Blood, 1 November 2000, Vol. 96, No. 9, pp. 3181-3187
NEOPLASIA
In vitro and in vivo production of vascular endothelial growth
factor by chronic lymphocytic leukemia cells
Haijuan Chen,
Andy T. Treweeke,
Dave C. West,
Kathleen J. Till,
John C. Cawley,
Mirko Zuzel, and
Cheng H. Toh
From the Departments of Haematology and Immunology,
University of Liverpool, Liverpool, United Kingdom.
Expansion of primary solid tumors and their malignant
dissemination are angiogenesis-dependent. Vascular endothelial growth factor (VEGF) is the key factor playing a pivotal role in solid tumor-induced angiogenesis. Recent studies indicate that angiogenesis may also be involved in the pathogenesis of certain hemic malignancies, including B-cell chronic lymphocytic leukemia (B-CLL). Mechanisms underlying angiogenesis in B-CLL and the role of VEGF in this process
are incompletely understood. In this study, it was examined whether
angiogenically functional VEGF is produced by B-CLL cells. Immunohistochemical staining with antibodies against VEGF and CD34, an
endothelial cell marker, demonstrated the presence of VEGF protein and
abundant blood vessels in infiltrated lymphoreticular tissues. Low
levels of VEGF were detected by ELISA in the culture media of
unstimulated cells; this was enhanced up to 7-fold by hypoxic
stimulation. SDS-PAGE and Western blot analysis of the concentrated
culture media showed 2 isoforms of VEGF protein with molecular weights
of 28 and 42 kd, respectively. RNA hybridization showed that these
cells expressed VEGF mRNA. Reverse transcription-polymerase chain reaction, combined with nucleotide sequence analysis, revealed that the predominantly expressed isoforms were VEGF121 and VEGF165. Moreover, 3H-thymidine incorporation and an in vivo
angiogenic assay demonstrated that the VEGF produced by CLL cells can
induce angiogenesis by stimulating endothelial cell proliferation. In
conclusion, this study shows that B-CLL cells produce VEGF and
demonstrates the angiogenic effects of this growth factor, which may be
relevant for the tissue phase of the disease.
