CK19实时荧光定量PCR法动态检测非小细胞肺癌患者围手术期外周循环癌细胞  

Detection of circulating tumor cells in patients with non-small-cell lung carcinoma by real time fluorescent quantitative RT-PCR with cytokeratin 19 in perioperative period

作　　者：葛明建[1] 时德[2] 吴庆琛[1] 王梅[3] 李强[1] 李良彬[1] 

机构地区：[1]重庆医科大学附属第一医院胸外科,重庆400016 [2]重庆医科大学附属第一医院普外科,重庆400016 [3]重庆医科大学附属第一医院检验科,重庆400016

出　　处：《肿瘤》2006年第10期914-919,共6页Tumor

基　　金：重庆市卫生局科研基金资助项目(编号:00-2004)

摘　　要：目的:动态观察围手术期非小细胞肺癌(NSCLC)患者外周循环癌细胞(circulating tumor cells,CTCs)的存在状况,评价手术操作促进癌细胞入血的可能性,寻找术中血管结扎顺序与CTCs数量变化问的相关性。方法:按预定的结扎血管顺序术前将23例接受根治性手术的原发性NSCLC患者随机分为先结扎肺静脉组和先结扎肺动脉组,分别采集其术前1天、术中及术后第7天的外周血标本。以角蛋白19(CK19)作为标志物,运用实时荧光定量逆转录-聚合酶链反应(fqRT-PCR)法定量检测各时点外周血中CK19 mRNA水平。选择10例需手术治疗的肺部良性疾病患者作为手术对照;以20例健康人作为阴性对照。结果:肺癌患者CK19术中测值显著高于术前1 d(5．2464±0．196vs 4．472±0．164,P<0．001)及术后7 d(5.246±0．196 vs 4．694±0．177,P=0．013)。肺腺癌与鳞癌比较有显著差异(4．911±1．032 vs 4．1891±0．413,t=2．364,P=0．028)。围手术期先结扎肺静脉组与先结扎肺动脉组比较有明显差异(4．503vs 5．085,P=0．086)。以健康对照组均值的95％可信区间的上界限为分界值,若测得值>4．120为阳性。14例肺癌患者术前结果为阳性(60．9％):2例接受手术的肺部良性疾病患者出现术中正常上皮细胞入血。结论:人表达CK19 mRNA的癌或上皮细胞主要在术中入血。外科手术操作可促使癌细胞术中入血,若先结扎肺静脉可一定程度上减少肺癌细胞入血。Objective： To observe the circulating tumor cells （CTCs） in patients with non small cell lung carcinoma （NSCLC） in perioperative period dynamically, assess whether surgical manipulation could accelerate tumor cells into bloodstream, and determine the association between the sequence of vessel ligation and the quantities of CTCs in circulation. Methods： According to the order of vessel ligation, 23 NSCLC patients undergoing surgical resection with curative intention were randomly assigned to pulmonary vein （PV）-first group and pulmonary artery （PA）-first group before the operation. Peripheral blood samples were collected on d 1 preoperation, intraoperation, and d 7 postoperation. Using cytokeratin 19（CK19）as a marker, CK19 mRNA expression was measured by real time fluorescent quantitative reverse transcriptase-polymerase chain reaction （fqRT- PCR）. Ten patients with benign lung disease who need surgical therapy served as surgery control. Twenty healthy people served as negative control. Results：The level of CK19 mRNA in operation was significantly higher than that on d 1 preoperation （5. 246±0. 196 vs 4. 472±0. 164, P〈0. 001） and on d 7 postoperation （5. 246±0. 196 vs 4. 694±0. 177, P=0. 013）. The difference between adenocar cinoma and squamous carcinoma was significantly different （4. 911±1. 032 vs 4. 189±0. 413, t=2. 364, P=0. 028）. There was signifi cant difference between PV-first group and PA-first group in perioperative period （4. 503 vs 5. 085, P=0. 086）. Using the upper limit of 95% CL of the mean value of healthy control group as threshold, if the determined value was 〉 4. 120, the result was considered positive. Before operation, 14 NSCLS patients （60.9%） were CK19-positive. Surprisingly, circulating epithelial cells is detected in 2 patients with benign lung disease during surgery. Conclusion： The CK19-expressing tumor or epithelial cells were released intraoperatively into the circulation. Surgical manipulation could promote the release of t

关 键 词：肺肿瘤 癌 非小细胞肺 血液 聚合酶链反应 细胞角蛋白 RNA 信使 围手术期 

分 类 号：R734.2[医药卫生—肿瘤] R73-37[医药卫生—临床医学]