白山毛桃根提取物抑制黑色素瘤M21细胞增殖作用及其机制研究

Research on Effect and Potential Mechanism of Extractive of Radix Actnidiae Erianthae on Proliferation Inhibition in Human Melanoma M21 Cells

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 investigate the effects of the extractive of Radix Actnidiae Erianthae on the proliferation of human melanoma M21 ceils and to further explore its molecular mechanism. Methods：The inhibitory rate of cell growth was measured by MTF; the cell invasion was detected by wound healing assay;Real- Time PCR was used to detect Program death ligand 1 （PD- L1 ） and PD - L2 genes expressions, and Western blot analysis was used to detect PD - LI and PD - L2 protein expressions. Re- sults：In the same conditions, the extractive of Radix Actnidiae Erianthae had significant activity in inhibiting M21 cells, showing a dose - time dependency （P 〈 0. 05 ）, but had no effect of inhibition on normal epithelial pigment RPE cells （P 〈 0. 05 ） ;the extract can inhibit the M21cells invasion ;the mRNA and protein expressions of PD -L1 and PD -L2 were decreased in M21 cells. Con- clusion：The extractive of Radix Actnidiae Erianthae could significantly inhibit the proliferation and the cell invasion in human melanoma M21 cells ;the mechanisms may be related to reducing the mRNA and protein expression of PD -L1 and PD -L2 in M21 ceils.