归芪白术方有效小分子芦荟大黄素调控MDM2/p53信号通路促进胃癌细胞凋亡和周期阻滞的机制研究

王雯; 苏韫; 张晗; 龚红霞; 曹旺杰; 王芳; 黄勇; 刘永琦

doi:10.13748/j.cnki.issn1007-7693.20251727

归芪白术方有效小分子芦荟大黄素调控MDM2/p53信号通路促进胃癌细胞凋亡和周期阻滞的机制研究

Mechanism of the Active Small Molecule Aloe-emodin from Guiqi Baizhu Formula in Inducing Gastric Cancer Cell Apoptosis and Cycle Arrest via the MDM2/p53 Signaling Pathway

摘要

摘要:
目的探讨归芪白术方有效小分子芦荟大黄素调控MDM2/p53信号通路促进胃癌细胞凋亡和周期阻滞的机制。
方法通过分子对接技术分析芦荟大黄素与MDM2蛋白的结合活性；运用CCK-8法检测胃癌细胞增殖能力；利用划痕试验检测细胞迁移能力；借助流式细胞术分析细胞周期及凋亡比例；通过Western blotting检测MDM2/p53信号通路相关蛋白MDM2、p53、Cyclin D1、c-Myc、Bcl-2、Bax的表达水平；通过RT-qPCR技术检测MDM2/p53信号通路中MDM2、p53、Cyclin D1、c-Myc、Bcl-2、Bax mRNA的表达水平。
结果分子对接结果显示，与川陈皮素和槲皮素相比，芦荟大黄素与MDM2具有较强结合能力；CCK-8法试验表明，与0 μmol·L⁻¹组相比，芦荟大黄素能以浓度和时间依赖性的方式抑制HGC-27、AGS细胞增殖(P<0.05或P<0.01)；流式试验结果表明，与0 μmol·L⁻¹组相比，芦荟大黄素可使HGC-27细胞G1期比例呈浓度依赖性升高，且在12.5 μmol·L⁻¹和25 μmol·L⁻¹时作用显著(P<0.01)，AGS细胞G1期比例也呈浓度依赖性升高(P<0.01)；划痕试验显示，与0 μmol·L⁻¹组相比，12.5 μmol·L⁻¹和25 μmol·L⁻¹的芦荟大黄素可抑制HGC-27、AGS细胞迁移能力；Western blotting和qRT-PCR试验结果表明，与0 μmol·L⁻¹组相比，不同浓度芦荟大黄素干预HGC-27、AGS细胞后，能下调MDM2、CyclinD1、c-Myc、Bcl-2蛋白和基因的表达，并且上调p53和Bax蛋白和基因的表达(P<0.05或P<0.01)。
结论芦荟大黄素可以通过靶向MDM2/p53信号通路，抑制胃癌细胞增殖、诱导凋亡和周期阻滞。

Abstract:
OBJECTIVE To investigate how aloe-emodin, an active compound derived from the Guiqi Baizhu Formula, promotes apoptosis and cell cycle arrest in gastric cancer cells by regulating the MDM2/p53 signaling pathway.
METHODS Molecular docking technology was utilized to characterize the binding activity between aloe-emodin and the MDM2 protein. CCK-8 assays were used to detect the proliferation ability of gastric cancer cells, and the evaluation of migratory capacity through wound healing assays. Flow cytometric analysis was conducted to quantify cell cycle distribution profiles and apoptosis rates. Western blotting was used to determine protein expression levels of key MDM2/p53 signaling pathway components(MDM2, p53, Cyclin D1, c-Myc, Bcl-2, and Bax), complemented by RT-qPCR analysis of corresponding mRNA expression patterns for these targets.
RESULTS Molecular docking demonstrated substantial binding affinity between aloe-emodin and MDM2. CCK-8 assays indicated concentration- and time-dependent inhibition of HGC-27 and AGS cell proliferation(P<0.05 or P<0.01). Flow cytometry confirmed that aloe-emodin significantly increased the proportion of HGC-27 and AGS cells in G1 phase (P<0.01). Wound healing assays showed concentration-dependent suppression of cell migration at 12.5 μmol·L⁻¹ and 25 μmol·L⁻¹ concentrations. Both Western blotting and qRT-PCR analyses consistently revealed that aloe-emodin treatment downregulated MDM2, Cyclin D1, c-Myc, and Bcl-2 expression while upregulating p53 and Bax at protein and mRNA levels across tested concentrations(P<0.05 or P<0.01).
CONCLUSION Aloe-emodin from Guiqi Baizhu Formula demonstrates anti-tumor effects in gastric cancer by targeting the MDM2/p53 pathway, effectively inhibiting cell proliferation, inducing apoptosis, and triggering cell cycle arrest.

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