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2-Methoxystypandrone inhibits signal transducer and activator of transcription 3 and nuclear factor-κB signaling by inhibiting Janus kinase 2 and IκB kinase.

Kuang S, Qi C, Liu J, Sun X, Zhang Q, Sima Z, Liu J, Li W, Yu Q - Cancer Sci. (2014)

Bottom Line: We have identified 2-methoxystypandrone (2-MS) from a traditional Chinese medicinal herb Polygonum cuspidatum as a novel dual inhibitor of JAK2 and IKK. 2-MS inhibits both interleukin-6-induced and constitutively-activated STAT3, as well as tumor necrosis factor-α-induced NF-κB activation. 2-MS specifically inhibits JAK and IKKβ kinase activities but has little effect on activities of other kinases tested.The inhibitory effects of 2-MS on STAT3 and NF-κB signaling can be eliminated by DTT or glutathione and can last for 4 h after a pulse treatment.We propose that the natural compound 2-MS, as a potent dual inhibitor of STAT3 and NF-κB pathways, is a promising anticancer drug candidate.

View Article: PubMed Central - PubMed

Affiliation: Division of Tumor Pharmacology, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Shanghai, China.

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Effects of 2-methoxystypandrone (2-MS) on Janus kinase 2 (JAK2) and activities of other human kinases. (a) HeLa cells were incubated with indicated concentrations of 2-MS for 2 h before stimulation with interleukin-6 (IL-6) (30 ng/mL) for 15 min. Whole cell lysates were processed for western blot analysis and probed with anti-P-JAK2 (Y1007/1008), anti-JAK2 or anti-GP130 antibodies. Anti-α-Tubulin antibody was used as a loading control. (b) HeLa cells were incubated with indicated concentrations of 2-MS for 2 h before being processed for western blot analysis. (c) In vitro kinase assay of JAK2. JAK2 protein immunoprecipitated from HeLa cells was subjected to in vitro kinase assay in the presence of indicated concentrations of 2-MS. (d) Effects of 2-MS on activities of human kinases. In vitro kinase assays were processed by Millipore Kinase Services. (e) In vitro kinase assay of IKKβ. IKKβ protein immunoprecipitated from HEK293 cells was subjected to in vitro kinase assay in the presence of indicated concentrations of 2-MS.
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fig03: Effects of 2-methoxystypandrone (2-MS) on Janus kinase 2 (JAK2) and activities of other human kinases. (a) HeLa cells were incubated with indicated concentrations of 2-MS for 2 h before stimulation with interleukin-6 (IL-6) (30 ng/mL) for 15 min. Whole cell lysates were processed for western blot analysis and probed with anti-P-JAK2 (Y1007/1008), anti-JAK2 or anti-GP130 antibodies. Anti-α-Tubulin antibody was used as a loading control. (b) HeLa cells were incubated with indicated concentrations of 2-MS for 2 h before being processed for western blot analysis. (c) In vitro kinase assay of JAK2. JAK2 protein immunoprecipitated from HeLa cells was subjected to in vitro kinase assay in the presence of indicated concentrations of 2-MS. (d) Effects of 2-MS on activities of human kinases. In vitro kinase assays were processed by Millipore Kinase Services. (e) In vitro kinase assay of IKKβ. IKKβ protein immunoprecipitated from HEK293 cells was subjected to in vitro kinase assay in the presence of indicated concentrations of 2-MS.

Mentions: The inhibitory effect of 2-MS on STAT3 activation suggested that 2-MS might interfere with the upstream components of the STAT3 signaling pathway. Because JAK2 is the major kinase to activate STAT3 while GP130 is important for tyrosine phosphorylation of JAK2 in the IL-6/STAT3 pathway,(22) we first analyzed the effects of 2-MS on the expression and phosphorylation of JAK2 and the expression of GP130. In HeLa cells, 10 μM 2-MS significantly inhibited phosphorylation of JAK2 (Y1007/1008), while it had no effects on the expressions of JAK2 and GP130 (Fig. 3a). In addition, 10 μM 2-MS significantly inhibited the phosphorylation of JAK2 (Y1007/1008) in HeLa cells without IL-6 stimulation, further suggesting that JAK2 is the direct target of 2-MS (Fig. 3b).


