| In Vitro | Kinase Assay: Honokiol is a bioactive, biphenolic phytochemical that possesses potent antioxidative, anti-inflammatory, antiangiogenic, and anticancer activities by targeting a variety of signaling molecules. It inhibits the activation of Akt and enhances the phosphorylation of ERK1/ERK2.
Cell Assay: Honokiol shows pro-apoptotic effects in melanoma, sarcoma, myeloma, leukemia, bladder, lung, prostate, oral squamous cell carcinoma and colon cancer cell lines. Honokiol is effective on inducing apoptosis in SVR angiosarcoma cells. Treatment of SVR cells with honokiol causes decreased phosphorylation of MAP kinase, akt, and c-src. In addition, honokiol potentiates TRAIL-mediated apoptosis, and honokiol cytotoxicity is partially abrogated by neutralizing antibodies to TRAIL. Honokiol also has direct antiangiogenic activity, in that honokiol blocks the phosphorylation and rac activation due to VEGF-VEGFR2 interactions. Honokiol causes apoptosis in CLL cells through activation of caspase 8, followed by caspase 9 and 3 activation. Honokiol prevents interleukin-4-mediated survival of CLL cells, and potentiats the cytotoxicity of chlorambucil, fludarabine, and cladribine. Honokiol kills myeloma cells from relapsed patients at doses that does not kill PBMCs. Caspase 3, 7, 8, and 9 are induced by honokiol treatment, as well as PARP cleavage. Honokiol is found to induce apoptosis in the colon cancer cell lines RKO. Honokiol potentiates apoptosis, suppresses osteoclastogenesis, and inhibits invasion through modulation of nuclear factor-kappaB activation pathway. Honokiol may act as a potent anti-inflammatory agent with multipotential activities due to an inhibitory effect on the PI3K/Akt pathway. |
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| Animal model | MAmalian: For anticancer in vivo studies, the MDA-MB-231 cells (2 million) are injected into mammary fat tissue. Two weeks after the tumor cell injections, palpable tumors are observed in mammary tissues, which is an indication of tumor formation. Then drug treatment either in free form or in nanomicellar forms is given orally at the dose of 40 and 80 mg/kg daily. The drug treatment is continued for 4 weeks, and the tumor volumes and body weights are recorded weekly. After 4 weeks of treatment, animals are sacrificed; final tumor volumes and weights are measured. These tumors are used for western blot and immunohistochemical analysis. For western blot experiments, tumor tissues are stored at –80°C till the analysis is done. For IHC, tumors are fixed in formal saline. |
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