Gao Yu, Yang Zhanyi, Bajpai Akhilesh Kumar, Wang Wenben, Zhang Liyuan, Xia Zhenhong
Department of Pharmacy, Binzhou Medical University, Yantai, China.
Department of Genetics, Genomics and Informatics, University of Tennessee Health Science Center, Memphis, TN, United States.
Front Oncol. 2024 Sep 24;14:1453164. doi: 10.3389/fonc.2024.1453164. eCollection 2024.
In this study, we aimed to explore the mechanism by which resveratrol promotes cisplatin-induced death of HepG2 cells and to provide a potential strategy for resveratrol in the treatment of cancer.
HepG2 cells were exposed to a range of drug concentrations for 24 h: resveratrol (2.5 μg/mL [10.95 μM], 5 μg/mL [21.91 μM], 10 μg/mL [43.81 μM], 20 μg/mL [87.62 μM], 40 μg/mL [175.25 μM], and 80 μg/mL [350.50 μM]), cisplatin (0.625 μg/mL [2.08 μM], 1.25 μg/mL [4.17 μM], 2.5 μg/mL [8.33 μM], 4.5 μg/mL [15.00 μM], and 10 μg/mL [33.33 μM]), 24 μg/mL (105.15 μM) resveratrol + 9 μg/mL (30.00 μM) cisplatin, and 12 μg/mL (52.57 μM) resveratrol + 4.5 μg/mL (15.00 μM) cisplatin. The interaction of two drugs was evaluated by coefficient of drug interaction (CDI), which was based on the Pharmacological Additivity model. The MTT 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay was used to detect the effect of different concentrations of drugs on cell viability, while transcriptome sequencing was used to identify pathways associated with higher gene enrichment. Synchrotron radiation FTIR microspectroscopy experiments and data analysis were conducted to obtain detailed spectral information. The second-derivative spectra were calculated using the Savitzky-Golay algorithm. Single-cell infrared spectral absorption matrices were constructed to analyze the spectral characteristics of individual cells. The Euclidean distance between cells was calculated to assess their spectral similarity. The cell-to-cell Euclidean distance was computed to evaluate the spatial relationships between cells. The target protein of resveratrol was verified by performing a Western blot analysis.
After 24 h of treatment with resveratrol, HepG2 cell growth was inhibited in a dose-dependent manner. Resveratrol promotes cisplatin-induced HepG2 cell death through membrane-related pathways. It also significantly changes the membrane components of HepG2 cells. Additionally, resveratrol changes the morphology of the HepG2 cell membrane by decreasing the expression of PLA2G2.
Resveratrol changes the morphology of the HepG2 cell membrane by decreasing the expression of PLA2G2 and promotes cisplatin-induced HepG2 cell death. The combination of cisplatin and resveratrol can play a synergistic therapeutic effect on HepG2 cells.
在本研究中，我们旨在探究白藜芦醇促进顺铂诱导的HepG2细胞死亡的机制，并为白藜芦醇在癌症治疗中提供潜在策略。
将HepG2细胞暴露于一系列药物浓度下24小时：白藜芦醇（2.5μg/mL [10.95μM]、5μg/mL [21.91μM]、10μg/mL [43.81μM]、20μg/mL [87.62μM]、40μg/mL [175.25μM]和80μg/mL [350.50μM]）、顺铂（0.625μg/mL [2.08μM]、1.25μg/mL [4.17μM]、2.5μg/mL [8.33μM]、4.5μg/mL [15.00μM]和10μg/mL [33.33μM]）、24μg/mL（105.15μM）白藜芦醇+9μg/mL（30.00μM）顺铂以及12μg/mL（52.57μM）白藜芦醇+4.5μg/mL（15.00μM）顺铂。基于药理相加模型，通过药物相互作用系数（CDI）评估两种药物的相互作用。采用MTT 3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐法检测不同浓度药物对细胞活力的影响，同时利用转录组测序鉴定与基因富集程度较高相关的通路。进行同步辐射傅里叶变换红外光谱显微实验及数据分析以获取详细光谱信息。使用Savitzky-Golay算法计算二阶导数光谱。构建单细胞红外光谱吸收矩阵以分析单个细胞的光谱特征。计算细胞间的欧几里得距离以评估其光谱相似性。计算细胞到细胞的欧几里得距离以评估细胞间的空间关系。通过蛋白质免疫印迹分析验证白藜芦醇的靶蛋白。
用白藜芦醇处理24小时后，HepG2细胞生长受到剂量依赖性抑制。白藜芦醇通过膜相关途径促进顺铂诱导的HepG2细胞死亡。它还显著改变HepG2细胞的膜成分。此外，白藜芦醇通过降低PLA2G2的表达改变HepG2细胞膜的形态。
白藜芦醇通过降低PLA2G2的表达改变HepG2细胞膜的形态，并促进顺铂诱导的HepG2细胞死亡。顺铂与白藜芦醇联合应用可对HepG2细胞发挥协同治疗作用。
