| Investigating the biological mechanisms underlying melanoma progression and searching for effective drugs are critical for developing effective therapies against melanoma. The bioactive ingredients from Chinese herbal medicines are considered to be abundant sources for developing new anti-tumor drugs. Sesquiterpene lactones are a large class of active component in medicinal plants, widely found in acanthaceae, umbelliferae, euphorbiaceae, magnoliaceae, Mmenispermaceae, asteraceae and other plants. Extracts only from the asteraceae of sesquiterpene lactones contains more than3000structures Parthenolide, helenalin, triptolide, costunolide, eupatolide and other sesquiterpene lactones have been reported to have anti-inflammatory and cytotoxic activities. we researched on Xanthatin which extracted from Xanthium strumarium L. Performed the X-crystal diffraction and DEPT,1H,1H-COSY, NOESY, HSQC and HMBC experiments to identified the structures. We screened a variety of tumor cell lines such as lang cancer cell A549, hepatocellular carcinoma cell HepG2, hepatocellular carcinoma cell HuH7, melanoma cell B16-F10and found that xanthatin had obvious effect on promoting apoptosis of melanoma cells B16-F10. MTS、Annexin-V/PI double staining、Flow cytometric analysis are applied to detect anticancer activity of Xanthatin. MTS experiments showed that treatment with xanthatin at10μM for24h induced about69.35%inhibition of cell proliferation, and the value of IC50was7.36μM. Xanthatin inhibited B16-F10cell growth in dose-/time-dependent manners. Further study was implemented. Using Western blot method to detecting expression of proteins in Wnt/β-catenin signaling pathway. After treatment with xanthatin, the non-canonical WNT-5A was down-regulated, and nuclear β-catenin was up-regulated, indicating that Wnt/β-catenin signaling pathway could be activated by xanthatin. Xanthatin had obvious effect on promoting apoptosis of melanoma cells B16-F10, so we established the in vivo anti-tumor model. Because the cell line B16-F10is mouse source, so we choosed C57BL/6mice to build up the in vivo melanoma model. Histopathological examination immunohistochemistry、 immunofluorescence experiments are applied to further investigate the in vivo anti-tumor target protein expression. Our data showed that the inhibitory effect of xanthatin at high dose (0.4mg/10g) was similar to that of taxol, and the inhibition ratio of them were42.57%and41.22%respectively, which were significantly different from the model group (**P<0.01,**P<0.01). From the histopathological analysis, some necrosis signs were especially prominent in sections as shown in the pink color around the blood vessels in taxol and xanthatin groups, but the normal tumor cells were purple. Based on these findings, we postulated that xanthatin might affect vascular endothelial growth factor and microvessels and reduce the supply of the the energy to inhibit tumor growth, Additional experiment provided evidence for this postulation. Less PCNA-positive and survivin-positive cells with brown in xanthatin and taxol groups were observed. Cell proliferative marker PCNA.Apply AB5500mass spectrometry to investigate xanthatin pharmacokinetic parameters in vivo, the results show after the intravenous injection of Xanthatin in low, middle and high dose groups (2.4mg/200g4.8mg/200g9.6mg/200g), the mean Cmax are418.72±137.51,904.89±193.53,1773.46±1733.1ng/mL respectively. The dynamical behavior of the animals has a linear characteristic, Cmax and AUC0-t showed a dose-dependent growth trend in the dose range of the experimental.From this experiment, we were delighted to find that xanthatin had excellent anti-tumor activity. It is likely to be developed into antineoplastic in the future. |
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