Received: 16 February 2017
Accepted: 1 May 2017
DOI: 10.1002/pro6.16
ORIGINAL ARTICLE
Clinical significance of tumor cells and CD133+ cells from malignant pleural effusion in patients with lung cancer Chen Yong1 Li Jun1
Ling-feng Min2
Xi-zhi Zhang1
Jun-jun Yang2
Wen-jing Xu2
Jia-rong Bian2
Xing-xiang Xu2
1 Department of Medical Oncology, Northern
Jiangsu Peopleβs Hospital, Clinical Medical College of Yangzhou University, Yangzhou, Jiangsu, China 2 Department of Respiratory Medicine, Northern
Jiangsu Peopleβs Hospital, Clinical Medical College of Yangzhou University, Yangzhou, Jiangsu, China Correspondence Xing-xiang Xu, Department of Respiratory Medicine, Northern Jiangsu Peopleβs Hospital, Clinical Medical College of Yangzhou University, Yangzhou, Jiangsu, China. Email:
[email protected] Funding Information Clinical Medical Science and Technology Project of Jiangsu Province, Grant/Award Number: SBL201230144; National Natural Science Foundation of China, Grant/Award Number: 81302015; Natural Science Foundation of Yangzhou City, Grant/Award Number: YZ2014044; Jiangsu Government Scholarship
Abstract Objective: The composition of malignant pleural effusion in lung cancer is so complex that it is very difficult to obtain high-purity tumor cells from pleural effusions. Based on the establishment of the negative enrichment method using immunomagnetic beads to obtain high-purity tumor cells, the present study investigated the clinical significance of tumor cells and CD133+ cells in lung cancer patients with malignant pleural effusion. Methods: From 1 April 2013 to 31 August 2014, 21 untreated lung cancer patients with pathologically confirmed malignant pleural effusions were enrolled in a study from the Respiratory Department of Clinical Medical School of Yangzhou University. All patients were treated with chemotherapy or gefitinib. Before treatment, 10 mL of pleural effusion was collected. Tumor cells and CD133+ cells from 10 mL pleural effusion were counted by using negative enrichment technology, chromosomal fluorescence in situ hybridization and fluorescent staining with anti-CD133 antibody. In addition, the relationships between the numbers of tumor cells and CD133+ cells, and clinical characteristics were further analyzed. Results: Among the 21 patients, the median number of tumor cells and CD133+ cells was 4000/10 mL and 19/10 mL, respectively. There were no significant differences in the numbers of tumor cells and CD133+ cells in patients of different gender (π 2 = 0.597, P = 0.550 and π 2 = 0.225, P = 0.822, respectively), age (π 2 = 0.533, P = 0.594 and π 2 = 1.288, P = 0.198, respectively) and different pathological types of lung cancer (π 2 = 3.253, P = 0.197 and π 2 = 0.034, P = 0.983, respectively). The numbers of tumor cells and CD133+ cells were significantly higher in patients with pulmonary, distant lymph node or other organ metastasis than in patients without distant metastasis or with only regional lymph node metastasis (π 2 = 4.398, P = 0.036 and π 2 = 4.605, P = 0.032, respectively). After treatment with two cycles of chemotherapy or gefitinib for 2 months, 0 patients had complete response, seven had partial response, nine had stable disease, five had progressive disease, and the total response rate was 33.33%. The numbers of tumor cells and CD133+ cells were significantly different between the patients with different treatment efficacies (π 2 = 7.575, P = 0.023 and π 2 = 7.247, P = 0.027, respectively). The numbers of tumor cells and CD133+ cells in patients with partial response were the lowest, whereas the numbers of tumor cells and CD133+ cells in patients with progressive disease were the highest. The patients were divided into three groups according to the median number of tumor cells and CD133+ cells: low cell numbers (both tumor cell and CD133+ cell numbers were equal to or less than the median number), moderate cell numbers (tumor cells > median and CD133+ cells β€ median or CD133+ cells > median and tumor cells β€ median) and high cell numbers (both tumor cell and CD133+ cell numbers were more than the median numbers). The incidence of pulmonary, distant lymph node or
This is an open access article under the terms of the Creative Commons Attribution-NonCommercial-NoDerivs License, which permits use and distribution in any medium, provided the original work is properly cited, the use is non-commercial and no modifications or adaptations are made. c 2017 The Authors. Precision Radiation Oncology published by John Wiley & Sons Australia, Ltd on behalf of Shandong Cancer Hospital & Institute. Prec. Radiat. Oncol. 2017;1:58β63.
