晚期胰腺癌患者门静脉血循环肿瘤细胞检测的临床应用价值

The clinical value of circulating tumor cells acquired from portal vein of patients with advanced pancreatic cancer

目的探讨介入微创方法获取晚期胰腺癌患者门静脉循环肿瘤细胞(circulating tumor cells,CTCs)的可行性、安全性及其临床应用价值。方法从2016年2月—12月复旦大学附属中山医院介入治疗科收治的晚期胰腺癌患者中纳入29例。所有患者接受动脉化疗灌注或动脉化疗栓塞术前,于超声导引下用21G穿刺针穿刺肝内门静脉,抽取门静脉血15 mL,部分患者同时抽取外周静脉血7.5 mL。ClearCell系统富集CTCs后,EpCAM/CK19/DAPI免疫荧光染色鉴定并计数。比较单位门静脉血样和外周静脉的CTCs计数,分析门静脉CTCs计数和患者临床/病理特征的相关性,观察门静脉和外周静脉CTCs的干细胞表征。结果 29例晚期胰腺癌Z患者均成功获取门静脉血,未出现并发症。29例患者的门静脉CTCs平均计数为(282±50)/7.5 mL。14例同时采集门静脉血和外周静脉血的患者平均门静脉CTCs计数明显高于外周静脉[(237±78)/7.5 mLvs.(21±7)/7.5 mL,P=0.010 2]。伴肝转移患者门静脉CTCs平均计数明显高于无肝转移患者[(449±81)/7.5mLvs.(126±18)/7.5 mL,P=0.000 4]。门静脉CTCs中CD133^+细胞比例为5.9%,外周静脉CTCs中未检测到CD133^+细胞。结论超声导引下21G穿刺针穿刺肝内门静脉,是安全可行的获取晚期胰腺癌患者门静脉血的方法。晚期胰腺癌患者的单位门静脉血CTCs计数明显高于外周静脉,门静脉CTCs计数与是否伴有肝转移相关。晚期胰腺癌患者门静脉CTCs中部分具有干细胞表征。

Objective To investigate the [easibility, safety, and clinical application values of interventional radiology techniques in acquiring circulating tumor cells （CTCs） from portal vein in patients with advanced pancreatic cancer. Methods From Feb., 2016 to Dec., 2016, patients diagnosed with advanced pancreatic cancer in the Department of Interventional Radiology, Zhongshan Hospital,Fudan University were enrolled in this study. Percutaneous transhepatic portal vein puncture were performed with a 21 G needle under the guidance of ultrasound and 15 mL portal vein blood were obtained for each patient before transarterial chemotherapy/chemoembolization. In some patients, 7.5 mL peripheral vein blood was also obtained. CTCs were enriched by ClearCell system. EpCAM/ CK19/DAPI triple positive cells were considered as CTCs and were counted using immunofluorescence assay. CTCs cell counts were compared between the portal vein blood and peripheral vein. The correlations of CTCs and clinical/pathological characteristics of the patients were analyzed. The stem cell characteristic of CTCs was observed both in portal vein and peripheral vein. Results Percutaneous transhepatic portal vein puncture and portal vein blood sampling was successfully performed in 29 pancreatic cancer patients without technique related complications. The mean counts of portal vein CTCs was （282 ＋ 50）/7.5 mL in all 29 patients. In 14 patients with both portal vein and peripheral vein sample obtained,the mean counts of CTCs in portal vein was significantly higher than thatin theperipheralveinblood[（237± 78）/7.5 mL vs. （21±7）/7.5 mL,P=0.010 2）].In portal vein samples,the mean counts of CTCs in patients with liver metastasis was significantly higher than patients without liver metastasis [（449 ± 81）/7.5 mL） vs. （26 ± 18）/7. 5 mL,P=0. 000 4）1. The percetage of CD133^＋ CTCs was 5.90/00 in portal vein,while no CD133^＋ CTCs was found in peripheral vein. Conclusions Percutaneous transhepatic portal vein puncture using a 21 G needle for portal vein blood sampling under the guidance of ultrasound was safe and feasible for patients with advanced pancreatic cancer. CTCs counting was significantly higher in the portal vein than peripheral vein. Portal vein CTCs counting was correlated with liver metastasis status. Stem characteristic CTCs can be found in portal vein.