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| 过氧化氢响应的嵌段共聚物纳米胶束对乳腺癌MCF-7细胞的杀伤效应 |
| The killing effect of hydrogen peroxide-responsive block copolymeric nanomicelles on breast cancer MCF-7 cell line |
| 投稿时间：2024-08-28 |
| DOI：10.3969/j.issn.1000-0399.2025.05.001 |
| 中文关键词: 可控聚合|阿霉素|响应性缓释|乳腺癌细胞系|谷胱甘肽 |
| 英文关键词: Controllable polymerization|Doxorubicin|Responsive release|Breast cancer cell line|Glutathione |
| 基金项目:国家自然科学基金面上项目(编号:31870993) |
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| 摘要点击次数: 143 |
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| 中文摘要: |
| 目的 探究负载阿霉素（DOX）的聚合物纳米胶束DOX@PBEM对乳腺癌MCF-7细胞的细胞毒性效果。方法 可控聚合方法合成了一种具有亲水段和疏水段的两亲性嵌段共聚物（PBEM），通过自组装溶剂挥发法制备了包载DOX的、过氧化氢（H2O2）响应的聚合物纳米胶束DOX@PBEM，并对聚合物性质和胶束体外药物释放行为开展研究。利用激光共聚焦显微镜观察MCF-7细胞在0、1、2、4小时的胶束摄取特征，采用荧光显微镜观察MCF-7细胞内谷胱甘肽（GSH）、活性氧（ROS）水平，采用噻唑蓝比色（MTT）和细胞死活染色法研究PBEM和DOX@PBEM对MCF-7细胞的生长抑制作用。结果 本研究成功制备（77.6±6.4）nm的DOX@PBEM，并在RPMI-1640培养基（含10%胎牛血清）中稳定保存。在PBS（pH 7.4，含100μmol H2O2）中孵育2 d,DOX@PBEM实现了63.5%DOX的释放，孵育3 d后，尺寸从（81.5±4.8）nm增加到（227.4±3.5）nm。激光共聚焦显微成像（CLSM）结果 显示，DOX@PBEM能被MCF-7细胞有效摄取。相较于不做处理组，50μmol/L有效单元的纳米胶束处理后，MCF-7细胞胞内GSH荧光信号由PBS处理组（41.9±1.2）降低至（19.7±0.6），胞内ROS荧光信号由PBS处理组（6.2±0.6）增加至（15.5±1.6）。MTT结果显示，PBEM聚合物对MCF-7细胞没有毒性，而当DOX浓度为10μg/m L时，DOX@PBEM处理的MCF-7细胞仅有（30.0±1.8）%的细胞存活率；细胞死活染色结果佐证DOX@PBEM能达到游离DOX相似的细胞毒性效果。结论 构建的DOX@PBEM能有效地被乳腺癌细胞摄取，并且在细胞内微环境下响应性缓慢释放DOX和醌甲基小分子，放大细胞内氧化还原微环境的失衡，实现对乳腺癌细胞的高效杀伤作用。 |
| 英文摘要: |
| Objective To investigate the cytotoxic effect of doxorubicin （DOX）-loaded polymeric nanomicelles （DOX@PBEM） on breast cancer MCF-7 cells. Methods An amphiphilic block copolymer （PBEM） with hydrophilic and hydrophobic segments was synthesized by controlled polymerization.Hydrogen peroxide（H2O2）-responsive polymeric nanomicelle loaded with DOX（DOX@PBEM） was prepared by self-assembly solvent evaporation method.Then properties of polymers and drug release behavior of the micelle in vitro were studied.The cell uptake characteristics of nanomicelle for MCF-7 cells at 0, 1, 2 and 4 h were observed by confocal laser scanning microscope （CLSM）.The levels of Glutathione （GSH） and Hydrogen peroxide （H2O2） in MCF cells were measured by Fluorescence microscope.Lastly, MTT assay and livedead cell staining were applied to assess the inhibitory effects of PBEM and DOX@PBEM on MCF-7 cell line. Results DOX@PBEM with（77.6±6.4） nm was successfully prepared.The prepared micelle kept stable in RPMI-1640 medium （containing 10% FBS）.After incubation in PBS （p H 7.4, containing 100μmol H2O2） for two days, DOX@PBEM released 63.5%DOX, and the size increased from （81.5±4.8） nm to（227.4±3.5） nm after incubation for three days.CLSM results showed that DOX@PBEM could be efficiently taken up by MCF-7 cells.Furthermore, compared with the untreated group, the intracellular GSH fluorescence signal of MCF-7 cells decreased from （41.9±1.2） to （19.7±0.6）and the intracellular ROS fluorescence signal increased from （6.2±0.6） to （15.5±1.6） after the treatment with 50μM effective unit nanoparticles.MTT analysis showed that PBEM polymer was not toxic to MCF-7 cells, but only （30.8±1.8）%of MCF-7 cells kept alive after treated with DOX@PBEM when the concentration of DOX was 10μg/m L.The results of live-dead cell staining also proved that the cytotoxicity of DOX@PBEM was similar to that of free DOX. Conclusion The constructed DOX@PBEM can be effectively accumulated into breast cancer cells, then DOX and quinone methyl release responsively in the intracellular microenvironment under stimulation of ROS, enlarging the imbalance of intracellular redox microenvironment and achieving high-efficiency killing effect on breast cancer cells. |
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