Multiplex PCR-Based Detection of Circulating Tumor Cells in Lung Cancer Patients Using CK19, PTHrP, and LUNX Specific Primers

Autor: Michael Koutsilieris, Konstantinos Syrigos, Haralampos Maragos, Anastasia Katseli, Adrianos Nezos
Rok vydání: 2013
Předmět:
Adult
Male
Pulmonary and Respiratory Medicine
Cancer Research
Lung Neoplasms
Adenocarcinoma
Real-Time Polymerase Chain Reaction
chemistry.chemical_compound
Circulating tumor cell
Carcinoma
Non-Small-Cell Lung
Molecular marker
Multiplex polymerase chain reaction
Biomarkers
Tumor
Tumor Cells
Cultured
medicine
Humans
RNA
Messenger
Neoplasm Metastasis
Lung cancer
Lymph node
Aged
Glycoproteins
Neoplasm Staging
Aged
80 and over
Keratin-19
A549 cell
Messenger RNA
Reverse Transcriptase Polymerase Chain Reaction
business.industry
Parathyroid Hormone-Related Protein
Bone metastasis
Middle Aged
Neoplastic Cells
Circulating
Phosphoproteins
Prognosis
medicine.disease
Small Cell Lung Carcinoma
Molecular biology
medicine.anatomical_structure
Oncology
chemistry
Case-Control Studies
Carcinoma
Squamous Cell
Carcinoma
Large Cell
Female
business
Multiplex Polymerase Chain Reaction
Follow-Up Studies
Zdroj: Clinical Lung Cancer. 14:513-520
ISSN: 1525-7304
DOI: 10.1016/j.cllc.2013.04.007
Popis: Introduction The aim of this study was to develop a multiplex polymerase chain reaction (PCR)-based method for detection of circulating tumor cells in peripheral blood of lung cancer (LC) patients. Patients and Methods Peripheral blood was collected from 71 healthy donors and 125 LC patients at different pathological stages. Samples were analyzed using multiplex PCR, and specific primers for CK19, PTHrP, and LUNX mRNA. The sensitivity of our method was set at 10 LC cells (A549 cells) in 3 mL of peripheral blood of healthy donors using spiking experiments. Results The detection rates in LC patients for CK19, PTHrP, and LUNX were 45.6%, 64.8%, and 28%, and in healthy individuals were 7%, 7%, and 5.6%, respectively. Overall, our method produced 77.8% positive detections for at least 1 molecular marker. Twenty-eight (22.2%) were negative for expression of all markers, 39 (31.2%) were positive for expression of 1 marker, 42 (33.6%) were positive for expression of 2 markers, and 17 (13.6%) were positive for expression of all 3 markers. Detection of CK19 mRNA expression positively correlated with LC stage and distant metastases. PTHrP mRNA detection correlated positively with LC stage, presence of bone metastasis, and squamous cell carcinoma, and LUNX mRNA detection correlated with lymph node involvement. Combined detection of 2 or 3 markers was significantly correlated with metastatic disease, and negative detection of all 3 molecular markers was correlated with early stage nonmetastatic disease. Conclusion Multiple PCR-based detection of CK19, PTHrP, and LUNX mRNA expression provides useful information for disease stage and dissemination in LC patients.
Databáze: OpenAIRE
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