肺癌胸腔积液肿瘤细胞及CD133^+细胞数对病情评估和疗效预测的价值

Clinical significance of tumor cells and CD133 positive cells detection in disease evaluation and curative effect prediction from pleural effusion of patients with lung cancer

目的肺癌胸腔积液的成分复杂,常规方法难以获得高纯度的胸腔积液肿瘤细胞。在前期建立免疫磁珠阴性富集法获得高纯度肿瘤细胞的基础上,本研究进一步探讨恶性胸腔积液中肿瘤细胞及CD133^+细胞数对肺癌患者病情评估及疗效预测的作用。方法收集2013-04-01-2014-08-31扬州大学临床医学院活检病理确诊且伴有恶性胸腔积液的初治肺癌患者21例。治疗前收集10mL胸腔积液,采用阴性富集技术、染色体荧光原位杂交整合技术及CD133抗体免疫荧光染色对肿瘤细胞进行富集和荧光染色并计数。分析其与临床病理特征及治疗疗效之间的关系。结果 21例患者胸腔积液的中位肿瘤细胞数及CD133^+细胞数分别为4 000个/10mL和19个/10mL。肿瘤细胞数及CD133^+细胞数在不同性别(χ~2=0.597,P=0.550;χ2=0.225,P=0.822)、年龄(χ~2=0.533,P=0.594;χ~2=1.288,P=0.198)和病理类型(χ~2=3.253,P=0.197;χ~2=0.034,P=0.983)之间差异无统计学意义。与不伴有或仅伴有区域淋巴结转移患者比较,当肿瘤伴有肺内或远处淋巴结或其他脏器转移时,其胸腔积液中肿瘤细胞数(χ~2=4.398,P=0.036)和CD133^+细胞数(χ~2=4.605,P=0.032)明显增高。全组患者在化疗2个周期或口服吉非替尼2个月后,CR 0例,PR 7例,SD 9例,PD 5例,总有效率为33.33%。不同疗效患者的肿瘤细胞数及CD133^+细胞数差异均有统计学意义(χ2=7.575,P=0.023;χ2=7.247,P=0.027),疗效越好的患者肿瘤细胞数及CD133^+细胞数越少。以肿瘤细胞数及CD133^+细胞数的中位值为界值并联合进行分组,将所有患者分为3组:少量细胞数组(肿瘤细胞数和CD133^+细胞数均≤中位值)、中等细胞数组(肿瘤细胞数>中位值并且CD133^+细胞数≤中位值,或者CD133^+细胞数>中位值并且肿瘤细胞数≤中位值)和大量细胞数组(肿瘤细胞数和CD133^+细胞数均>中位值)。中等-大量细胞数组患者肿瘤伴有肺内或远处淋巴结或其他脏器转移的发生率明显高于少量细胞数组,χ~2=12.634,P=0.002;少量细胞数组患者治疗疗效显著优于中等-大量细胞数组患者,χ~2=10.886,P=0.01。结论胸腔积液中肿瘤细胞和CD133^+细胞数有可能是晚期肺癌患者病情评估和疗效预测有效预测因子。

OBJECTIVE The composition of malignant pleural effusion in lung cancer is so complicated that it is very difficult to get high purity of tumor cells from pleural effusion. Based on the establishment of negative enrichment method using immunomagnetic beads to obtain high purity of tumor cells, the present study investigated the role of tumor cells and CD133＋ cell counting in the evaluation of disease and treatment efficacy in lung cancer patients with pleural effu- sion. Methods From 1 April, 2013 to 31 August, 2014, a total of 21 non-treated lung cancer patients with malignant pleural effusion and pathologically confirmed in respiratory department of clinical medical school of Yangzhou University were collected. All patients were treated with chemotherapy or gefitinib. Before treatment, 10 ml pleural effusion was collected. Tumor cells and CD133＋ cells from pleural effusion were counted using negative enrichment technology, chromo- some fluorescence in situ hybridization and fluorescent staining of CD133 antibody. The relationships between number of tumor cell and CD133＋ cell and the clinical significance were further analyzed. RESULTS In all the 21 patients, the medi an number of tumor cells and CD133＋ cells were 4 000/10 mL, and 19/10 mL respectively. There were no significant difference of tumor cell and CD133 ＋ cell numbers in different gender （x2 = 0. 597, P= 0. 550; ;x2= 0. 225, P= 0. 822, re- spectively）, age （x2 =0.533, P=0.594;x2=1. 288, P=0. 198, respectively） and pathological type （ x2=3.253, P =0. 197; ;x2=0. 034, P= 0. 983, respectively）. The numbers of tumor cell and CD133＋ cell were significantly higher in pa tients with pulmonary or distant lymph nodes or other organ metastasis than that in patients without or only with regional lymph node metastasis （ x2=4.398, P=0.036; X2 4. 605, P=0.032, respectively）. After treatment with 2 cycles of chemotherapy or 2 month gefitinib, there were 0 CR, 7 PR, 9 SD, 5 PD and the total response rate was 33.33%. The numbers of tumor cell and CD133＋ cell were significantly different between different treatment efficacy （x2= 7. 575, P= 0. 023 ; x2 = 7. 247, P =0. 027, respectively）. The better treatment efficacy patients got, the less the numbers of tumor cell and CD133＋ cell in PD patients were the highest. According to the median number of tumor and cell CD133＋ cell, all the patients were divided into three groups： low level of cell number （Both tumor cell and CD133＋ cell numbers were equal or less than median number）, medium level of cell number （tumor cells^median and CD133＋ cells ≤median, or CD133＋ cells 2〉median and tumor cells≤median） and high level of cell number （Both tumor cell and CD133＋ cell numbers were more than median number）. The incidence of pulmonary or distant lymph nodes or other organ metastasis in patients with moder- ate high level of cell number was significantly higher than that in patients with low level of cell number （x2 =12. 634, P= 0. 002）. Furthermore, the treatment efficacy in patients with low level of cell number was better than in patients with mod- erate-high level of cell number （x2 =10. 886, P =0.01）. CONCLUSION Tumor cell and CD133＋ cell counting in pleural effusion may be an effective predictor for disease evaluation and treatment efficacy in patients with advanced lung cancer.