Wortmannin reduces metastasis and angiogenesis of human breast cancer cells via nuclear factor-κB-dependent matrix metalloproteinase-9 and interleukin-8 pathways.
OBJECTIVE
To investigate whether inhibition of Akt phosphorylation by the phosphatidylinositol 3-kinase (PI3K) inhibitor, wortmannin, reduces metastasis and angiogenesis in a human breast cancer cell line via nuclear factor (NF)-κB-dependent matrix metalloproteinase (MMP)-9 and interleukin (IL)-8 pathways.


METHODS
MDA-MB-231 cells were treated with wortmannin 0-200 nM for 4 h. Restoration of Akt activity was evaluated by transfection of cells with constitutively active myristoylated Akt (myr-Akt). NF-κB, MMP-9 and IL-8 proteins were detected by electrophoretic mobility shift assay, Western blot or enzyme-linked immunosorbent assay. The chicken embryo chorio-allantoic membrane assay, cell motility and migration assays were used to evaluate angiogenesis and invasion in vitro. A mouse pseudo metastatic breast cancer model was used to assess the effects of wortmannin on metastasis and angiogenesis in vivo.


RESULTS
Wortmannin inhibited the phosphorylation of Akt, upregulation of NF-κB, MMP-9, IL-8, and in vitro cell invasion and angiogenesis, in a dose-dependent manner. Transfection of myr-Akt reversed the cellular and biochemical effects of wortmannin in vitro. Wortmannin also significantly inhibited tumour metastasis and angiogenesis in vivo.


CONCLUSIONS
The findings of the present study suggest that wortmannin inhibits metastasis and angiogenesis in breast cancer cells via PI3K/Akt/NF-κB-mediated MMP-9 and IL-8 signalling pathways.
Centre of Breast Disease, Affiliated Hospital of Qingdao University Medical College, Qingdao University, Qingdao, Shandong Province, China.
Jianguo
J
Li
J
J
Li
Funian
F
Li
F
F
Li
Haibo
H
Wang
H
H
Wang
Xingang
X
Wang
X
X
Wang
Yao
Y
Jiang
Y
Y
Jiang
Dongxiao
D
Li
D
D
Li
D000730:Androstadienes / Q000494:pharmacology / Q000627:therapeutic use*; D000818:Animals; D001943:Breast Neoplasms / Q000201:enzymology / Q000378:metabolism / Q000473:pathology*; D045744:Cell Line, Tumor; D024202:Electrophoretic Mobility Shift Assay; D005260:Female; D006801:Humans; D016209:Interleukin-8 / Q000378:metabolism*; D020780:Matrix Metalloproteinase 9 / Q000378:metabolism*; D051379:Mice; D016328:NF-kappa B / Q000378:metabolism*; D009362:Neoplasm Metastasis / Q000188:drug therapy*; D009389:Neovascularization, Pathologic / Q000188:drug therapy*; D010766:Phosphorylation; D051057:Proto-Oncogene Proteins c-akt / Q000378:metabolism; D000077191:Wortmannin
D000730:Androstadienes; D016209:Interleukin-8; D016328:NF-kappa B; D051057:Proto-Oncogene Proteins c-akt; D020780:Matrix Metalloproteinase 9; D000077191:Wortmannin