[Amphiregulin antisense RNA expression inhibits angiogenesis of human breast cancer in nude mice].
OBJECTIVE
To investigate the anti-angiogenic effect of amphiregulin (AR) antisense RNA expression in breast cancer.


METHODS
Human AR cDNA antisense plasmid was transfected into NS2T2A1 cells (a human breast cancer cell line). Two selected clones expressed AR antisense RNA (AR AS1 and AR AS3 cell lines) in which AR protein expression was reduced. Control cell line NS2T2A1 V was obtained by empty vector transfection. These cells were injected subcutaneously into nude mice. The effects of conditioned media on proliferation of human microvascular endothelial cells (HMEC) were evaluated and VEGF secreted by the cells was measured by ELISA method. In tumor tissues, VEGF expression levels were measured by quantitative RT-PCR, and CD31-immunostaining was used for intra-tumoral vascular quantification.


RESULTS
The proliferation index of HMEC cells grown in conditioned media with AR AS1 and AR AS3 was significantly reduced in comparison with that of control cells, accompanied by a decreased VEGF secretion. In tumors derived from AR AS1 and AR AS3 cells, intra-tumoral vascularization was reduced to about 50% of that derived from control cell line, accompanied with a decrease of VEGF expression.


CONCLUSIONS
Amphiregulin antisense RNA expression inhibits efficiently the angiogenesis in breast cancer, suggesting this growth factor could represent a novel therapeutic target in breast cancer.
Ma
Lin
L
Department of Radiotherapy, Chinese PLA General Hospital, Beijing 100853, China.
Serova
Maria
M
Podgorniak
Marie Pierre
MP
Berthois
Yolande
Y
Mourah
Samia
S
Calvo
Fabien
F
D000230:Adenocarcinoma / Q000098:blood supply / Q000473:pathology; D066258:Amphiregulin; D020533:Angiogenesis Inhibitors / Q000096:biosynthesis* / Q000235:genetics; D000818:Animals; D001943:Breast Neoplasms / Q000098:blood supply* / Q000473:pathology; D066255:EGF Family of Proteins; D006023:Glycoproteins / Q000096:biosynthesis* / Q000235:genetics; D006801:Humans; D036341:Intercellular Signaling Peptides and Proteins / Q000096:biosynthesis* / Q000235:genetics; D051379:Mice; D008819:Mice, Nude; D009389:Neovascularization, Pathologic; D016372:RNA, Antisense / Q000096:biosynthesis* / Q000235:genetics; D042442:Vascular Endothelial Growth Factors / Q000037:antagonists & inhibitors
C587848:AREG protein, human; D066258:Amphiregulin; D020533:Angiogenesis Inhibitors; C587849:Areg protein, mouse; D066255:EGF Family of Proteins; D006023:Glycoproteins; D036341:Intercellular Signaling Peptides and Proteins; D016372:RNA, Antisense; D042442:Vascular Endothelial Growth Factors