Study On The Anti-tumor Activity And Mechanism Of HDF,a Farrerol Derivative,with A Mouse Model

6,8-dimethyl-7,4 ’-dimethoxy-5-hydroxyl methane(HDF),a derivative of farrerol,is a patented compound of our group with independent intellectual property rights.Previous studies have shown that HDF exhibits microtubule-targeted and antitumor activity in vitro.On this basis,we further investigated the antitumor activity of HDF with a mouse model in vivo.In this study,we established a subcutaneous xenograft model of melanoma B16 cells in mice,and evaluated the tumor inhibition effect of HDF by intraperitoneal injection.Furthermore,the possible anti-tumor mechanisms of HDF were further explored in vitro using He La cells as a cell model.Materials and methods: 1.The tumor volume and tumor weight were measured to evaluate the inhibitory effect of HDF on the subcutaneous transplanted tumor in mice.The effect of HDF on the basic physical signs of mice was evaluated by measuring the weight of body and main organs of mice.The expressions of Ki67,CD31,Bax and Bcl-2 in mouse tumor tissues were evaluated by immunohistochemical assay.TUNEL assay was used to detect the apoptosis of tumor cells in mice.The toxicity of HDF to normal tissues and organs of mice was investigated by HE staining of main organ slices.2.The effects of HDF on the expressions of various proteins in cells were studied by proteomic techniques and bioinformatics analysis using He La cells as a model in vitro.Western-blotting assay was used to verify the effect of HDF on the expression of proteins in the NF-κB/Bcl-2 apoptosis pathway.The effect of HDF on Bax and Bcl-2 at the transcription levels were studied by q-PCR assay.Results: 1.HDF exhibited significant inhibitory effect on tumor growth in mice.Immunohistochemical results of Ki67 and CD31 showed that HDF could inhibit tumor cell proliferation and angiogenesis in Kunming mice.TUNEL assay demonstrated that HDF can promote tumor cell apoptosis in vivo.Immunohistochemical results of Bcl-2 and Bax indicated that HDF could promote apoptosis by regulating the expression of Bcl-2 and Bax proteins in vivo.There was no significant effect of HDF on body weight and organ coefficient of mice.HE staining of main organs of mice showed that HDF has a certain of toxicity to liver and kidney of mice,while no significant toxicity to other organs was observed.2.Results of proteomic techniques and bioinformatics analysis showed that HDF can affect the NF-κB signaling pathway and the expression of Bcl-2 family.Western-blotting assay proved that HDF can promote apoptosis by affecting the NF-κB signaling pathway and the content of Bcl-2/Bax protein,and q-PCR results demonstrated that HDF can regulate the expression of Bcl-2/Bax at the transcriptional level.Conclusion: HDF exhibits a promising anti-tumor effect in vivo that can inhibit tumor growth significantly in mice.The anti-tumor mechanisms of HDF include inhibition of tumor cell proliferation,tumor metastasis,angiogenesis and promotion of tumor cell apoptosis by regulating the NF-κB/Bcl-2 apoptosis pathway.