蛹虫草素和黄酮与启动子 DNA 序列 d(CACGTG)2 的结合研究：多光谱研究和细胞毒性评估

IF 4 2区化学 Q2 CHEMISTRY, PHYSICAL

Journal of Molecular Structure Pub Date : 2024-11-16 DOI:10.1016/j.molstruc.2024.140765

Shailendra Kumar, Chandrachur Ghosh, Partha Roy, Maya S. Nair

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引用次数: 0

摘要

利用多种光谱技术探讨了菊黄素和黄酮与 d(CACGTG)2 DNA 序列的相互作用。吸收光谱和荧光光谱结果表明，蛹虫草素与黄酮存在部分插层作用，而黄酮则存在外部作用。黄菊素和黄酮的结合系数（Kb）值分别为 105 和 104 M-1。利用荧光数据绘制的工作图证实，菊黄素 DNA 复合物的化学计量为 1:1，而黄酮 DNA 复合物在溶液中有多种化学计量。荧光寿命分析数据显示，与 DNA 复合物结合后，蛹素的平均寿命延长，而黄酮的平均寿命缩短。用 EtBr 和 Hoechst 33258 进行的竞争性置换研究数据表明存在部分插层。圆二色性研究结果表明，在与菊黄素和黄酮结合后，DNA 结构发生了变化。圆二色性和差示扫描量热法的热数据显示，在 1:1 的复合物中，蛹素-DNA 的熔化温度发生了微小变化，而黄酮 DNA 复合物则没有显著变化。这些熔化温度的微小变化支持了菊黄素和黄酮的非交联结合。游离的 d(CACGTG)2 DNA 与黄菊素和黄酮相互作用后，其热焓变化显著增加。具体来说，对于菊粉，ΔH1 从 88.07 千卡/摩尔增加到 196.40 千卡/摩尔，ΔH2 从 41.14 千卡/摩尔增加到 104.00 千卡/摩尔。就黄酮而言，ΔH1 从 88.07 千卡/摩尔上升到 127.40 千卡/摩尔，ΔH2 从 41.14 千卡/摩尔上升到 180.70 千卡/摩尔。范德华力和疏水相互作用是 d(CACGTG)2 DNA 与蛹素和黄酮结合的关键因素。利用 MTT 试验和 AO/EtBr 染色法分析了菊黄素和黄酮的细胞毒性作用。MTT数据显示，它们对MCF-7、DU-145和HCT-15癌细胞株有明显的抗增殖作用，其中黄酮对DU-145前列腺癌细胞最有效，而菊黄素对MCF-7乳腺癌细胞的作用更大。菊黄素和黄酮对 MCF-7、HCT-15 和 DU-145 的 IC50 值分别为 74.97 µM、95.15 µM、88.66 µM、78.62 µM，以及 149.45 µM、45.17 µM。与对照细胞相比，黄酮化合物在所有三种癌细胞系中都能大幅诱导细胞凋亡。这项研究确认黄酮类化合物是潜在的促凋亡剂。

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Binding investigation of chrysin and flavone with promoter DNA sequence d(CACGTG)2: Multi spectroscopic studies and cytotoxic evaluation

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Binding investigation of chrysin and flavone with promoter DNA sequence d(CACGTG)2: Multi spectroscopic studies and cytotoxic evaluation

Interactions of chrysin and flavone with d(CACGTG)₂ DNA sequence were explored using multi spectroscopic techniques. Absorption and fluorescence spectroscopic results indicated the partial intercalation for chrysin, and flavone interact externally. The binding coefficient (K_b) values of chrysin and flavone are 10⁵ and 10⁴ M^-1, respectively. Job plots using fluorescence data confirmed the 1:1 stoichiometry for the chrysin DNA complex, while multiple stoichiometries for the flavone DNA complex in solutions. Fluorescence lifetime analysis data showed that the average lifetime of chrysin increased while the average lifetime of flavone decreased upon complexation with DNA. Competitive displacement studies with EtBr and Hoechst 33258 data suggest partial intercalation. Circular dichroism results showed that DNA structures were perturbed upon chrysin and flavone binding. Circular dichroism and differential scanning calorimetry thermal data showed minute changes in melting temperature at 1:1 complex for chrysin-DNA, while there was no significant change in flavone DNA complex. These slight changes in melting temperatures support the non-intercalative binding of chrysin and flavone. The heat enthalpy changes of the free d(CACGTG)₂ DNA increased significantly upon interaction with both chrysin and flavone. Specifically, for chrysin, ΔH₁ increased from 88.07 to 196.40 kcal/mol and ΔH₂ from 41.14 to 104.00 kcal/mol. In the case of flavone, ΔH₁ rose from 88.07 to 127.40 kcal/mol and ΔH₂ from 41.14 to 180.70 kcal/mol. The van der Waals forces and hydrophobic interactions are key contributors to the binding between d(CACGTG)₂ DNA and both chrysin and flavone. The cytotoxic effect of chrysin and flavone was analyzed using an MTT assay and AO/EtBr staining. MTT data showed that they exhibited a significant antiproliferation effect against MCF-7, DU-145, and HCT-15 cancer cell lines, where flavone is most effective against DU-145 prostate cancer cells, and chrysin has more effects against MCF-7 breast cancer cells. The IC₅₀ values of chrysin and flavone are 74.97 µM, 95.15 µM, 88.66 µM, 78.62 µM and 149.45 µM, 45.17 µM against MCF-7, HCT-15, and DU-145 respectively. Flavone compounds induced apoptosis substantially in all three cancer cell lines compared to the control cells. This study recognizes flavones as potential proapoptotic agents.

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来源期刊

Journal of Molecular Structure 化学-物理化学

CiteScore

7.10

自引率

15.80%

发文量

2384

审稿时长

45 days

期刊介绍： The Journal of Molecular Structure is dedicated to the publication of full-length articles and review papers, providing important new structural information on all types of chemical species including: • Stable and unstable molecules in all types of environments (vapour, molecular beam, liquid, solution, liquid crystal, solid state, matrix-isolated, surface-absorbed etc.) • Chemical intermediates • Molecules in excited states • Biological molecules • Polymers. The methods used may include any combination of spectroscopic and non-spectroscopic techniques, for example: • Infrared spectroscopy (mid, far, near) • Raman spectroscopy and non-linear Raman methods (CARS, etc.) • Electronic absorption spectroscopy • Optical rotatory dispersion and circular dichroism • Fluorescence and phosphorescence techniques • Electron spectroscopies (PES, XPS), EXAFS, etc. • Microwave spectroscopy • Electron diffraction • NMR and ESR spectroscopies • Mössbauer spectroscopy • X-ray crystallography • Charge Density Analyses • Computational Studies (supplementing experimental methods) We encourage publications combining theoretical and experimental approaches. The structural insights gained by the studies should be correlated with the properties, activity and/ or reactivity of the molecule under investigation and the relevance of this molecule and its implications should be discussed.