[1]赵宁,夏利敏,宋涛.藏红花素通过PI3K/Akt/GSK-3β通路诱导人乳腺癌细胞凋亡的初步研究[J].西部中医药,2024,37(04):17-22.[doi:10.12174/j.issn.2096-9600.2024.04.04]
　ZHAO Ning,XIA Limin,SONG Tao.Preliminary Study of Saffronin-induced Apoptosis of Human Breast Cancer Cells via PI3K/Akt/GSK-3β Pathway[J].Western Journal of Traditional Chinese Medicine,2024,37(04):17-22.[doi:10.12174/j.issn.2096-9600.2024.04.04]
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藏红花素通过PI3K/Akt/GSK-3β通路诱导人乳腺癌细胞凋亡的初步研究
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《西部中医药》[ISSN:2096-9600/CN:62-1204/R]

卷:
37
期数:
2024年04期
页码:
17-22
栏目:
基础研究
出版日期:
2024-04-15

文章信息/Info

Title:
Preliminary Study of Saffronin-induced Apoptosis of Human Breast Cancer Cells via PI3K/Akt/GSK-3β Pathway
作者:
赵宁, 夏利敏, 宋涛
邯郸市中心医院，河北 邯郸 056001
Author(s):
ZHAO Ning, XIA Limin, SONG Tao
Handan Central Hospital, Handan 056001, China
关键词:
乳腺癌; 藏红花素; 增殖; 细胞凋亡; PI3K/Akt/GSK-3信号通路
Keywords:
breast cancer; saffronin; proliferation; cellular apoptosis; p-PI3K/Akt/GSK-3signaling pathway
分类号:
R737.9
DOI:
10.12174/j.issn.2096-9600.2024.04.04
文献标志码:
A
摘要:
目的探讨藏红花素对人乳腺癌细胞凋亡的影响及相关调控机制。 方法以人正常乳腺MCF-10A细胞和人乳腺癌MCF-7细胞为受试细胞，分别以不同浓度藏红花素0（空白对照组）、200、400、800 mg/L和LY294002（PI3K特异性抑制剂）15 mg/L干预对数生长期MCF-10A细胞和MCF-7细胞48 h，采用四甲基偶氮唑蓝（MTT）法、克隆形成实验分别检测各组细胞增殖抑制率、细胞克隆能力，Annexin V-FITC染色法检测细胞凋亡水平，运用蛋白免疫印迹法（Western blot）检测磷脂酰肌醇3-激酶（phosphatidylinositol 3-kinase，PI3K）、磷酸化PI3K（p-PI3K）、蛋白激酶B（protein kinase B，Akt）、磷酸化Akt（p-Akt）、糖原合成酶激酶-3β（glycogen synthase kinase-3β，GSK-3β）、磷酸化GSK-3β（p-GSK-3β）、半胱氨酸天门冬氨酸蛋白酶9（cysteine aspartic proteases-9，Caspase-9）、激活型半胱氨酸天门冬氨酸蛋白酶3（cleaved cysteine aspartate protease-3，Cleaved caspase-3）、B淋巴细胞瘤2（B-cell lymphoma-2，Bcl-2）、Bcl-2相关X蛋白（bcl-2 related X protein，Bax）蛋白表达。 结果与空白对照组比较，200、400、800 mg/L藏红花素组和LY294002组MCF-10A细胞增殖抑制率、克隆数目、凋亡率，差异均无统计学意义（P＞0.05）；400、800 mg/L藏红花素组和LY294002组MCF-7细胞增殖抑制率升高、克隆数目降低、凋亡率升高（P＜0.01）；p-PI3K、p-Akt、p-GSK-3β、Bcl-2表达下调且Caspase-9、Cleaved Caspase-3、Bax表达上调（P＜0.01），磷酸化率p-PI3K/PI3K、p-Akt/Akt、p-GSK-3β/GSK-3β降低（P＜0.01），Bax/Bcl-2表达比值升高（P＜0.01）。与LY294002组比较，800 mg/L藏红花素组MCF-7细胞增殖抑制率升高、克隆数目降低、凋亡率升高（P＜0.05或P＜0.01）；Caspase-9、Cleaved Caspase-3、Bax表达上调（P＜0.05或P＜0.01），Bax/Bcl-2比值升高（P＜0.01），其他指标两组间比较差异无统计学意义（P＞0.05）。 结论藏红花素具有诱导人乳腺癌细胞凋亡的作用，其机制可能与抑制PI3K/Akt/GSK-3β信号通路有关。
Abstract:
ObjectiveTo explore the effect of saffronin on the apoptosis of human breast cancer cells and related regulatory mechanisms. MethodsHuman normal breast MCF-10A cells and human breast cancer MCF-7 cells were used as test cells, and different concentrations of saffronin 0 (blank control group), 200, 400, 800 mg/L and LY294002 (PI3K-specific inhibitor) 15 mg/L were used to intervene in the logarithmic growth phase MCF-10A cells and MCF-7 cells for 48 h. Tetramethyl azole blue (MTT) assay and clonal formation assay were adopted to detect the inhibition rate of cell proliferation and clonogenic ability of cells in each group respectively, Annexin V-FITC staining to detect the level of apoptosis, Western blot was applied to detect the expressions of PI3K, p-PI3K, Akt, p-Akt, GSK-3β, p-GSK-3β, Caspase-9, Cleaved caspase-3, Bcl-2 and Bax. ResultsCompared with the blank control group, the difference had no statistical meaning in the proliferation inhibition rate, clone number and apoptosis rate of MCF-10A cells in the 200, 400 and 800 mg/L saffronin group and the LY294002 group (P>0.05); the proliferation inhibition rate of MCF-7 cells in 400, 800 mg/L saffronin group and LY294002 group increased, the number of clones decreased, and the apoptosis rate increased (P<0.01); the expression of p-PI3K, p-Akt, p-GSK-3β, Bcl-2 was down-regulated, and the expression of Caspase-9, Cleaved Caspase-3, and Bax was up-regulated (P<0.01). The phosphorylation rates of p-PI3K/PI3K, p-Akt/Akt, p-GSK-3β /GSK-3β were decreased (P<0.01), and the Bax/Bcl-2 expression ratio was increased (P<0.01). Compared with the LY294002 group, MCF-7 cells in the 800 mg/L saffronin group showed higher inhibition of cell proliferation, lower number of clones, and higher apoptosis rate (P<0.05 or P<0.01); Caspase-9, Cleaved Caspase-3 and Bax expressions were up-regulated (P<0.05 or P<0.01), and the Bax/Bcl-2 ratio was elevated (P<0.01), while the differences of the other indexes were not statistically significant between the two groups (P>0.05). Conclusionsaffronin could induce the apoptosis of human breast cancer cells, and its mechanism might be related to inhibiting p-PI3K/Akt/GSK-3β signaling pathway.

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备注/Memo

备注/Memo:
赵宁（1981—），女，主治医师。研究方向：乳腺疾病的诊治及研究。E-mail：zhn8101@163.com。河北省医学科学研究课题计划（20191643）。
更新日期/Last Update: 2024-04-15