Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 1997 Feb 4;94(3):979-84.
doi: 10.1073/pnas.94.3.979.

Vascular endothelial growth factor/vascular permeability factor is an autocrine growth factor for AIDS-Kaposi sarcoma

R Masood  1 , J Cai, T Zheng, D L Smith, Y Naidu, P S Gill
Affiliations

Vascular endothelial growth factor/vascular permeability factor is an autocrine growth factor for AIDS-Kaposi sarcoma

R Masood et al. Proc Natl Acad Sci U S A. .

Abstract

Kaposi sarcoma (KS) is the most common tumor associated with HIV-1 infection and develops in nearly 30% of cases. The principal features of this tumor are abnormal vascularization and the proliferation of endothelial cells and spindle (tumor) cells. KS-derived spindle cells induce vascular lesions and display enhanced vascular permeability when inoculated subcutaneously in the nude mouse. This finding suggests that angiogenesis and capillary permeability play a central role in the development and progression of KS. In this study, we show that AIDS-KS cell lines express higher levels of vascular endothelial growth factor/vascular permeability factor (VEGF/VGF) than either human umbilical vein endothelial cells or human aortic smooth muscle cells. AIDS-KS cells and primary tumor tissues also expressed high levels of Flt-1 and KDR, the receptors for VEGF, while the normal skin of the same patients did not show any expression. We further demonstrate that VEGF antisense oligonucleotides AS-1 and AS-3 specifically block VEGF mRNA and protein production and inhibit KS cell growth in a dose-dependent manner. Furthermore, growth of KS cells in nude mice was specifically inhibited by VEGF antisense oligonucleotides. These results show that VEGF is an autocrine growth factor for AIDS-KS cells. To our knowledge, this is the first report that shows that VEGF acts as a growth stimulator in a human tumor. Inhibitors of VEGF or its cognate receptors may thus be candidates for therapeutic intervention.

PubMed Disclaimer

Figures

Figure 1
Figure 1
Expression of VEGF mRNA in several AIDS–KS cells lines. (A) Total RNA (15 μg) from KSC10, KSC29, KSC13, KSC59, and KSY1 were electrophoresed, blotted, and hybridized to the human VEGF cDNA and β-actin probe. (B) Total RNA (15 μg) was isolated from KSC10, HUVEC, and AoSM and analyzed as in A. (C) Supernatants from equal numbers of cells from KSY1, KSC10, AoSM, HUVEC, and T1 were collected after 48 hr and analyzed for VEGF protein by ELISA.
Figure 2
Figure 2
Expression of Flt-1 and KDR mRNA in KSY1, HUVEC (HUVE), normal skin, and KS tumor tissue from an HIV+ patient, T1 (fibroblast), 23-1 (B lymphoma), and HUT-78 (T cell lymphoma). Equal amounts of RNA were reverse transcribed to generate cDNA. (A) cDNA was subjected to Flt-1- and KDR-specific PCR amplification (498- and 709-bp products, respectively) using paired primers as described. Lane M represents molecular weight markers. (B) cDNA from all samples were subjected to β-actin-specific PCR amplification (548-bp product).
Figure 3
Figure 3
Effect of VEGF antisense oligonucleotides on cell growth. (A) KS cells were treated with VEGF antisense AS-1, AS-2, AS-3, AS-4, AS-5, and scrambled oligonucleotides at concentrations of 5 μM. Cell proliferation was measured on day 5. (B) AIDS–KS cells were treated with VEGF AS-1, AS-3, and scrambled oligonucleotides at concentrations ranging from 1 to 10 μM; cell proliferation was measured on day 5. Several control cell lines were also treated with VEGF AS-1 oligonucleotides (C), VEGF AS-3 oligonucleotides (D), and VEGF scrambled oligonucleotides (E). Cell proliferation was measured on day 5, as described earlier. Data from all above experiments represent the mean ± SD of two separate experiments performed in quadruplicate. (F) Effect of rhVEGF on the growth of KS and HUVEC cells. Cells were seeded at 1 × 104 cells per well in 24 plates and treated with rhVEGF (1 and 10 ng/ml) for 48 hr. Cell counts were performed, and the results represent the mean ± SD of an experiment performed in quadruplicate. (G) rhVEGF abrogates the effect of VEGF antisense on AIDS–KS cell growth. KS cells were seeded at a density of 1 × 104 cells per well in 24-well plates. Cells were treated with 1 and 10 μM of AS-1 or AS-3 alone or with rhVEGF (10 ng/ml) on day 1 and day 2. Cell proliferation was measured after72 hr. The data represent the mean ± SD of two experiments performed in quadruplicate.
Figure 4
Figure 4
Effect of VEGF antisense (AS-1 and AS-3) and scramble (S) oligonucleotides on VEGF expression. Total RNA was isolated from AIDS–KS cells treated with various concentrations of AS-1 (A), AS-3 (B), and S (C). Total RNA was reverse transcribed to generate cDNA. PCR was carried out for VEGF and β-actin. (Upper) PCR products of 535 bp and 403 bp corresponding to VEGF121 and VEGF165 mRNA species of VEGF. (Lower) The 548-bp PCR product of β-actin. (NT, no treatment; M, molecular size marker.) The numbers 25-41 and 18-33 represent PCR cycles. (D) The supernatants of KS cells treated with AS-3 and scrambled VEGF antisense oligonucleotide were also collected at 48 hr, and VEGF protein was quantitated by ELISA. The results represent the mean ± SD of two separate experiments done in duplicate.
Figure 5
Figure 5
Effect on tumor growth of VEGF antisense oligonucleotides in nude mice. KSY1 cells (107) were inoculated subcutaneously in the lower back of BALB/c Nu+/NU+ athymic mice. AS-1, AS-3, and scrambled (S) VEGF oligonucleotides and diluent (PBS) were injected i.p. daily for 5 days (day 2–6). Mice were sacrificed on day 14 and tumor size was measured. Data represent the mean ± SD of 10 mice in each group.
See this image and copyright information in PMC

Similar articles

See all similar articles

Cited by

See all "Cited by" articles

References

    1. Safai B, Johnson K G, Myskowski P L, Cunnungham-Randles S, Godbold J H, Dupont B. Ann Intern Med. 1985;103:744–750. - PubMed
    1. Haverkos H W, Drotman D P, Morgan M. N Engl J Med. 1985;312:1518–1523. - PubMed
    1. Ruszczak Z, Silva A M, Orfanos C E. Am J Dermatopathol. 1987;9:388–398. - PubMed
    1. Regezi S A, Macphail L A, Daniels T E, De Souza Y G, Greenspan J S, Greenspan D. Am J Pathol. 1993;143:240–249. - PMC - PubMed
    1. Kaplan L D, Hopewell P C, Jaffe H, Goodman P C, Bottles K, Volberding P A. J Acquired Immune Defic Syndr. 1988;1:23–30. - PubMed

Publication types

MeSH terms

Substances