.方法 用实时荧光定量端粒重复扩增法(RTQ-TRAP)检测71例大肠癌和20例良性大肠疾病患者,以及25名健康人PBMC端粒酶基因表达,同时检测大肠癌患者血浆癌胚抗原(CEA)含量.结果 71例大肠癌患者中,50例端粒酶基因表达阳性,阳性率70.4%;20例良性大肠疾病表达阳性1例,阳性率为5.0%,大肠癌组与良性大肠疾病组和健康对照组的PBMC端粒酶基因表达差异有统计学意义(χ2=24.521,P<0.001).大肠癌患者在不同性别(χ2=0.114,P=0.736)、年龄(χ2=0.113,P=0.735)、肿瘤部位(χ2=1.523,P=0.677)、Dukes分期(χ2=2.282,P=0.320)间的端粒酶基因表达差异无统计学意义.71例大肠癌患者PBMC端粒酶基因表达阳性率与CEA阳性率(63.4%)比较,差异无统计学意义(χ2=2.286,P=0.125).结论 RTQ-TRAP是一种快速、准确、定量检测端粒酶基因表达的方法,大肠癌患者PBMC端粒酶活性是一项有效的辅助诊断大肠癌的分子标志.

Detection of telomerase gene expression In the peripheral blood mononuclear cells in patients with colorectal carcinoma by real-time quantitative telomeric-repeat amplification protocol assay

Objective To investigates the gene expression of telomerase by real-time quantitative telomeric-repeat amplification protocol assay (RTQ-TRAP) in the peripheral blood mononuclear cells (PBMC) of colorectal carcinoma patients and the relationship between the telomerase gene expression in PBMC and clinicopathological features. Methods Peripheral blood samptes were collected from 71 colorectal carcinoma patients, 20 benign colorectal disease patients and 25 normal controls. The telomerase gene expression in PBMC was measured by RTQ-TRAP, and serum CEA in colorectal carcinoma patients was measured by chemiluminescence immunoassay. Results The gene expression of telomerase in PBMC was positive in 50 out of 71 cancer patients (70. 4% ) , 1 out of benign patients (5. 0% ), respectively. The difference of the telomerase gene expression of PBMC in benign colorectal diseases and cancer patients was significant (χ2 = 24. 521, P < 0. 001 ). There was no significant association between the expression of telomerase and patient's gender, age, Dukes stage, and tumor site. The positive rate of CEA in colorectal carcinoma patients was not significantly higher than positive rate of telomerase gene expression(χ2 = 2. 286,P = 0. 125). Conclusions The RTQ-TRAP method is highly accurate and sensitive in measuring telomerase gene expression. The detection of telomerase gene expression in PBMC of colorectal carcinoma patients is a simple and useful molecular marker for the diagnosis of colorectal carcinoma.