Effect of antisense VEGF cDNA transfection on the growth of chronic myeloid leukemia K562 cells in vitro and in nude mice.
To further elucidate the role of vascular endothelial growth factor (VEGF) in the pathogenesis of chronic myeloid leukemia (CML), we transfected K562 cells with a VEGF(121)cDNA sense vector (S), an antisense (AS) vector or vector (V) alone. The growth of transfected cells was investigated by MTT and colony-formation assays, and apoptosis was measured by flow cytometry (FCM) of Annexin-V-FITC/PI dual labeled cells. Transfected cells were subcutaneously transplanted into nude mice and the microvessel density (MVD) of tumor masses was determined by vWF immunohistochemistry staining. We tested the supernatant of different transfected K562 cells against human bone marrow endothelial cells (BMECs), and examined the synergic effects of antisense VEGF(121)cDNA and IFNalpha or STI571 on the proliferation and apoptosis of K562 cells. We found that K562/AS transfectants exhibited a 49% reduction in VEGF secretion, whereas K562/S transfectants exhibited a 3-fold increase in VEGF secretion, all in comparison to the vector controls. K562 cells transfected with antisense VEGF(121)cDNA showed growth retardation in vitro. In transplanted nude mice in vivo, transfection of implanted cells with antisense VEGF(121)cDNA resulted in decreased tumor MVD, and increased apoptosis in the presence of IFNalpha. Taken together, these results suggest that VEGF may be involved in the pathogenesis of CML through autocrine and paracrine mechanisms, and that anti-VEGF therapy alone or in combination with conventional treatment may be beneficial for CML patients.
Ruan
Guo-Rui
GR
Institute of Hematology and People's Hospital, Peking University, 11 Xi-zhi-men South Street, Beijing 100044, China.
Liu
Yan-Rong
YR
Chen
Shan-Shan
SS
Fu
Jia-Yu
JY
Chang
Yan
Y
Qin
Ya-Zhen
YZ
Li
Jin-Lan
JL
Yu
Hong
H
Wang
Hui
H
D000818:Animals; D017209:Apoptosis; D002450:Cell Communication; D002455:Cell Division; D016373:DNA, Antisense / Q000235:genetics / Q000494:pharmacology; D018076:DNA, Complementary / Q000235:genetics; D006801:Humans; D016898:Interferon-alpha / Q000494:pharmacology; D020014:K562 Cells / Q000637:transplantation; D015464:Leukemia, Myelogenous, Chronic, BCR-ABL Positive / Q000209:etiology*; D051379:Mice; D008819:Mice, Nude; D009368:Neoplasm Transplantation; D014162:Transfection; D042461:Vascular Endothelial Growth Factor A / Q000235:genetics / Q000502:physiology*
D016373:DNA, Antisense; D018076:DNA, Complementary; D016898:Interferon-alpha; D042461:Vascular Endothelial Growth Factor A