The Anti-melanoma Effect Of Ethanolic Extract Of Alocasia Cucullata And Its Mechanism

BackgroundMalignant melanoma is a malignant tumor produced by dedifferentiation of melanocytes.It occurs mostly in the skin,accounting for 2%of the incidence of systemic malignancies,accounting for third of skin malignant tumors.Alocasia cucullata(Lour.)G.Don(Chinese tarao)(AC)belongs to the araceae family and usually applied to treat a variety of cancers in the folkrhizome,get a good therapeutic effect.At present,there is a lack of systematic research on the anti tumor effect of AC at home and abroad,and there is no report on the anti melanoma of AC.Objective:1.To evaluate the anti-melanoma(B16-F10/A375/A2058)effect of 50%ethanolic extract of AC(EAC)in vitro.2.To establish a melanoma model of C57BL/6 mice and study the effect of EAC on the tumor in vivo.3.To study the anti-tumor signaling pathways related to tumor in vivo and in vitro,clarify the mechanism of and the underlying mechanism of EAC,provide a theoretical basis for the folk medicine,and provide a new alternative drug for the treatment of melanoma.Research content:1.In vitro anti-tumor assay1.1 To detect the effect of EAC on the activity of B16-F10/A375/A2058 cells by MTT assay.1.2 To detect the effect of EAC the migration and invasion abilities of melanoma cells by wound healing and transwell invasion assays.1.3 To detect the expression of apoptosis related proteins in B16-F10 melanoma cells by western blot.2.In vivo assay2.1 To establish mouse melanoma model and detect the effect of EAC on the tumor and immune organs of the tumor bearing mice.2.2 To detect the histopathological changes in the tumor tissuses of mice bearing B16-F10 cells by HE staining.2.3 To detect the expression of apoptosis related proteins in the tumor tissues of mice by immunohistochemical.2.3 To detect the expression of apoptosis related proteins in the tumor tissues of mice by western blot.3.Protective mechanisms of EAC on spleen tissues in tumor-bearing mice3.1 To detect the histopathological changes in the spleen tissues of tumor-bearing mice by HE staining.3.2 To detect the apoptosis of spleen tissues of mice by Tunel assay.3.3 To detect the expression of apoptosis related proteins in the spleen tissues of mice by immunohistochemical assays3.4 To detect the effect of EAC on the activity of spleen lymphocyte in normal mice by CCK-8 assay.Result1.EAC inhibits proliferation,migration and invasion of melanoma cells in vitro.Western blot results showed that EAC can significantly up-regulated PTEN and down regulated p-PI3K and p-AKT protein expression in B16-F10 melanoma cells,but had no significant effect on PI3K and AKT protein expression compared with the control group.2.EAC inhibits the tumor growth of B16-F10 cells in vivo and its mechanism is related to the up-regulation of PTEN,down-regulation of p-PI3K and p-AKT protein expression.3.EAC can increase the spleen index of tumor bearing mice,inhibits the abnormal apoptosis of spleen tissue in tumor microenvironment,and promote the proliferation activity of lymphocytes of normal mice.Conclusion1.EAC can inhibit the proliferation,migration and invasion of melanoma cells in vitro and inhibit tumor growth in vivo.The mechanism may be related to the PTEN/PI3K/AKT signaling pathway.2.EAC has certain protective effect on spleen tissue of mice bearing tumor,and improves the activity of lymphocytes of normal mice.It is presumed that it has the function of enhancing immunity and anti-tumor.