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[1]张姣惠 张 颖 林心宇 黄小红**.姜黄素调控p53蛋白入核诱导骨髓瘤细胞凋亡*[J].应用与环境生物学报,2019,25(04):1-8.[doi:10.19675/j.cnki.1006-687x.2018.11046]
 ZHANG Jiaohui,ZHANG Ying,LIN Xinyu & HUANG Xiaohong**.Curcumin regulates P53 protein entry into nucleus to induce apoptosis of P3X63Ag8*[J].Chinese Journal of Applied & Environmental Biology,2019,25(04):1-8.[doi:10.19675/j.cnki.1006-687x.2018.11046]
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姜黄素调控p53蛋白入核诱导骨髓瘤细胞凋亡*()
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《应用与环境生物学报》[ISSN:1006-687X/CN:51-1482/Q]

卷:
25卷
期数:
2019年04期
页码:
1-8
栏目:
研究论文
出版日期:
2019-07-30

文章信息/Info

Title:
Curcumin regulates P53 protein entry into nucleus to induce apoptosis of P3X63Ag8*
文章编号:
201811046
作者:
张姣惠 张 颖 林心宇 黄小红**
福建农林大学动物科学学院 福州 350002
Author(s):
ZHANG Jiaohui ZHANG Ying LIN Xinyu & HUANG Xiaohong**
College of Animal Sciences, Fujian Agriculture and Forestry University, Fuzhou 350002, China
关键词:
姜黄素骨髓瘤细胞p53凋亡
Keywords:
curcumin P3X63Ag8 cells p53 apoptosis
DOI:
10.19675/j.cnki.1006-687x.2018.11046
摘要:
探讨姜黄素对小鼠骨髓瘤细胞核内p53蛋白含量的影响及对其凋亡作用。体外培养P3X63Ag8细胞,给予不同浓度姜黄素处理(0、10、20、30、40、50 ?mol/L)24 h,通过还原性辅酶Ⅰ氧化法测定LDH活性、Caspase 3 荧光分析试剂盒检测Caspase 3 活性的变化、采用2’,7’-二氯荧光黄双乙酸盐(DCFH-DA)法检测细胞活性氧(ROS)量的变化,利用Western-Blot技术检测细胞质与细胞核中p53蛋白分布情况。结果显示,姜黄素浓度为20 ?mol/L时,P3X63Ag8细胞LDH活性极显著升高(P < 0.01),并在40 ?mol/L组达到顶峰。30 ?mol/L姜黄素处理后,Caspase-3活性出现极显著的提高(P < 0.01),其余姜黄素处理组与未处理组相比没有呈现显著变化的现象。与阴性对照组相比,当姜黄素浓度为30 ?mol/L时细胞内活性氧水平显著上升(P < 0.01),而随着姜黄素浓度提高到40和50 ?mol/L,细胞内活性氧水平均开始极显著下降(P < 0.01),且高浓度组下降程度高于中浓度组。从细胞质和细胞核蛋白分离的试验结果可以看出,在细胞质蛋白中,p53蛋白含量随着姜黄素浓度的增加而降低,与此相反,细胞核蛋白中p53蛋白含量则呈现了上升趋势。因此,姜黄素能调控LDH活性、Caspase-3活性以及活性氧水平来促进细胞损伤,从而抑制骨髓瘤细胞P3X63Ag8的增殖,促进其凋亡。在蛋白水平上促进p53蛋白入核,从而进一步刺激细胞凋亡程序的启动。
Abstract:
This study is aimed to investigate the effect of curcumin on the protein level of p53 in the nucleus of mouse myeloma cells and its effect of apoptosis. P3X63Ag8 cells were cultured with different concentrations of curcumin (0, 10, 20, 30, 40, 50 ?mol/L) for 24 h. LDH activity was determined by reducing coenzyme I oxidation method. Caspase 3 was detected by caspase 3 fluorescence assay kit. 2’,7’-dichlorofluorescein diacetate (DCFH-DA) method was used to detect changes in cellular reactive oxygen species (ROS), and the cytoplasmic proteins and nuclear protein were separated to dectect the protein level of P53 in cytoplasm and nucleus by western-blot. The results showed that when the concentration of curcumin was 20 μmol/L, the activity of LDH in P3X63Ag8 cells was significantly increased (P < 0.01), and reached the peak under 40 μmol/L. Treatment with 30 μmol/L curcumin, the activity of Caspase-3 was significantly increased (P < 0.01), and the other curcumin treatment groups showed no significant difference compared with the control group. Compared with the negative control group, the level of reactive oxygen species was significantly increased when the concentration of curcumin was 30 μmol/L (P < 0.01), and the level of reactive oxygen species showed a significant decrease (P < 0.01), when the concentration of curcumin was 40 and 50 μmol/L. From the western blot results, it can be seen that in the cytoplasmic protein, the protein level of p53 was decreased with the increased concentration of curcumin. In contrast, the protein level of p53 in nuclear showed an upward trend. To sum up, curcumin can regulate the activity of LDH, caspase-3 and ROS to promote cell damage and thereby inhibiting the proliferation of myeloma cells P3X63Ag8 and promoting its apoptosis. In addition, Promoting p53 protein entry into the nucleus and regulate apoptosis-related proteins at the protein level.

备注/Memo

备注/Memo:
收稿日期: 2018-11-26 接受日期 Accepted: 2019-01-04
*国家自然科学基金项目(31572484)资助?
**通讯作者(E-mail: xhhuang138@hotmail.com)
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更新日期/Last Update: 2019-01-24