黄连素靶向调控TLR4信号通路对恶性黑色素瘤生物学行为的影响及机制研究

目的 探讨黄连素对恶性黑色素瘤Toll样受体4(TLR4)信号通路的调节作用,评估其对脂多糖(LPS)诱导的恶性黑色素瘤A375细胞增殖、迁移、凋亡的影响,为防治恶性黑色素瘤提供新的药物(黄连素)治疗依据.方法 采用Western blotting法检测不同浓度黄连素对A375细胞TLR4、核因子κB (NF-κB)表...

Effect and mechanism of berberine-targeted regulation of TLR4 signaling pathway on the biological behaviour of malignant melanoma

Objective To explore the regulatory effect of berberine on the TLR4 signaling pathway in malignant melanoma,to evaluate its effect on the proliferation,migration,and apoptosis of malignant melanoma A375 cells induced by lipopolysaccharide(LPS),and to provide a new drug(berberine)treatment basis for the prevention and treatment of malignant melanoma.Methods Western blotting was used to detect the intervention effects of different concentrations of berberine on the expression of TLR4 and NF-κB in A375 cells.The experimental groups were as follows:group A was control group,group B was LPS group,group C was TAK-242 group,group D was LPS+TAK-242 group,group E was LPS+TAK-242+80μmol/L berberine group,and group F was LPS+TAK-242+120μmol/L Berberine group.CCK-8,Transwell and flow cytometry assays were used to detect the effects of berberine targeting TLR4 on the proliferation,migration,and apoptosis of A375.We used enzyme-linked immunosorbent assay to detect the expression of IL-10 and TGF-βin the supernatant of each group of cell influences.Results With increased concentration of berberine,the expression of TLR4 and NF-κB protein was down-regulated more significantly(all P<0.05).Compared with group A,the proliferation and migration ability of group B was enhanced,but no difference existed in apoptosis.The proliferation and migration ability of group C was weakened,and apoptosis was increased.The proliferation and migration ability of group D was weaker than that of group B and stronger than that of group C.Compared with group D,the proliferation and migration ability of E and F groups weakened,and apoptosis increased.At high concentrations of berberine,the cell proliferation and migration ability weakened more obviously(all P<0.05).The expression levels of IL-10 and TGF-βin group B increased(all P<0.05),the expression level of IL-10 in group C decreased,and the expression of TGF-βdid not change significantly.The IL-10 of group D was lower than that of group B and increased compared with that of group C.The expression levels of IL-10 and TGF-βin group F were significantly reduced(all P<0.05).Conclusion Berberine inhibits the proliferation and migration of LPS-induced malignant melanoma cells and promotes their apoptosis by inhibiting the TLR4/NF-κB signaling pathway.Berberine may be used as a medicine to treat malignant melanoma.