PO.ET09.05 · 实验与分子治疗
DNA损伤、凋亡和细胞周期紊乱：一种新型锡(IV)配合物的抗肿瘤效应
DNA damage, apoptosis and cell cycle disruption: Antitumor effects of a novel tin(IV) complexes
作者与单位 Authors & Affiliations
摘要 Abstract
中文摘要
引言：以顺铂为代表的铂类化疗药物被广泛用于抗癌，但受限于毒性和耐药性。有机锡(IV)配合物提供了一种有前景的替代方案，具有增强的疗效和独特的作用机制。在此，我们评估了二丁基和二苯基有机锡(IV)配合物的抗癌活性，并研究了它们在多种癌细胞系中的作用机制。
材料与方法：有机锡(IV)配合物的合成：二丁基和二苯基有机锡(IV)（Bu-Sn和Ph-Sn）配合物按照既定方案合成。使用光谱技术验证配合物的纯度和结构确认。抗癌活性：本研究使用了以下人类细胞系：T-47D（乳腺癌）、HCT116（结直肠癌）、AsPC-1（胰腺腺癌）和MCF-10A（非致瘤性乳腺上皮）。按照厂商方案，利用MTT法测定半数最大抑制浓度（IC50）。通过流式细胞术使用Annexin V和碘化丙啶（PI）染色法研究细胞系中的细胞死亡方式（凋亡和坏死）。以顺铂作为参比药物，通过流式细胞术进行细胞周期分析，以确定每个细胞系细胞周期的变化。通过ICP-MS评估有机锡(IV)配合物的亚细胞分布。将处理后的细胞分级分离为主要细胞区室，以确定配合物的主要定位。使用彗星实验评估有机锡(IV)配合物诱导的DNA损伤，从而定量与处理相关的DNA链断裂。
结果与讨论：Bu-Sn在所有测试的癌细胞系中均表现出强效细胞毒性活性（IC50<3.00 μM），对T-47D人乳腺癌细胞的效力尤其高（IC50=0.53 μM）。重要的是，它对非癌性MCF-10细胞显示出极小的毒性，表现出良好水平的选择性。相比之下，Ph-Sn仅显示出有限的细胞毒性活性。从机制上讲，Bu-Sn诱导T-47D细胞G1期阻滞并触发剂量依赖性凋亡。亚细胞分析显示以细胞核蓄积为主（Bu-Sn为32%，Ph-Sn为24%），这与彗星实验中观察到的显著DNA损伤相关，提示细胞核靶向和基因毒性应激是其抗癌活性的基础。
结论：合成并全面表征了两种含二丁基和二苯基基团的有机锡(IV)配合物。二丁基配合物在多种癌细胞系中表现出强效细胞毒性活性，同时不损伤非恶性细胞。机制研究表明，这些锡配合物主要靶向DNA，诱导细胞周期阻滞和凋亡，最终导致癌细胞死亡。
查看英文原文 English abstract
Introduction: Platinum-based chemotherapeutics like cisplatin are widely used against cancer but are limited by toxicity and drug resistance. Organotin(IV) complexes offer a promising alternative with enhanced efficacy and distinct mechanisms. Here, we evaluated the anticancer activity of dibutyl and diphenyl organotin(IV) complexes and investigated their mechanisms of action across multiple cancer cell lines.
Material and Method: Synthesis of Organotin(IV) Complexes: Dibutyl and diphenyl organotin(IV) (Bu-Sn and Ph-Sn) complexes were synthesized following established protocols. The purity and structural confirmation of the complexes were verified using spectroscopic techniques. Anti-cancer activities: The following human cell lines were used in this study: T-47D (breast carcinoma), HCT116 (colorectal carcinoma), AsPC-1 (pancreatic adenocarcinoma), and MCF-10A (non-tumorigenic mammary epithelial). Following manufactures protocol MTT assay is utilized for the determination of half maximum inhibitory concentration (IC 50 ). The mode of cell death (apoptosis and necrosis) investigated in cell lines via flow cytometry using the Annexin V and propidium iodide (PI) staining method. Analysis of cell cycle with flow cytometry performed to determine the changes in the cell cycle for each cell lines utilizing cisplatin as a reference drug. Subcellular distribution of the organotin(IV) complexes was evaluated by ICP-MS. Treated cells were fractionated into major cellular compartments to determine the predominant localization of the complexes. DNA damage induced by the organotin(IV) complexes was assessed using the Comet assay, allowing quantification of treatment-related DNA strand breaks.
Results and Discussion: Bu-Sn exhibited strong cytotoxic activity across all tested cancer cell lines (IC₅₀ < 3.00 µM), with particularly high potency against T-47D human breast cancer cells (IC₅₀ = 0.53 µM). Importantly, it showed minimal toxicity toward non-cancerous MCF-10 cells, demonstrating a favorable level of selectivity. In contrast, Ph-Sn displayed only limited cytotoxic activity. Mechanistically, Bu-Sn induced G1 phase arrest and triggered dose-dependent apoptosis in T-47D cells. Subcellular analysis revealed predominant nuclear accumulation (32% for Bu-Sn, 24% for Ph-Sn), correlating with pronounced DNA damage observed in Comet assays, suggesting that nuclear targeting and genotoxic stress underlie its anticancer activity.
Conclusion: Two organotin(IV) complexes containing dibutyl and diphenyl groups were synthesized and fully characterized. The dibutyl complex exhibited strong cytotoxic activity across multiple cancer cell lines while sparing non-malignant cells. Mechanistic studies indicated that these tin complexes primarily target DNA, inducing cell cycle arrest and apoptosis, ultimately leading to cancer cell death.
利益披露 Disclosure
S. Sharma, None..
K. Damodaran, None.