支气管肺泡灌洗液脱落细胞端粒酶hTERT基因表达的检测

背景与目的:肺癌患者中端粒酶活性常呈阳性,常规检测端粒酶活性的方法易发生假阳性,端粒酶hTERT基因表达水平可反映端粒酶活性高低。本研究采用相对定量PT-PCR法检测肺癌支气管肺泡灌洗液脱落细胞端粒酶hTERT的表达情况,为肺癌的早期诊断、鉴别诊断提供帮助。方法:采用纤维支气管镜收集了20例可疑肺肿瘤患者支气管肺泡灌洗液,对脱落细胞分别送病理检测和RT-PCR相对定量法检测端粒酶hTERT基因表达,并随访术后病理检测情况,同时检测10例非肺癌病人的支气管肺泡灌洗液标本作为对照。结果:20例可疑肺癌经术后病理检测证实有19例为不同类型的肺癌,1例为炎性假瘤。在19例肺癌的支气管肺泡灌洗液中16例检出不同程度的端粒酶hTERT基因表达,细胞病理检测仅发现11例,阳性率分别为84.2%和57.9%,两者有显著性差异(P<0.01)。10例非肺癌病人的支气管肺泡灌洗液中均无端粒酶hTERT基因表达。结论:采用RT-PCR方法检测支气管肺泡灌洗液中端粒酶hTERT基因表达,较细胞病理检测有更高的敏感性,有助于肺癌的早期诊断和鉴别诊断。

Detection of telomerase hTERT gene expression of exfoliated cell in broncho-alveolar lavage fluid

Background &Objectives:The patients with lung carcinoma usu ally have high telomerase activity,the pseupositive result is happen frequently when the test is done with tradi tional method.Telomerase activity is more strongly correlated with hTERT mRNA.This stu dy was designed to detect the expression of telomerase hTERT gene in broncho-alveolar lavage fluid of th e patients with lung carcinomas by re lative quantitative RT-PCR method,to afford a useful tool for early and differentiate dia gnosis of lung cancer.Method:The exfoliated cells in broncho-alv eolar lavage fluid from20human suspicion lung carcinomas a nd 10health persons were analyzed fo r hTERT expression by relative quant itative RT-PCR method,in the same time a part of exfoliated cells were detected wi th pathologic method.After operati on,pathologic detection was performed for all samp les.Results:Of 20human suspicion lung carcinoma sand,19patients were lung cancer,1patient was inflammatory p seudotumor in pathological diagnosis after operation.The relative exp ression of telomerase hTERT gene in 16cases of 19patients with lung carcinoma were d etected with the relative quantitative RT-PCR,the positive rate is 84.2%(16/19),the expressions of telomerase hTERT gene were at variou s quantities,average levels is 0.42.The positive rate in exfolia ted cells detected with pathologic method were 57.9%(11/19).There was a statistical difference between them(P<0.01).Ten normal persons were 534hTERT gene expression negative.Conclusions:Relative quantitative RT-PCR were more sensitive than cell morphologic method in detecting telomerase hTER T gene expression in broncho-alveolar lavage fluid of patients with lung carcinomas.Detecting quantitatively the levels of telomerase hTERT gene expressio n in broncho-alveolar lavage fluid w ill may be useful for early and differentiate diagnosis of lung carcinomas,especially fo r peripheral lung carcinomas.