[1]杨崇,傅若秋,高宁.8-姜酚抑制乳腺癌细胞增殖的分子机制研究[J].第三军医大学学报,2019,41(05):406-414.
 YANG Chong,FU Ruoqiu,GAO Ning.Molecular mechanism of inhibition of 8-gingerol on proliferation of breast cancer cells[J].J Third Mil Med Univ,2019,41(05):406-414.
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8-姜酚抑制乳腺癌细胞增殖的分子机制研究(/HTML )
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《第三军医大学学报》[ISSN:1000-5404/CN:51-1095/R]

卷:
41卷
期数:
2019年第05期
页码:
406-414
栏目:
基础医学
出版日期:
2019-03-15

文章信息/Info

Title:
Molecular mechanism of inhibition of 8-gingerol on proliferation of breast cancer cells
作者:
杨崇傅若秋高宁
陆军军医大学(第三军医大学)药学与检验医学系生药学教研室
Author(s):
YANG Chong FU Ruoqiu GAO Ning

Department of Pharmacognosy, Faculty of Pharmacy & Laboratory Medicine, Army Medical University (Third Military Medical University), Chongqing, 400038, China

关键词:
8-姜酚乳腺癌活性氧P38周期阻滞
Keywords:
8-gingerol breast cancer reactive oxygen species P38 cell cycle arrest
分类号:
R285.5; R73-36; R737.9
文献标志码:
A
摘要:

目的 探讨8-姜酚抑制乳腺癌细胞增殖的分子机制。方法 MTT法测定乳腺癌细胞MDA-MB-231细胞活性;流式细胞仪检测产生活性氧(reactive oxygen species,ROS)的细胞比例、细胞周期以及细胞凋亡;Western blot检测周期相关蛋白Cyclin D1和p-DCD2的表达。结果 8-姜酚能明显抑制乳腺癌MDA-MB-231和MCF-7细胞的增殖活性,差异有统计学意义(P<0.01),72 h的IC50值分别为(22.06±2.93)、(35.77±5.96)μmol/L;流式细胞仪检测结果发现8-姜酚可导致乳腺癌细胞G1期阻滞,诱导细胞产生ROS;8-姜酚作用于MDA-MB-231和MCF-7细胞24 h后G1期细胞的百分比分别为(69.23±3.77)%、(65.87±3.76)%,与对照组G1期的细胞比例(47.39±1.97)%、(49.17±3.52)%相比,差异具有统计学意义(P<0.01);用NAC抑制ROS后减弱了8-姜酚对乳腺癌细胞G1期的阻滞作用,用P38抑制剂抑制P38蛋白磷酸化水平后也减弱了8-姜酚对乳腺癌细胞G1期的阻滞作用。Western blot结果显示8-姜酚可以上调MAPK信号通路中P38蛋白的磷酸化水平;并可下调G1期相关蛋白Cyclin D1和p-CDC2的表达,用NAC抑制ROS后减弱了8-姜酚对P38磷酸化水平上调作用的同时还减弱了8-姜酚对周期相关蛋白CyclinD1、p-CDC2的下调作用;用P38抑制剂抑制P38蛋白磷酸化水平后同样减弱了8-姜酚对周期相关蛋白CyclinD1、p-CDC2的下调作用。 结论 8-姜酚具有明显的抑制肿瘤增殖活性的作用,其作用机制可能为8-姜酚通过诱导乳腺癌细胞产生ROS从而导致MAPK信号通路中P38的磷酸化激活,进而引起乳腺癌细胞的周期阻滞。

Abstract:

Objective To investigate the effect of cell cycle on breast cancer cells by 8-gingerol. Methods The cell viability of breast cancer cells MDA-MB-231 and MCF-7 after treated by 8-gingerol at the doses of 0, 10, 20, 30 or 40 μmol/L for 24, 48 or 72 h was assessed by MTT assay. The cell cycle, apoptosis and proportion of cells producing ROS were measured by flow cytometry. The expression of cell cycle associated proteins, Cyclin D1 and p-CDC2 were detected by Western blot analysis. Results 8-gingerol evidently inhibited the proliferation of breast cancer MDA-MB-231 and MCF-7 cells (P<0.01). The inhibitory concentration of IC50 were 22.06±2.93 and 35.77±5.96 μmol/L, respectively in the breast cancer MDA-MB-231 and MCF-7 cells after 72 h treatment with 8-gingerol. The results of flow cytometry showed that 8-gingerol can cause G1 phase arrest and induce cells to produce ROS in the breast cancer MDA-MB-231 and MCF-7 cells after treatment with 8-gingerol for 72 h. Flow cytometry found that 8-gingerol resulted in cell cycle arrest at G1 phase cells was (69.23±3.77)% and (65.87±3.76)%, respectively in the MDA-MB-231 and MCF-7 cells after 8-gingerol treatment for 24 h, significantly higher than in the cells without treatment [(47.39±1.97)%,(49.17±3.52)%, P<0.01]. Pretreatment with NAC weakened the effect of 8-gingerol on G1 phase arrest. Western blot results showed that 8-gingerol resulted in the decreases of Cyclin D1 and p-CDC2 levels, and upregulated the phosphorylation level of P38. The inhibition of ROS by NAC attenuated the up-regulation of phosphorylation of P38 by 8-gingerol and the down-regulation of cyclin-related proteins Cyclin D1 and p-CDC2. The inhibition of P38 phosphorylation by P38 inhibitors also attenuated the down-regulation of cyclin-associated proteins Cyclin D1 and p-CDC2 by 8-gingerol. Conclusion 8-gingerol exhibits obviously inhibitory effects on breast cancer cell proliferation. The mechanism may be that 8-gingerol induces ROS in breast cancer cells, which leads to phosphorylation of P38 in the MAPK signaling pathway, and then leads to cell cycle arrest.

 

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更新日期/Last Update: 2019-03-05