Human malignant melanoma is a very aggressive and highly angiogenesis-dependent tumor. Basic fibroblast growth factor and angiogenin are the potentially important angiogenic factors for melanoma progression and metastasis. Many studies have mainly focused on how they induce angiogenesis. In the present study, we investigated the effects of basic fibroblast growth factor on the expression of angiogenin and melanoma cell growth.
Methods and study design
Angiogenin mRNA and protein expression were investigated by means of semi-quantitative reverse transcriptase polymerase chain reaction assay and western blotting. We analyzed cell proliferation using MTT, flow cytometry and soft agar assay. Immunofluorescence staining was applied to investigate co-localization and nuclear translocation.
Results
We found that basic fibroblast growth factor negatively affected the expression of angiogenin in A375 cells. The result showed that down-regulation of basic fibroblast growth factor induced decreased cell proliferation of A375 cells, and in basic fibroblast growth factor up-regulated cells, cell proliferation was increased. We demonstrated that basic fibroblast growth factor protein was co-localized with angiogenin and that it underwent nuclear translocation in A375 cells.
Conclusion
These findings suggest that there is a cooperation mechanism between basic fibroblast growth factor and angiogenin in A375 cells, and the cooperation mechanism affects the progress of tumor cell proliferation and angiogenesis. Free full text available at www.tumorionline.it
MoscatelliD., PrestaM., RifkinD.B.: Purification of a factor from human placenta that stimulates endothelial cell protease production, DNA synthesis and migration.Proc Natl Acad Sci USA, 83: 2091–2095, 1986.
KleinS., BikfalviA., BirkenmeierT.M., GiancottiF.G., RifkinD.B.: Integrin regulation by endogenous expression of 18-kDa fibroblast growth factor-2.J Biol Chem, 271: 22583–22590, 1996.
8.
BasilicoC., MoscatelliD.: The FGF family of growth factors and oncogenes. Adv.Cancer Res, 59: 115–165, 1992.
9.
BeckerD., MeierC.B., HerlynM.: Proliferation of human malignant melanomas is inhibited by antisense oligodeoxynucleotides targeted against basic fibroblast growth factor.EMBO J, 8: 3685–3691, 1989.
10.
WangY.P., BerkerD.: Antisense targeting of basic fibroblast growth factor and fibroblast growth factor receptor-1 in human melanomas blocks intratumoral angiogenesis and tumor growth.Nat Med, 3: 887–893, 1997.
11.
FettJ.W., StrydomD.J., LobbR.R., AldermanE.M., BethuneJ.L., RiordanJ.F., ValleeB.L.: Isolation and characterization of angiogenin, an angiogenic protein from human carcinoma cells.Biochemistry, 24: 5480–5486, 1985.
12.
OlsonK.A., ByersH.R., KeyM.E., FettJ.W.: Prevention of human prostate tumor metastasis in athymic mice by antisense targeting of human angiogenin.Clin Cancer Res, 7: 3598–3605, 2001.
13.
OlsonK.A., ByersH.R., KeyM.E., FettJ.W.: Inhibition of prostate carcinoma establishment and metastatic growth in mice by an antiangiogenin monoclonal antibody.Int J Cancer, 98: 923–929, 2002.
14.
OlsonK.A., FettJ.W., FrenchT.C., KeyM.E., ValleeB.L.: Angiogenin antagonists prevent tumor growth in vivo.Proc Natl Acad Sci USA, 92: 442–446, 1995.
15.
HartmannA., KunzM., KostlinS., GillitzerR., ToksoyA., BröckerE., KleinC.: Hypoxia-induced up-regulation of angiogenin in human malignant melanoma.Cancer Res, 59: 1578–1583, 1999.
16.
MonteroS., GuzmanC., Cortes-FunesH., ColomerR.: Angiogenin expression and prognosis in primary breast carcinoma.Clin Cancer Res, 4: 2161–2168, 1998.
17.
ChopraV., DinhT.V., HanniganE.V.: Circulating serum levels of cytokines and angiogenic factors in patients with cervical cancer.Cancer Invest, 16: 152–159, 1998.
18.
LiD., BellJ., BrownA., BerryC.L.: The observation of angiogenin and basic fibroblast growth factor gene expression in human colonic adenocarcinomas, gastric adenocarcinomas, and hepatocellular carcinomas.J Pathol, 172: 171–175, 1994.
19.
ShimoyamaS., ShimizuN., TsujiE., YamasakiK., KawaharaM., KaminishiM.: Distribution of angiogenin and its gene message in colorectal cancer patients and their clinical relevance.Anticancer Res, 22: 1045–1052, 2002.
20.
