中药天花粉对人恶性黑色素瘤细胞形态、增殖、周期和迁移的影响

Effect of Radix Trichosanthis Decoction on Cell Morphology,Cell Proliferation and Cell Migration in Human Melanoma A375 Cells

目的:研究和探讨天花粉水煎剂在体外对人黑色素瘤A375细胞的形态、增殖、周期和迁移的影响。方法:用显微镜观察天花粉水煎剂作用前后的A375细胞形态的改变;利用MTT法检测天花粉水煎剂对A375细胞生长和增殖的影响;采用流式细胞术检测天花粉水煎剂对A375细胞周期的影响;利用划痕实验观察天花粉水煎剂对A375细胞迁移的影响。结果:显微镜观察结果和MTT结果显示,利用不同质量浓度的天花粉水煎剂处理后,正常形态的细胞明显减少,细胞死亡数量增加;而且对A375细胞的增殖有不同程度的抑制,抑制率呈质量浓度依赖性。流式细胞术分析结果发现,随着质量浓度的增加,细胞增殖周期中G2期中细胞所占百分数与对照组相比,有统计学差异;细胞划痕实验结果显示,640和2 560μg/m L天花粉水煎剂组划痕愈合率低。结论:天花粉水煎剂在体外可以改变A375细胞的正常形态,导致细胞死亡;对A375细胞的增殖周期具有抑制作用,同时诱导细胞周期中的G2期阻滞;一定浓度的天花粉水煎剂对A375细胞的迁移具有抑制作用,可以有效地抑制黑色素瘤细胞的恶化和转移,有利于黑色素瘤的预后。

Objective：To investigate the effect of Radix Trichosanthis decoction on cell morphology, cell proliferation and cell migration in human melanoma A375 cells in vitro.Methods：Use the microscope and MTT method and the changes of the cellular morphology and cell proliferation were detected in A375 cells treated by Radix Trichosanthis decoction;flo wing cytometry was applied to detect the cell cycle in A375 cells;cell migration was evaluated by using scratch experi-ment and the effect of the cell migration was assessed by the scarification test.Results： Compared with the control group, we found that A375 cells treated by Radix Trichosanthis decoction had the lowest cell viability and the number of normal morphology cells was significantly reduced in A375 cells in concentration-dependent manner.A375 cells trea-ted with Radix Trichosanthis decoction showed that cell proliferation was more obviously suppressed than those untreated control using MTT assay.Flowing cytometry assay showed the percentage of G2 phase cells had a significant difference compared with the control group treated by different concentration Radix Trichosanthis decoction.The cell scratch analy-sis indicated A375 cells treated by 640 and 2 560μg/mL Radix Trichosanthis decoction had lower wound healing rate. Conclusion：We found Radix Trichosanthis decoction could cause cell death and cell morphology abnormal in vitro.We also found Radix Trichosanthis decoction could inhibit cell proliferation and cell migration in concentration-dependent manner.Interestingly, G2 phase arrest could be induced by Radix Trichosanthis decoction.Our findings can provide new knowledge about Radix Trichosanthis decoction in development of melanoma and indicate the potential application of Radix Trichosanthis decoction in the prognosis of melanoma.