Tubocapsanolide A inhibits transforming growth factor-beta-activating kinase 1 to suppress NF-kappaB-induced CCR7.
Withanolides are C(28) steroidal lactones isolated from plants that exhibit potent anti-cancer activity. The chemokine receptor CCR7 is important for lymphatic invasion of cancer cells and is overexpressed in metastatic breast cancer cells. A bioactive withanolide tubocapsanolide A (Tubo A) suppressed NF-kappaB-mediated CCR7 expression in breast cancer cells and attenuated their migration toward lymphatic endothelial cells. Chromatin immunoprecipitation assay confirmed that binding of NF-kappaBto the consensus site localized at the -398/-389 of human CCR7 promoter was repressed by Tubo A. Tubo A inhibited IkappaB kinase (IKK) and p38 kinase and downstream mitogen and stress-activated protein kinase 1 (MSK1) activity to reduce IkappaB degradation and to suppress NF-kappaB activation. Co-expression of IKK and MSK1 fully rescued Tubo A-induced inhibition. In addition, ectopic expression of transforming growth factor-beta-activating kinase (TAK1), the common upstream kinase of IKK and MSK1, also completely reversed the inhibition by Tubo A. Most importantly, Tubo A reduced NF-kappaB activation, CCR7 expression, and lymph node metastasis of breast cancer in vivo. We conclude that Tubo A inhibits TAK1 to repress NF-kappaB-induced CCR7 expression in breast cancer cells and suggest that Tubo A may be useful for the prevention of lymphatic invasion of breast cancer cells.
Pan
Mei-Ren
MR
Graduate Institute of Medicine, College of Medicine, Kaohsiung Medical University, Kaohsiung 807.
Chang
Hui-Chiu
HC
Graduate Institute of Medicine, College of Medicine, Kaohsiung Medical University, Kaohsiung 807.
Wu
Yang-Chang
YC
Graduate Institute of Natural Products, Kaohsiung Medical University, Kaohsiung 807; National Sun Yat-Sen University-Kaohsiung Medical University Joint Research Center, Kaohsiung 804.
Huang
Chao-Cheng
CC
Department of Pathology, Chang Gung Memorial Hospital-Kaohsiung Medical Center, Chang Gung University College of Medicine, Kaohsiung 833.
Hung
Wen-Chun
WC
National Sun Yat-Sen University-Kaohsiung Medical University Joint Research Center, Kaohsiung 804; Institute of Biomedical Sciences, National Sun Yat-Sen University, 70, Lien-Hai Road, Kaohsiung 804; Center for Gene Regulation and Signal Transduction Research, National Cheng Kung University, Tainan 701, Taiwan. Electronic address: hung1228@ms10.hinet.net.
D001483:Base Sequence; D045744:Cell Line, Tumor; D047369:Chromatin Immunoprecipitation; D017931:DNA Primers; D004875:Ergosterol / Q000031:analogs & derivatives* / Q000494:pharmacology; D005434:Flow Cytometry; D006801:Humans; D020930:MAP Kinase Kinase Kinases / Q000037:antagonists & inhibitors*; D016328:NF-kappa B / Q000037:antagonists & inhibitors* / Q000235:genetics / Q000502:physiology; D011401:Promoter Regions, Genetic; D054400:Receptors, CCR7 / Q000037:antagonists & inhibitors* / Q000502:physiology; D020133:Reverse Transcriptase Polymerase Chain Reaction; D000099151:MAP Kinase Kinase Kinase 7
D017931:DNA Primers; D004875:Ergosterol; D020930:MAP Kinase Kinase Kinases; D016328:NF-kappa B; D054400:Receptors, CCR7; D000099151:MAP Kinase Kinase Kinase 7; C515370:CCR7 protein, human; C522061:tubocapsanolide A