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Quantitative Real-Time Reverse Transcription–PCR Study of the Expression of Vascular Endothelial Growth Factor (VEGF) Splice Variants and VEGF Receptors (VEGFR-1 and VEGFR-2) in Non–Small Cell Lung Cancer

¹ Laboratory of Analytical Chemistry, Department of Chemistry, University of Athens, Athens 15771, Greece.
² Sotiria General Hospital, Athens 11526, Greece.
³ Department of Pathology, Onassis Cardiac Surgery Center, Athens 17674, Greece.

^aAddress correspondence to this author at: Laboratory of Analytical Chemistry, Department of Chemistry, University of Athens, Athens 15771, Greece. Fax 30-210-7274750; e-mail lianidou{at}chem.uoa.gr.

Background: Vascular endothelial growth factor (VEGF) is a major regulator of angiogenesis and its expression is increased in non–small cell lung cancer (NSCLC). We aimed to determine the expression pattern of VEGF splice variants in NSCLC and its correlation with the clinicopathological characteristics of tumors.

Methods: We used real-time reverse transcription PCR to quantify the mRNA expression of total VEGF, 4 VEGF splice variants (VEGF₁₂₁, VEGF₁₆₅, VEGF₁₈₃, and VEGF₁₈₉), and 2 VEGF receptors (VEGFR-1 and VEGFR-2) in 27 pairs of cancerous and adjacent noncancerous tissues originating from patients with NSCLC.

Results: Total VEGF, VEGF₁₂₁, and VEGF₁₆₅ were expressed in all specimens, whereas VEGF₁₈₃ and VEGF₁₈₉ were present in small amounts in certain samples. Total VEGF, VEGF₁₂₁, and VEGF₁₆₅ mRNA was upregulated in cancerous compared with healthy tissues, whereas VEGF₁₈₃ and VEGF₁₈₉ expression tended to be higher in healthy tissues. The expression of VEGFRs was similar between matched specimens. No correlation was found between the expression of total VEGF or VEGF splice variants and the clinicopathological characteristics of tumors. The expression patterns of VEGF splice variants differed between tissue pairs. VEGF₁₂₁ was the major variant expressed in all samples; however, its relative expression was higher in cancerous tissues. The relative expression of VEGF₁₈₃ and VEGF₁₈₉ was upregulated in healthy lung tissues, whereas the ratio of VEGF₁₆₅ to total VEGF was similar between matched specimens.

Conclusions: The expression pattern of certain VEGF splice variants is altered during tumorigenesis. Our data support the hypothesis that during malignant progression an angiogenic switch favoring the shorter diffusible isoforms occurs.

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