黑磷纳米片通过线粒体途径介导三阴性乳腺癌细胞凋亡的研究  

Black phosphorus nanosheets induce apoptosis of triple-negative breast cancer cells mediated by mitochondrial apoptosis pathway

作　　者：赵旭 马晓雅 吴国举[2] 马洁 袁伟 Zhao Xu;Ma Xiaoya;Wu Guoju;Ma Jie;Yuan Wei(State Key Laboratory of Molecular Oncology,National Cancer Center,National Clinical Research Center for Cancer,Cancer Hospital,Chinese Academy of Medical Sciences and Peking Union Medical College,Beijing 100021,China;Department of General Surgery,Beijing Hospital,National Center of Gerontology,Institute of Geriatric Medicine,Chinese Academy of Medical Sciences,Beijing 100730,China;Center of Biotherapy,Beijing Hospital,National Center of Gerontology,Institute of Geriatric Medicine,Chinese Academy of Medical Sciences,Beijing 100730,China)

机构地区：[1]国家癌症中心、国家肿瘤临床医学研究中心、中国医学科学院北京协和医学院肿瘤医院、分子肿瘤学国家重点实验室,北京100021 [2]北京医院、国家老年医学中心、中国医学科学院老年医学研究院普外科,北京100730 [3]北京医院、国家老年医学中心中国、医学科学院老年医学研究院生物治疗中心,北京100730

出　　处：《中国医学前沿杂志（电子版）》2023年第4期25-32,共8页Chinese Journal of the Frontiers of Medical Science(Electronic Version)

基　　金：国家自然科学基金面上项目(51972343);中央高水平医院临床科研业务费资助(BJ-2022-196);中央高校基本科研业务费专项资金资助(E2ET5104X2)。

摘　　要：目的探讨聚乙二醇(polyethylene glycol,PEG)修饰的黑磷纳米片(black phosphorus nanosheets,BPNS)对三阴性乳腺癌(triple-negative breast cancer,TNBC)MDA-MB-231和4T1细胞凋亡的影响及生物学机制。方法通过液相剥离方法制备BPNS,再经PEG修饰获得BP-PEG。利用透射电子显微镜、动态光散射对BPNS和BP-PEG进行表征。通过紫外-可见吸收光谱及透射电子显微镜评估两者在超纯水中的稳定性。利用乳酸脱氢酶细胞毒性法,检测BP-PEG对鼠源和人源的正常和肿瘤细胞活力的影响;采用γ-H2AX免疫荧光染色检测肿瘤细胞DNA损伤的情况;通过细胞流式术检测细胞凋亡情况;利用JC-1染色检验肿瘤细胞线粒体膜电位变化情况;Western blot检测BP-PEG对Cyt c、Bcl-2和Bax等线粒体凋亡相关蛋白表达水平的影响。结果BPNS为层状纳米结构,PEG修饰后平均直径和电位略有增加,并且可有效提高BPNS的稳定。BP-PEG浓度为50μg/ml范围内表现出对肿瘤细胞良好的杀伤活性。BP-PEG可造成TNBC细胞DNA损伤并促进细胞凋亡,引起线粒体膜电位Δψm下降,可显著上调Cyt c和Bax并降低Bcl2的表达水平。结论BPPEG可通过线粒体凋亡途径有效促进TNBC细胞凋亡,并对正常细胞表现出较好的应用安全性,其可能成为一个潜在的肿瘤治疗药物。Objective To investigate the effects of polyethylene glycol(PEG)modified black phosphorus nanosheets(BPNS)on the apoptosis of MDA-MB-231 and 4T1 cells of triple-negative breast cancer and its biological mechanism.Methods BPNS were prepared by liquid-phase exfoliation,then modified with PEG.BPNS and BP-PEG were characterized by transmission electron microscopy(TEM)and dynamic light scattering(DLS).The stability of BPNS and BP-PEG in ultrapure water were evaluated via UV-visible absorption spectroscopy and TEM.Lactate dehydrogenase cytotoxicity assay was used to test the effect of BP-PEG on normal and tumor cell viability.DNA damage of tumor cells were detected byγ-H2AX immunofluorescence staining.The apoptosis ability was measured by flow cytometry.The changes of mitochondrial membrane potential of tumor cells were detected by JC-1 staining.Western blot analysis was performed to detect the expression levels of cytochrome c,Bcl-2 and Bax after BP-PEG treatment.Results The BPNS were layered nanostructures.The average diameter and potential of BPNS were slightly increased after PEG modification,and the stability could be effectively improved.BP-PEG exhibited tumor-killing activity within the concentration range of 50μg/ml.BP-PEG may cause DNA damage and promote apoptosis of triple-negative breast cancer cells,and decrease mitochondrial membrane potential.Western blot showed that the expression level of cytochrome c and Bax were increased,and expression of Bcl2 was decreased.Conclusions BP-PEG could effectively promote apoptosis of triple-negative breast cancer cells through mitochondrial apoptosis pathway,and showed better application safety to normal cells,which may become a potential anti-tumor agent.

关 键 词：黑磷纳米片 三阴性乳腺癌 线粒体凋亡途径 

分 类 号：R737.9[医药卫生—肿瘤]