木鳖子单体化合物对羟基桂皮醛诱导小鼠黑素瘤B16细胞的分化及其机制

发布时间：2018-05-29 06:36

本文选题：对羟基桂皮醛 + 木鳖子　；参考：《中国肿瘤生物治疗杂志》2014年03期

【摘要】：目的:探讨从中药木鳖子中提取的单体化合物对羟基桂皮醛(p-hydroxylcinnamaldehyde,PHD)对小鼠黑素瘤B16细胞分化的影响及其可能的作用机制。方法:以10、20、40μmol/L PHD作用B16细胞,设空白对照及福司克林(Forskelin)阳性对照组。磺酰罗丹明(suphrhodamineB,SRB)法检测PHD对B16细胞生长的抑制率,平板克隆集落形成实验检测细胞的克隆形成能力,Giemsa染色观察细胞形态,比色法检测细胞中黑色素含量和酪氨酸酶(tyrosinase,Tyr)活性,划痕愈合实验检测细胞的迁移能力,Western blotting检测Tyr、酪氨酸酶相关蛋白1(tyrosinase protein,Trp1)及MAPK信号转导通路中p-P38、pJNK和p-ERK1/2的表达。结果:PHD对黑素瘤B16细胞具有明显的增殖抑制作用,10、20、40μmol/L PHD作用B16细胞48h后,其增殖抑制率与对照组相比明显增加[(12.78±0.87)%、(37.70±2.28)%、(42.17±4.18)%vs 0,P0.05],20、40μmol/L PHD组增殖抑制率与Forskelin组相比明显增加[(37.70±2.28)%、(42.17±4.18)%vs(22.00±1.13)%,P0.05]。40μmol/L PHD处理B16细胞24、48、72 h后,B16细胞呈典型分化形态,黑色素含量及Tyr活性与对照组相比均明显增加[(0.097±0.02)、(0.13±0.04)、(0.15±0.05)vs(0.085±0.003);(1.11±0.31)、(1.43±0.05)、(1.67±0.49)vs(0.64±0.10),P0.05];细胞集落形成能力和迁移能力都明显降低(均P0.05)。与空白对照组相比,PHD处理组细胞Tyr和Trp1的表达明显增加(P0.05),MAPK信号通路中p-P38和p-JNK的表达水平也明显增加(P0.05),而p-ERK活性差异则无统计学意义(P0.05)。结论:PHD通过上调MAPK信号通路中p-P38和p-JNK的活性而对小鼠黑素瘤B16细胞产生明显的增殖抑制,并在形态和功能上诱导黑素瘤细胞的分化。
[Abstract]:Objective: to investigate the effect of monomer compound p-hydroxylcinnamaldehyde( PHD) extracted from Trionyx chinensis L. on the differentiation of mouse melanoma B16 cells and its possible mechanism. Methods: B16 cells were treated with 10 渭 mol/L PHD and 20 渭 mol/L PHD respectively. The cells were divided into blank control group and Forskelin positive control group. The inhibitory rate of PHD on the growth of B16 cells was detected by suprahrhodamine Bsrb method. The colony forming ability of B16 cells was detected by plate colony forming assay. The melanin content and tyrosinase Tyr activity in B16 cells were detected by colorimetric assay, and the cell morphology was observed by Giemsa staining. The migration ability of cells was detected by scratch healing assay. Tyr, tyrosinase associated protein 1(tyrosinase protein 1 (TRP 1) and the expression of p-P38 pJNK and p-ERK1/2 in MAPK signal transduction pathway were detected by Western blotting. Results [WT5HZ] PhD had a significant inhibitory effect on the proliferation of melanoma B16 cells. After 48 hours of treatment, the proliferation of B16 cells was inhibited by 10 ~ (20) ~ (20) ~ 40 渭 mol/L PHD. Compared with the control group, the inhibitory rate of proliferation was significantly increased [12.78 卤0.87 卤37.70 卤2.28 卤42.17 卤4.18)%vs 0 4.18)%vs] the inhibitory rate of proliferation in the 40 渭 mol/L PHD group was significantly higher than that in the Forskelin group [37.70 卤2.28 4.18)%vs(22.00 卤1.13P0.05] .40 渭 mol/L PHD treated B16 cells for 24 h and 48 h after treatment, the proliferation inhibition rate of the B16 cells showed typical differentiation morphology, and the proliferation inhibition rate of the 40 渭 mol/L PHD group was significantly higher than that of the Forskelin group (P < 0.05). Compared with the control group, the content of melanin and the activity of Tyr were significantly increased [0.13 卤0.04 卤0.13 卤0.03 0.05)vs(0.085 卤1.11 卤0.31 卤1.43 卤0.05 卤1.67 卤0.49)vs(0.64 卤0.10], and the ability of colony formation and migration were significantly decreased (P 0.05). Compared with the control group, the expression of Tyr and Trp1 increased significantly in the P0.05MAPK signaling pathway, and the expression of p-P38 and p-JNK increased significantly in the P0.05MAPK signaling pathway, but there was no significant difference in the activity of p-ERK between the two groups. Conclusion by upregulating the activities of p-P38 and p-JNK in the MAPK signaling pathway, the cell proliferation of mouse melanoma B16 cells was inhibited obviously, and the differentiation of melanoma cells was induced by cell differentiation in morphology and function.
【作者单位】：河北医科大学第四医院肿瘤研究所&科研中心;华北制药集团新药研究开发有限责任公司;
【基金】：河北省卫生厅医学科学研究重点课题项目基金资助(No.20120120)~~
【分类号】：R739.5

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