Citrate-capped AuNPs-induced redox reprogramming modulates the TP53–BAX/BCL2–CASP3 Axis, reinforcing antioxidant defense and promoting apoptotic signaling in liver Cancer

IF 4.6 2区化学 Q1 SPECTROSCOPY

Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy Pub Date : 2025-07-01 DOI:10.1016/j.saa.2025.126630

Alaa Elmetwalli , Sara Abdelsayed , Mervat G. Hassan , Ibtisam Aboud Almutairi , Deema Kamal Sabir , Hassnaa Elsherbiny , Ashraf Elsayed

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Abstract

Background

Hepatocellular carcinoma (HCC) remains a leading cause of cancer-related mortality worldwide, characterized by limited therapeutic options and poor prognosis. Nanomedicine offers promising approaches to overcome conventional treatment limitations through enhanced targeting capabilities. Gold nanoparticles (AuNPs) have garnered attention due to their exceptional physicochemical properties and biocompatibility. This study investigated the selective anti-cancer activity of citrate-capped AuNPs against HepG2 liver cancer cells compared to normal human fibroblasts (WI-38), focusing on molecular mechanisms underlying differential cytotoxicity.

Methods

Citrate-capped AuNPs were synthesized using a modified citrate reduction technique and characterized through complementary analytical methods (FTIR, TEM, XRD, zeta potential, UV–vis spectroscopy). Cytotoxicity was assessed via MTT assay and phase-contrast microscopy. Gene expression analysis using qRT-PCR examined key apoptosis-related markers (TP53, BAX, BCL2, CASP3). Mitochondrial membrane potential was evaluated using Rhodamine-123 fluorescence. Oxidative stress parameters and antioxidant enzyme activities were comprehensively analyzed to elucidate redox modulation.

Results

Synthesized AuNPs exhibited spherical morphology (32.6 nm), high crystallinity, and strong negative surface charge (−35 mV). AuNPs demonstrated selective cytotoxicity toward HepG2 cells (IC₅₀: 28.48 μg/mL) compared to WI-38 cells (IC₅₀: 464 μg/mL). Treatment upregulated TP53 and BCL2 expression while downregulating BAX and CASP3, creating a complex apoptotic signaling pattern. AuNPs induced significant mitochondrial membrane potential collapse (82 % reduction at 50 μg/mL). Remarkably, treatment enhanced antioxidant enzyme activities (SOD: +30 %, CAT: +22.4 %, GPx: +15 %) while reducing oxidative stress markers (ROS: -38.3 %, MDA: −30.3 %), suggesting redox reprogramming rather than oxidative damage.

Conclusion

Citrate-capped AuNPs selectively induce cytotoxicity in HepG2 cells through TP53 activation, mitochondrial dysfunction, and redox reprogramming. Despite upregulated antioxidant defenses, cancer cells ultimately succumb to AuNPs-induced mitochondrial damage. The differential sensitivity between cancer and normal cells suggests a promising therapeutic potential, leveraging cancer cells' unique metabolic vulnerabilities while minimizing damage to healthy tissues.

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柠檬酸盐覆盖的aunps诱导的氧化还原重编程调节TP53-BAX / BCL2-CASP3轴，增强抗氧化防御并促进肝癌细胞凋亡信号传导

肝细胞癌（HCC）仍然是世界范围内癌症相关死亡的主要原因，其特点是治疗选择有限，预后差。纳米医学提供了有希望的方法，通过增强靶向能力来克服传统治疗的局限性。金纳米颗粒（AuNPs）由于其特殊的物理化学性质和生物相容性而引起了人们的关注。本研究研究了柠檬酸盐封顶的AuNPs与正常人成纤维细胞（WI-38）相比，对HepG2肝癌细胞的选择性抗癌活性，重点研究了差异细胞毒性的分子机制。方法采用改进的柠檬酸还原技术合成柠檬酸盐包盖AuNPs，并采用互补分析方法（FTIR、TEM、XRD、zeta电位、UV-vis光谱）对其进行表征。采用MTT法和相衬显微镜检测细胞毒性。基因表达分析采用qRT-PCR检测关键凋亡相关标志物（TP53， BAX, BCL2, CASP3）。罗丹明-123荧光法测定线粒体膜电位。综合分析氧化应激参数和抗氧化酶活性来阐明氧化还原调控。结果合成的AuNPs呈球形（32.6 nm），结晶度高，表面带强负电荷（- 35 mV）。与WI-38细胞（IC₅₀：464 μg/mL）相比，AuNPs对HepG2细胞表现出选择性的细胞毒性（IC₅₀：28.48 μg/mL）。治疗上调了TP53和BCL2的表达，同时下调了BAX和CASP3的表达，形成了复杂的凋亡信号模式。AuNPs显著诱导线粒体膜电位塌陷（50 μg/mL时降低82%）。值得注意的是，处理提高了抗氧化酶活性（SOD: + 30%, CAT: + 22.4%, GPx: + 15%），同时降低了氧化应激标志物（ROS: - 38.3%, MDA: - 30.3%），表明氧化还原重编程而不是氧化损伤。结论柠檬酸盐封顶的AuNPs通过TP53激活、线粒体功能障碍和氧化还原重编程选择性诱导HepG2细胞毒性。尽管抗氧化防御上调，癌细胞最终还是屈服于aunps诱导的线粒体损伤。癌细胞和正常细胞之间的敏感性差异表明，利用癌细胞独特的代谢脆弱性，同时最大限度地减少对健康组织的损害，这是一种有希望的治疗潜力。

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来源期刊

Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy 物理-光谱学

CiteScore

8.40

自引率

11.40%

发文量

1364

审稿时长

40 days

期刊介绍： Spectrochimica Acta, Part A: Molecular and Biomolecular Spectroscopy (SAA) is an interdisciplinary journal which spans from basic to applied aspects of optical spectroscopy in chemistry, medicine, biology, and materials science. The journal publishes original scientific papers that feature high-quality spectroscopic data and analysis. From the broad range of optical spectroscopies, the emphasis is on electronic, vibrational or rotational spectra of molecules, rather than on spectroscopy based on magnetic moments. Criteria for publication in SAA are novelty, uniqueness, and outstanding quality. Routine applications of spectroscopic techniques and computational methods are not appropriate. Topics of particular interest of Spectrochimica Acta Part A include, but are not limited to: Spectroscopy and dynamics of bioanalytical, biomedical, environmental, and atmospheric sciences, Novel experimental techniques or instrumentation for molecular spectroscopy, Novel theoretical and computational methods, Novel applications in photochemistry and photobiology, Novel interpretational approaches as well as advances in data analysis based on electronic or vibrational spectroscopy.