2-Methoxystypandrone inhibits signal transducer and activator of transcription 3 and nuclear factor-κB signaling by inhibiting Janus kinase 2 and IκB kinase.

Kuang S, Qi C, Liu J, Sun X, Zhang Q, Sima Z, Liu J, Li W, Yu Q - Cancer Sci. (2014)

Effects of 2-methoxystypandrone (2-MS) on Janus kinase 2 (JAK2) and activities of other human kinases. (a) HeLa cells were incubated with indicated concentrations of 2-MS for 2 h before stimulation with interleukin-6 (IL-6) (30 ng/mL) for 15 min. Whole cell lysates were processed for western blot analysis and probed with anti-P-JAK2 (Y1007/1008), anti-JAK2 or anti-GP130 antibodies. Anti-α-Tubulin antibody was used as a loading control. (b) HeLa cells were incubated with indicated concentrations of 2-MS for 2 h before being processed for western blot analysis. (c) In vitro kinase assay of JAK2. JAK2 protein immunoprecipitated from HeLa cells was subjected to in vitro kinase assay in the presence of indicated concentrations of 2-MS. (d) Effects of 2-MS on activities of human kinases. In vitro kinase assays were processed by Millipore Kinase Services. (e) In vitro kinase assay of IKKβ. IKKβ protein immunoprecipitated from HEK293 cells was subjected to in vitro kinase assay in the presence of indicated concentrations of 2-MS.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4317813&req=5

fig03: Effects of 2-methoxystypandrone (2-MS) on Janus kinase 2 (JAK2) and activities of other human kinases. (a) HeLa cells were incubated with indicated concentrations of 2-MS for 2 h before stimulation with interleukin-6 (IL-6) (30 ng/mL) for 15 min. Whole cell lysates were processed for western blot analysis and probed with anti-P-JAK2 (Y1007/1008), anti-JAK2 or anti-GP130 antibodies. Anti-α-Tubulin antibody was used as a loading control. (b) HeLa cells were incubated with indicated concentrations of 2-MS for 2 h before being processed for western blot analysis. (c) In vitro kinase assay of JAK2. JAK2 protein immunoprecipitated from HeLa cells was subjected to in vitro kinase assay in the presence of indicated concentrations of 2-MS. (d) Effects of 2-MS on activities of human kinases. In vitro kinase assays were processed by Millipore Kinase Services. (e) In vitro kinase assay of IKKβ. IKKβ protein immunoprecipitated from HEK293 cells was subjected to in vitro kinase assay in the presence of indicated concentrations of 2-MS.
Mentions: The inhibitory effect of 2-MS on STAT3 activation suggested that 2-MS might interfere with the upstream components of the STAT3 signaling pathway. Because JAK2 is the major kinase to activate STAT3 while GP130 is important for tyrosine phosphorylation of JAK2 in the IL-6/STAT3 pathway,(22) we first analyzed the effects of 2-MS on the expression and phosphorylation of JAK2 and the expression of GP130. In HeLa cells, 10 μM 2-MS significantly inhibited phosphorylation of JAK2 (Y1007/1008), while it had no effects on the expressions of JAK2 and GP130 (Fig. 3a). In addition, 10 μM 2-MS significantly inhibited the phosphorylation of JAK2 (Y1007/1008) in HeLa cells without IL-6 stimulation, further suggesting that JAK2 is the direct target of 2-MS (Fig. 3b).

Bottom Line: We have identified 2-methoxystypandrone (2-MS) from a traditional Chinese medicinal herb Polygonum cuspidatum as a novel dual inhibitor of JAK2 and IKK. 2-MS inhibits both interleukin-6-induced and constitutively-activated STAT3, as well as tumor necrosis factor-α-induced NF-κB activation. 2-MS specifically inhibits JAK and IKKβ kinase activities but has little effect on activities of other kinases tested.The inhibitory effects of 2-MS on STAT3 and NF-κB signaling can be eliminated by DTT or glutathione and can last for 4 h after a pulse treatment.We propose that the natural compound 2-MS, as a potent dual inhibitor of STAT3 and NF-κB pathways, is a promising anticancer drug candidate.

View Article: PubMed Central - PubMed

Affiliation: Division of Tumor Pharmacology, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Shanghai, China.

Show MeSH
Related in: MedlinePlus