No full-text available
To read the full-text of this research,
you can request a copy directly from the authors.
PO-18 - Fibronectin EDA/EDB is expressed in adherent SCLC NCI-H69 cells and in pleural effusions of lung cancer patients: possible implication for drug resistance
Abstract
Introduction
Small cell lung cancer (SCLC) is an extremely aggressive tumour which metastasizes early. Even if chemotherapy can achieve an initial regression, relapses due to chemo-resistance are almost inevitable. Sethi et al (Nat Med. 1999;5:662-668) reported that matrix proteins are essentially involved in the development of drug resistance. SCLC cells in suspension culture secrete negligible amounts of matrix proteins
Aim
For a more detailed study of the SCLC ability to produce matrix proteins we applied a recently introduced cell culture model of adherence selected SCLC (Salge et al. J Cancer Res Clin Oncol. 2001;127(2):139-411) and analysed pleural effusions form lung cancer patients.
Materials and Methods
Adherent cells were selected from the SCLC cell line NCI-H69 after exposure to cellular stress. Pleural effusion were obtained from lung cancer patients (SCLC and NSCLC) and from pleural effusions (PE) with congestive heart failure Protein expression was analysed by western blotting (WB) and flow cytometry using specific antibodies against the fibronectin extra domain A (FnEDA) and B (FnEDB) (Sirius, Italy), and for integrins alpha 1-5 and beta 1-3. Drug resistance was assessed with the metabolic stain MTT (3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium-bromid).
Results
SCLC suspension cells expressed negligible amounts of fibronectin. In contrast, adherent H69 cells, which showed a significantly reduced chemo-sensitivity against carboplatin and doxorubicin, strongly expressed FnEDA and to a lesser extent FnEDB. Furthermore, in adherent cells expression of various integrins was up-regulated, in particular integrins alpha5/beta3, representing potential binding sites for FnEDA/FnEDB. Analysis of pleural effusions clearly showed the presence of FnEDA/ FnEDB in those of lung cancer patients, whereas in benign pleural effusion almost no FnEDA/ FnEDB was found.
Conclusions
Our data reveal the presence of Fn, and its splice variants FnEDA/EDB in particular, in adherent SCLC cells as well as in malignant PE. We assume that the splice variants FnEDA/ FnEDB are linked to cancer progression and chemo-resistance in this tumour type.
... For these techniques, the The extra domain B splice variant of bronectin (EDB-FN) is an extracellular matrix (ECM) protein deposited by tumor-associated broblasts and is associated with tumor growth, angiogenesis, and invasion [7,8]. EDB-FN, as a tumor-associated ECM protein, is overexpressed in breast cancer [9,10], prostate cancer [11,12], small cell lung cancer [13], and colorectal cancer [14]. An EDB-FN-targeted peptide, Thr-Val-Arg-Thr-Ser-Ala-Asp (ZD2), was initially reported by Lu et al [12]. ...
Purpose
The Breast Imaging Reporting and Data System (BI-RADS) and 2-deoxy-2-[¹⁸F]fluoro-D-glucose ([¹⁸F]F-FDG) positron emission tomography (PET) have some limitations in the diagnosis of breast cancer. The extra domain B splice variant of fibronectin (EDB-FN), which is overexpressed in several cancer types, including breast cancer, is an approved diagnostic and therapeutic target of EDB-FN-positive cancers. Herein, we provide the EDB-FN-targeted peptide EDBp as a noninvasive oncological imaging modality for the molecular imaging of breast cancer.
Procedures
The binding of the peptide EDBp-Cy5 to EDB-positive breast cancer cells was investigated by flow cytometry and confocal microscopy. Near-Infrared Fluorescence (NIRF) imaging was performed at 48 hours after 1 µmol EDBp-Cy5 was intravenously injected into subcutaneous MDA-MB-231 breast cancer model mice or pulmonary metastasis model mice. Model mice were intravenously injected with approximately 100 µCi of [¹⁸F]-EDBp, and PET/CT scanning was performed after 2 hours of in vivo circulation.
Results
Our results demonstrate that EDBp-Cy5 had a strong binding ability to the MDA-MB-231 cells. EDBp-Cy5 exhibited specific tumor accumulation in MDA-MB-231 subcutaneous and pulmonary metastasis model mice. Importantly, we proposed and validated an EDBp peptide-based radiotracer [¹⁸F]-EDBp for PET imaging. [¹⁸F]-EDBp provided excellent diagnostic value for PET imaging of breast cancer, especially in subcutaneous tumors. The uptake of [¹⁸F]-EDBp in subcutaneous tumors (6.53 ± 0.89%, ID/g) was unexpectedly higher than that in the kidney (4.96 ± 0.20, %ID/g), which is the main metabolic organ of peptides. In the pulmonary metastasis model mice, PET/CT imaging showed that [¹⁸F]-EDBp moderately accumulated in the pulmonary metastases.
Conclusion
The high tumor uptake of this radiotracer in mice suggests its potential for application in PET imaging of EDB-FN-positive breast cancer for disease staging of regional and distant metastases and relapse monitoring.
The extra domain B splice variant of fibronectin (EDB-FN), which is overexpressed in several cancers, is an approved diagnostic and therapeutic target of cancers. The aim of this study was to evaluate the EDB-FN-targeting peptide EDBp as a noninvasive imaging modality for molecular imaging of breast cancer in mice. Western blot, flow cytometry and immunofluorescence were used to assess the expression level of EDB-FN and its binding to EDRp in MCF7, SKBR3, 4T1, EMT6, MDA-MB-231 and MDA-MB-453 cells. Establishment MDA-MB-231-luc cells-based subcutaneous tumor model mice or pulmonary metastasis model mice. The EDRp molecular probes to perform fluorescent probes for near-infrared fluorescence (NIRF)·and PET imaging of model mice. Our results demonstrate that EDBp-Cy5 had a strong binding ability to the MDA-MB-231 cells and exhibited specific tumor accumulation in MDA-MB-231 subcutaneous and pulmonary metastasis model mice. Importantly, the EDBp peptide-based radiotracer [¹⁸F]-AlF-NOTA-EDBp provided excellent diagnostic value for positron emission tomography (PET) imaging of breast cancer, especially in subcutaneous model mice. The uptake of [¹⁸F]-AlF-NOTA-EDBp in subcutaneous tumors (6.53 ± 0.89%, ID/g) was unexpectedly higher than that in the kidney (4.96 ± 0.20, %ID/g). The high tumor uptake of these probes in mice suggests their potential for application in imaging of EDB-FN-positive breast cancer for disease staging of regional and distant metastases.
Cancer is the second leading cause of death worldwide, despite the many treatments available, cancer patients face side effects that reduce their quality of life. Therefore, there is a need to develop novel strategies to increase the efficacy of treatments. In this study, gold nanoparticles obtained by green synthesis with Coffea arabica green bean extract were loaded with Doxorubicin, (a highly effective but non-specific drug) by direct interaction and using commercial organic ligands that allow colloidal dispersion at physiological and tumor pH. Conjugation of these components resulted in stable nanohybrids at physiological pH and a tumor pH release dependent, with a particle size less than 40 nm despite having the ligands and Doxorubicin loaded on their surface, which gave them greater specificity and cytotoxicity in H69 tumor cells.
ResearchGate has not been able to resolve any references for this publication.