骨肉瘤外周血循环肿瘤细胞检测的临床意义

Clinical significance of peripheral blood circulating tumor cells detection in osteosarcoma patients

目的探讨骨肉瘤患者外周血循环肿瘤细胞（circulating tumor cell，CTC）计数、分型与患者病情、疗效及预后之间的关系。验证CTC作为判断骨肉瘤转移、疗效评估及预测患者预后的辅助手段的可行性。方法回顾性分析2015年1月至2016年6月病理确诊的新发骨肉瘤患者30例，男17例，女13例；年龄7-21岁，平均（13．77±4．31）岁；Ennecking Ⅱ期21例，其中ⅡA期2例，ⅡB期19例，Ⅲ期9例。所有患者均在活检手术时、新辅助化疗后及肿瘤切除术后分别进行循环肿瘤细胞检测。循环肿瘤细胞富集采取滤膜过滤技术，利用多重RNA原位分析技术对滤膜截留下的细胞进行鉴定与分型。结合临床资料，对比分析骨肉瘤患者外周血CTC计数、计数变化、分子学特征与肿瘤转移、化疗效果、患者预后之间的关系。结果EnneckingⅢ期患者外周血CTC计数为（10．44±5．70）个，Ⅱ期患者为（4．76±3．56）个。CTC计数增加或减少与新辅助化疗效果良好或不良相关（P=0．002），化疗效果良好的患者，化疗后外周血CTC计数减少。术后CTC数量／〉7者平均无进展生存期为（11．85±4．93）个月，CTC数量〈7者平均无进展生存期为（15．53±4．09）个月，差异有统计学意义（P=0．012）。EnneckingⅢ期者体内间质型CTC的比例为40．42％，高于EnneckingⅡB期患者23．00％，差异具有统计学意义（P=0．010）。肿瘤转移相关基因1（Metastasis—associated gene 1，MTA1）在间质型CTC中的阳性表达率为85．50％，其他分型的CTC阳性表达率为66．67％，差异有统计学意义（P=0．000）。结论骨肉瘤患者外周血循环肿瘤细胞的计数、分型可以作为目前常用的影像学检查的补充，在判断肿瘤转移，疗效观察及预测不良预后过程中发挥辅助作用。

Objective To discuss the correlations amongst the count, molecular and clinicopathological characteristics of circulating tumor cells （CTCs）, and the feasibility of using CTC as an aid to judge the metastasis of osteosarcoma, evaluate the ther- apeutic effect, and predict the prognosis of patients. Methods Retrospective analysis of 30 patients with newly diagnosed osteo- sarcoma diagnosed from January 2015 to June 2016, there were 17 males and 13 females. The average age was 13.77＋4.31 years old （ranged from 7 to 21 years）. There were 21 cases of Ennecking stage II, including 2 cases of IIA, 19 cases of IIB, and 9 cases of Ennecking III. Peripheral blood of all patients was obtained and processed using CanPatrolTM System at baseline, pre- and post-op- eration. Multiplex RNA-in situ hybridization （RNA-ISH） assay was used to characterize the molecular markers. Combined with clinical data, the relationship between peripheral blood CTC counts, count changes, molecular characteristics, tumor metastasis, chemotherapy effects, and prognosis of patients with osteosarcoma was analyzed and compared. Results The CTC count in pe- ripheral blood was 10.44±5.70 in Ennecking stage III patients and 4.76±3.56 in stage II patients. The increase or decrease in CTC counts was associated with good or poor neoadjuvant chemotherapy （P=0.002）. In patients with good chemotherapy effect, the CTC count in peripheral blood was decreased after chemotherapy. The mean progression-free survival time was 11.85±4.93 months for patients with CTC number ≥7, and the average progression-free survival time was 15.53±4.09 months for patients with CTC number 〈 7 （P=0.012）. The proportion of interstitial CTC in Ennecking stage III was 40.42%, which was higher than that of Enneck- ing liB stage 23.00%. The difference was statistically significant （P=0.010）. The positive rate of metastasis-associated gene 1 （MTA1） in interstitial CTC was 85.50%, and the positive rate of other types of CTC was 66.67%. The difference was statistically significant （P=0.000）. Conclusion The counting and typing of peripheral blood CTC in osteosarcoma patients can be used as a complement to the commonly used imaging examinations, and they can play an auxiliary role in judging tumor metastasis, curative effect observation and prediction of poor prognosis.