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other organ metastasis in patients with a moderate-to-high cell numbers was significantly higher than that in patients with a low cell numbers (π 2 = 12.634, P = 0.002). Furthermore, the treatment efficacy in patients with a low cell number was better than that in patients with a moderate-to-high cell numbers (π 2 = 10.886, P = 0.01). Conclusion: Tumor cells and CD133+ cells in pleural effusions might be used as an effective predictor of disease progression and treatment efficacy in patients with advanced lung cancer. KEYWORDS
CD133, efficacy prediction, lung cancer, negative enrichment, pleural effusion
1
INTRODUCTION
2.2
Treatment
Pleural effusion is a common complication in lung cancer, and approx-
The patients were treated with chemotherapy or gefitinib. Four
imately 76% of malignant pleural metastasis arises from lung cancer.
patients with small cell lung cancer were treated with chemother-
Nearly 15% of newly diagnosed lung cancer patients have pleural
apy with etoposide + cisplatin/carboplatin. Among the four patients
effusion, and 40β50% of lung cancer patients will have pleural effusion
with squamous cell carcinoma, three patients were treated with the
during disease progression. Malignant pleural effusion has a complex
paclitaxel + cisplatin/carboplatin regimen, and one patient received
composition, containing a small number of tumor cells and cancer
the gemcitabine + cisplatin regimen. Among the 13 patients with ade-
In pleural effusions, lymphocytes account for
nocarcinoma, two patients were treated with the paclitaxel + cis-
50β70% of all nucleated cells. At present, it is difficult to obtain
platin/carboplatin regimen, and seven patients were treated with
high-purity tumor cells by conventional centrifugation and density
the pemetrexed + cisplatin/carboplatin regimen. One patient was
gradient centrifugation, so the study of pleural effusion is still pri-
sequentially treated with single-agent pemetrexed chemotherapy and
marily limited to diagnosis. In a previous study, our group developed
one with docetaxel chemotherapy. In addition, two patients were
a negative immunofluorescence enrichment method (also called
treated with gefitinib based on epidermal growth factor receptor test
subextraction technic method) that can separate most white blood
results. Platinum drugs including cisplatin (Chinese Medicine Zhunzi
cells from malignant pleural effusion, and enrich for high-purity tumor
H20010743; Jiangsu Hansoh Pharmaceutical Group Co., Ltd, Lianyun-
cells in lung cancer.3
This method improved the positive rate of clinical
gang, China) were given at a total dose of 75 mg/m2 in a single dose
diagnosis, and helped to further explore the clinical significance of
or divided into three doses; the dose of carboplatin (Chinese Medicine
pleural effusion. In the present study, immunostaining fluorescence
Zhunzi H20020181; Qilu Pharmaceutical Group, Jinan, China) was cal-
in situ hybridization (FISH) and immunofluorescence staining were
culated according to the area under the curve of concentration of
stem cells
(CSCs).1,2
CD133+
cells from malignant pleural
unbound carboplatin in the plasma versus time. The area under the
effusions, and to further explore the clinical significance of counting
curve value was 5, and carboplatin was given at a single dose. The
tumor cells and CD133+ cells.
combination of drugs included gemcitabine (1000 mg/m2 , d1, d8; Chi-
used to identify tumor cells and
nese Medicine Zhunzi HZ20030104; Hansoh Pharmaceutical Group Co., Ltd, Lianyungang, China), paclitaxel (175 mg/m2 , d1; Chinese
2
METHODS
Medicine Zhunzi H20058719; Yangtze River Pharmaceutical Group, Taizhou, China), docetaxel (75 mg/m2 , d1; Chinese Medicine Zhunzi
2.1
Patients
A total of 21 newly diagnosed lung cancer patients with malignant pleural effusion were enrolled in the present study. Among them, there were 14 men and seven women. Nine patients were aged
