非小细胞肺癌患者血中DNA片段的检测、定量及临床意义的研究

The study of quantify and detection and clinical significance of plasma free-DNA part of non small cell lung cancer patients

目的探讨非小细胞肺癌患者血中游离DNA片段在临床上早期诊断、分期、预后方面的意义。方法随机采集2004年10月至12月大连医科大学附属二院经病理确诊的初治Ⅲ、Ⅳ期非小细胞肺癌患者的血标本15份,门诊健康体检者血标本15份,通过酚/氯仿提取法定量检测两组的血浆DNA片段,进行对照比较,分析非小细胞肺癌组内各因素(包括与诊断相关的肿瘤标记物,与预后相关的年龄、性别、一般状况及病理类型、疾病分期、免疫指标、是否转移等)与血浆游离DNA片段水平的相关性。结果非小细胞肺癌组血浆游离DNA片段水平高于正常健康对照组,其血浆游离DNA片段的均值分别为:(384.8±99.36)μg/ml与(13.9±3.60)μg/ml,差异有显著性,Ⅲ期与Ⅳ期非小细胞肺癌患者的血浆游离DNA片段差异有显著性,均值分别为:(290.4±91.82)μg/ml与(481.2±196.44)μg/ml,且非小细胞肺癌患者的血浆游离DNA片段与肿瘤标记物CA199、CYFRA21-1、ALP呈正相关性;在预后因素分析中,显示非小细胞肺癌患者的血浆游离DNA片段水平与一般状态、免疫状态、病理类型及是否存在远处转移等因素有明显相关性,其中与一般状态、免疫状态呈负相关,与远处转移呈正相关,与性别、年龄、治疗疗效等因素无相关性。结论非小细胞肺癌患者血中的游离DNA片段水平较健康人高,且不同病期有一定差异,可能与患者自身状态、病理分型、疾病状态有关;游离DNA片段可以作为检测非小细胞肺癌的特异性指标,还可以替代一般状态、免疫状态、病理类型、是否存在远处转移等指标评价远期预后,并有取材方便、检测时间短、可以定量分析的优点,对临床有一定的指导意义和应用价值。

Objectives Through the variation of its srecific circulating DNA,with might be adopted as an efficient marker concering early dianosis ,judgement on the therapeutic effects and recurrence/metastasis follow-up, with actual clinical importance to increase curative/survival rate for cancer patients. Methods The study included 15 cases of Ⅲ, Ⅳ stage of non small cell lung cancer （ NSCLC ） with pathological diagnosis by Dahan Medical University Affiliated 2th Hospital, 2004. 10 - 2004.12, 15 cases of healthy individuals. Quantitative detection of plasma free-DNA part was done in 30 cases by phenol/chloroform collection, and the relationship was analysed between clinical factors of NSCLC ,included cancer maker CEA, CA199, CYFRA21-1, ALP, which were related to ehnieal diagnosis, and age, sex, gain, pathology, stage, immune, metastasis ,effect of therapy, which were related to prediction of disease progression, and the level of plasma free-DNA part. Results It was observed that the level of plasma free-DNA part of NSCLC [ （ 384.8± 99.36 ） μg/ml ] was higher than healthy controls [ （ 13.9 ±3.60） μg/ml], and there was significantly difference. The level of plasma free-DNA part of 11[ stage of NSCLC [ （ 290. 4 ± 91.82） μg/ml] significantly elevated as compared to that of IV stage （481.2 ± 196. 44μg/ml） . There was positive relationship between the level of plasma free-DNA part of NSCLC and the tumor markers CA199, CY- FEA21-1 ,ALP. Moreover, the level of plasma free-DNA part of NSCLC had significant relationship with the general status, immune status, pathology, metastasis, the negative relationship with general status,immune status and the positive with metastasis, no significant relationship with sex and age and therapy. Conclusions The data demonstrated that the level of plasma free-DNA part of NSCLC is higher than that of healthy controls, and the difference is also observed between Ⅲ stage and Ⅳ stage NSCLC. It is possibly related with the general status and pathology and disease progression. Therefore, the plasma free-DNA part is a special marker of the diagnosis and prediction of disease progression of NSCLC. The technology has chnical sense and apphcation for many advantages, for example convenience and rapidity and quantity.