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目的:研究白山毛桃根提取物对黑色素瘤M21细胞增殖影响,并进一步探讨其作用潜在的分子机制。方法:MTT法检测白山毛桃根提取物对M21细胞增殖抑制作用;细胞划痕实验检测M21细胞侵袭迁移力;采用Real-Time PCR及Western blot方法检测M21细胞上免疫共刺激分子Program death ligand 1(PD-L1)、PD-L2基因及蛋白表达变化。结果:白山毛桃根提取物可以抑制M21细胞增殖,有统计学意义(P<0.05),呈时间和浓度依赖性;而在同条件下对人正常上皮色素RPE细胞无抑制作用,无统计学差异(P>0.05);其次该药能抑制M21细胞侵袭迁移能力及明显下调M21细胞上PD-L1、PD-L2基因及蛋白表达。结论:白山毛桃根提取物能抑制M21细胞增殖、侵袭迁移能力,其机制可能是通过下调与肿瘤细胞免疫逃逸密切相关的PD-L1、PD-L2分子基因和蛋白表达而实现的。
OBJECTIVE: To study the effect of Baimayutang root extract on the proliferation of melanoma M21 cells and to explore its potential molecular mechanism. Methods: MTT assay was used to detect the proliferation of M21 cells induced by P. dauricus root extract. Cell scratch assay was used to detect the invasion and migration of M21 cells. Real-time PCR and Western blot were used to detect the expression of immune-stimulatory molecules Program death ligand 1 ( PD-L1), PD-L2 gene and protein expression changes. Results: Baimao peach root extract could inhibit the proliferation of M21 cells with statistical significance (P <0.05) in a time and concentration-dependent manner; while under the same conditions, there was no significant difference in the inhibitory effect on human normal epithelial RPE cells (P> 0.05). Second, the drug could inhibit the invasion and migration of M21 cells and down-regulate the expression of PD-L1 and PD-L2 gene and protein in M21 cells. Conclusion: Baimayama root extract can inhibit the proliferation, invasion and migration of M21 cells. The mechanism may be through down-regulating the gene and protein expression of PD-L1 and PD-L2 molecules that are closely related to the tumor cell immune escape.