School of Life Science and Institute for Advanced Materials, South China Normal University, Guangzhou 510631, China.
Biomaterials. 2012 Sep;33(26):6162-71. doi: 10.1016/j.biomaterials.2012.05.037. Epub 2012 Jun 8.
The aim of this study is to reveal the biological mechanism for high anti-cancer efficiency of co-immobilized TNF-α plus IFN-γ polymeric drug (co-immobilized drug) in mediating two gynecologic cancer cell lines: MCF-7 and OVCAR-3. The co-immobilized drug is prepared by mixing 10 ng/ml TNF-α plus 10 ng/ml IFN-γ which are then photo-immobilized onto cell culture polystyrene plates. The drug compositions and microstructures are characterized by Fourier transform infrared spectroscopy and scanning electron microscopy. The MCF-7 and OVCAR-3 cell cycle arrest and programmed cell death are checked by flow cytometry, and the expression of p53 is probed by immunofluorescence staining. The phosphorylation sites of the p53 regulation and the apoptosis key protein expressions of caspase 3, 8 and 9 are detected by western blot assay. Our data show that, in case of short treatment time (48 h) at low cytokine concentrations (20 ng/ml), the co-immobilized drug demonstrates visible effects in comparison with the treatment using TNF-α plus IFN-γ freely attached on the polymeric plate (free drug). It is revealed that the co-immobilized drug leads to significant cell arrest in the S phase or G(1) and G(2) phase and offer high efficiency in mediating a caspase-dependent apoptosis via p53 transcriptional regulation. Moreover, upon the treatment by the co-immobilized drug, the two gynecologic cancer cell lines show different phosphorylation sites of p53 and then different caspase-dependent apoptosis pathways. The present work sheds deep insights into the p53 regulation mechanism responsible for the high anti-cancer efficiency of the co-immobilized TNF-α plus IFN-γ polymeric drug against MCF-7 and OVCAR-3.
本研究旨在揭示共固定 TNF-α 加 IFN-γ 聚合物药物（共固定药物）介导两种妇科癌细胞系 MCF-7 和 OVCAR-3 高抗癌效率的生物学机制。共固定药物是通过将 10ng/ml TNF-α 加 10ng/ml IFN-γ 混合，然后光固定在细胞培养聚苯乙烯板上制备的。药物成分和微观结构通过傅里叶变换红外光谱和扫描电子显微镜进行表征。通过流式细胞术检查 MCF-7 和 OVCAR-3 细胞周期停滞和程序性细胞死亡，并通过免疫荧光染色探测 p53 的表达。通过 Western blot 检测 p53 调节的磷酸化位点和凋亡关键蛋白 caspase 3、8 和 9 的表达。我们的数据表明，在低细胞因子浓度（20ng/ml）和短处理时间（48h）的情况下，与自由附着在聚合物板上的 TNF-α 加 IFN-γ （游离药物）相比，共固定药物表现出明显的效果。结果表明，共固定药物导致 S 期或 G1 和 G2 期的细胞明显停滞，并通过 p53 转录调节介导高效的 caspase 依赖性凋亡。此外，在用共固定药物处理后，两种妇科癌细胞系显示出不同的 p53 磷酸化位点，然后是不同的 caspase 依赖性凋亡途径。本工作深入了解了共固定 TNF-α 加 IFN-γ 聚合物药物对 MCF-7 和 OVCAR-3 高抗癌效率负责的 p53 调节机制。