ChopraV., DinhT.V., HanniganE.V.: Serum levels of interleukins, growth factors and angiogenin in patients with endometrial cancer.J Cancer Res Clin Oncol, 123: 167–172, 1997.
21.
ShimoyamaS., KaminishiM.: Increased angiogenin expression in gastric cancer correlated with cancer progression.J Cancer Res Clin Oncol, 126: 468–474, 2000.
22.
HisaiH., KatoJ., KobuneM., MurakamiT., MiyanishiK., TakahashiM., YoshizakiN., TakimotoR., TeruiT., NiitsuY.: Increased expression of angiogenin in hepatocellular carcinoma in correlation with tumor vascularity.Clin Cancer Res, 9: 4852–4859, 2003.
KatonaT.M., NeubauerB.L., IversenP.W., ZhangS., BaldridgeL.A., ChengL.: Elevated expression of angiogenin in prostate cancer and its precursors.Clin Cancer Res, 11: 8358–8363, 2005.
25.
OlsonK.A., FrenchT.C., ValleeB.L., FettJ.W.: A monoclonal antibody to human angiogenin suppresses tumor growth in athymic mice.Cancer Res, 54: 4576–4579, 1994.
26.
KishimotoK., LiuS., TsujiT., OlsonK.A., HuGF: Endogenous angiogenin in endothelial cells is a general requirement for cell proliferation and angiogenesis.Oncogene, 22: 445–456, 2004.
27.
SongJ., WangJ., YangJ., JiangC., ShenW., WangL.: Influence of angiogenin on the growth of A375 human melanoma cells and the expression of basic fibroblast growth factor.Melanoma Res, 6: 119–126, 2006.
28.
YangJ., WangJ., ZhaoJ., ZuoD., LiX., WangL.: Influence of basic fibroblast growth factor on the growth of HeLa cells and the expression of angiogenin.Oncol Rep, 21: 949–955, 2009.
29.
WangJ., YangJ., YuanD., WangJ., ZhaoJ., WangL.: Effects of basic fibroblast growth factor on angiogenin expression and cell proliferation of H7402 human hepatoma cells.J Genet Genom, 36: 399–407, 2009.
30.
DorkinT.J., RobinsonM.C., MarshC., NealD.E., LeungH.Y.: aFGF immunoreactivity in prostate cancer and its co-localization with bFGF and FGF8.J Pathol, 189: 564–569, 1999.
31.
LiuX., ZhengS.X., ZhouL.J., ZhuX.Z.: Basic fibroblast growth factor up-regulates the expression of vascular endothelial growth factor in primary cultured rat astrocytes.Acta Pharmacol Sin, 21: 19–22, 2000.
32.
RabieA.B., LuM.: Basic fibroblast growth factor up-regulates the expression of vascular endothelial growth factor during healing of allogeneic bone graft.Arch Oral Biol, 49: 1025–1033, 2004.
XuZ.P., MontiD.M., HuG.F.: Angiogenin activates human umbilical artery smooth muscle cells.Biochem Biophys Res Commun, 285: 909–914, 2001.
35.
WidmannC., GibsonS., JarpeM.B.: Mitogen activated protein kinase: conservation of a three-kinase module from yeast to human.Physiol Rev, 79: 143–180, 1999.
36.
KinM., SataM., UenoT., TorimuraT., InuzukaS., TsujiR., SujakuK., SakamotoM., SugawaraH., TamakiS., TanikawaK.: Basic fibroblast growth factor regulates proliferation and motility of human hepatoma cells by an autocrine mechanism.J Hepatol, 27: 677–687, 1997.
37.
ShengZ., ChangS.B., ChiricoW.J.: Expression and purification of a biologically active basic fibroblast growth factor fusion protein.Protein Expr Purif, 27: 267–271, 2003.
PedersonT.: Growth factors in the nucleolus?J Cell Biol, 143: 279–281, 1998.
40.
ClausP., DoringF., GringelS., OstermeyerF.M., FuhlrottJ., KraftT., GrotheC.: Differential intranuclear localization of fibroblast growth factor-2 isoformsand specific interaction with the survival of motoneuron protein.J Biol Chem, 278: 479–485, 2003.
41.
XuZ.P., TsujiT., RiordanJ.F., HuG.F.: Identification and characterization of an angiogenin-binding DNA sequence that stimulates luciferase reporter gene expression.Biochemistry, 42: 121–128, 2003.
42.
ShengZ., LewisJ.A., ChiricoW.J.: Nuclear and nucleolar localization of 18 kDa fibroblast growth factor-2 is controlled by C-terminal signals.J Biol Chem, 279: 40153–40160, 2004.
