雷公藤甲素对黑色素瘤A375细胞EMT通路及侵袭能力的作用研究

Effects of triptolide on epithelial-mesenchymal transition and invasion of melanoma A375 cells

目的研究雷公藤甲素对人黑色素瘤细胞A375侵袭转移的影响及其分子机制。方法采用CCK-8实验检测黑色素瘤细胞的增殖;细胞侵袭实验观察雷公藤甲素对黑色素瘤细胞运动力和侵袭力的影响;Western blot检测黑色素瘤细胞相关蛋白HIF-1α及EMT相关蛋白的表达。结果与对照组比较,雷公藤甲素可显著抑制A375细胞的增殖活性,差异有统计学意义(P<0.05)。Transwell小室实验结果显示,雷公藤甲素可抑制黑色素瘤细胞的侵袭转移能力;Western blot结果显示,雷公藤甲素作用A375细胞后HIF-1α表达显著下调,同时HIF-1α表达量与雷公藤甲素具有浓度依赖性。与对照组比较,EMT通路相关蛋白E-cadherin表达下调、N-cadherin和Vimentin表达上调(P<0.01)。结论雷公藤甲素能抑制人黑色素瘤A375细胞增殖,可能通过HIF-1α/EMT途径抑制黑色素瘤A375细胞侵袭转移。

Objective To explore the effects and molecular mechanisms of triptolide on the invasion and metastasis of human melanoma A375 cells.Methods CCK-8 test was used to detect the cell proliferation.The cell invasion test was used to observe the effects of triptolide on the motility and invasiveness of melanoma cells.Western blot was used to detect the expression of HIF-1αprotein and EMT-related proteins in melanoma cells.Results Compared with the control group,triptolide significantly inhibited the proliferation of A375 cells,with statistically significant difference(P<0.05).The results of Transwell experiment showed that triptolide could inhibit the invasion and metastasis of melanoma cells.The result of Western blot showed that the expression of HIF-1αin A375 cells was significantly down-regulated after treatment with triptolide,in a concentration-dependent manner.Compared with the control group,the expression of E-cadherin was down-regulated and the expressions of N-cadherin and Vimentin were up-regulated(P<0.01).Conclusion Triptolide can inhibit the proliferation of human melanoma A375 cells,which may inhibit the invasion and metastasis of melanoma A375 cells through HIF-1α/EMT pathway.