[Frontiers in Bioscience E2, 411-423, January 1, 2010]

[Frontiers in Bioscience E2, 411-423, January 1, 2010]

Expression of VEGF, VEGF-C and VEGFR-2 in in situ and invasive SCC of cervix

Robert Jach¹, Joanna Dulinska-Litewka², Piotr Laidler², Andrzej Szczudrawa¹, Andrzej Kopera¹, Łukasz Szczudlik¹, Michał Pawlik¹, Krzysztof Zając¹, Monika Mak¹, Antoni Basta¹

1Chair of Gynecology and Obstetrics, Department of Gynecology, Obstetrics and Oncology; ² Medical Biochemistry,Jagiellonian University, Medical College, Krakow, Poland

TABLE OF CONTENTS

1. Abstract
2. Introduction
3. Materials and Methods: 3.1. Patients and sampling; 3.2. Tissues; 3.3. Immunohistochemistry; 3.4. RNA extraction; 3.5. cDNA synthesis and RT-PCR analysis; 3.6. cDNA synthesis for real-time PCR; 3.7. Protein determination; 3.8. Western blot analysis; 3.9. Statistical analysis
4. Results: 4.1. Immunostaining results for VEGFR-2 and LV(D2-40) in pre-invasive and invasive cervical lesions; 4.2. Proteins determination - mRNA and protein
5. Discussion
6. Acknowledgments
7. References

1. ABSTRACT

Cervical squamous cell carcinoma (SCC) arises from the metaplastic epithelium and develops slowly through dysplastic changes (i.e., cervical intraepithelial neoplasia - CIN) to carcinoma in situ and invasive cancer. There is little data concerning the quantitation of vascular endothelial growth factor (VEGF) and its correlation to the clinical or pathologic characteristics of SCC. This study assessed the expression of VEGF, VEGF-C and their receptor VEGFR-2 in 35 samples of normal cervical tissue, 35 - CIN1, 35 - CIN2 (25 non-pregnant, 15 pregnant women), 35 - CIN3 and 30- SCC. VEGF, VEGF-C and VEGFR-2 were analyzed using RT-PCR, RQ-PCR, immunohistochemical staining and Western blot. VEGF, VEGF-C and VEGFR-2 were not detected in normal cervical epithelium. In CIN and SCC, both forms of VEGF and its receptor were identified, indicating a correlation between the increasing expression and staging of carcinoma. Results show the important role of VEGF in cervical progression and that the switch to the lymphangiogenesis phenotype occurs prior to the stage of invasion likely at CIN2/3.