Yin-Hua Yuan, Qi Guang, Shan-Shan Zhang, Mi Tang
Cancer Research Center, Zhumadian Central Hospital of Henan Province Zhumadian 463000, China.
West China School of Public Health, Sichuan University Chengdu 610039, China.
Zhongguo Zhong Yao Za Zhi. 2020 Nov;45(22):5495-5503. doi: 10.19540/j.cnki.cjcmm.20200819.303.
Orthogonal experiments were used to optimize the process parameters of curcumin TPP-PEG-PCL nanomicelles; the particle size, electric potential and morphology under the electron microscope were systematically detected for the curcumin TPP-PEG-PCL nanomicelles; and the stability and in vitro release of the curcumin TPP-PEG-PCL nanomicelles were investigated. With DID fluorescent dye as the fluorescent probe, flow cytometry was used to study the uptake of nanomicelles by breast cancer cells, and laser confocal microscopy was used to study the mitochondrial targeting and lysosomal escape functions of nanomicelles. Under the same dosage conditions, the effect of curcumin TPP-PEG-PCL nanomicelles on promoting the apoptosis of breast cancer cells was evaluated. The optimal particle size of curcumin TPP-PEG-PCL nanomicelle was(17.3±0.3) nm, and the Zeta potential was(14.6±2.6) mV in orthogonal test. Under such conditions, the micelle appeared as regular spheres under the transmission electron microscope. Fluorescence test results showed that TPP-PEG-PCL nanomicelles can promote drug uptake by tumor cells, escape from lysosomal phagocytosis, and target the mitochondria. The cell survival rate and Hoechst staining positive test results showed that curcumin TPP-PEG-PCL nanomicelles had a good effect on promoting apoptosis of breast cancer cells. The curcumin TPP-PEG-PCL micelles can significantly reduce the mitochondrial membrane potential of breast cancer cells, increase the release of cytochrome C, significantly increase the expression of pro-apoptotic protein Bcl-2 and reduce the expression of anti-apoptotic Bax protein. These test results were significantly better than those of curcumin PEG-PCL nanomicelles and curcumin, with statistically significant differences. The results revealed that curcumin TPP-PEG-PCL nanomicelles can well target breast cancer cell mitochondria and escape from the lysosomal capture, thereby enhancing the drug's role in promoting tumor cell apoptosis.
采用正交试验优化姜黄素TPP-PEG-PCL纳米胶束的工艺参数；系统检测姜黄素TPP-PEG-PCL纳米胶束的粒径、电势及电子显微镜下的形态；研究姜黄素TPP-PEG-PCL纳米胶束的稳定性及体外释放情况。以DID荧光染料为荧光探针，采用流式细胞术研究纳米胶束被乳腺癌细胞摄取的情况，并用激光共聚焦显微镜研究纳米胶束的线粒体靶向和溶酶体逃逸功能。在相同给药剂量条件下，评价姜黄素TPP-PEG-PCL纳米胶束对促进乳腺癌细胞凋亡的作用。正交试验中姜黄素TPP-PEG-PCL纳米胶束的最佳粒径为(17.3±0.3) nm，Zeta电位为(14.6±2.6) mV。在此条件下，胶束在透射电子显微镜下呈规则球形。荧光检测结果表明，TPP-PEG-PCL纳米胶束可促进肿瘤细胞摄取药物，从溶酶体吞噬中逃逸，并靶向线粒体。细胞存活率和Hoechst染色阳性检测结果表明，姜黄素TPP-PEG-PCL纳米胶束对促进乳腺癌细胞凋亡有良好作用。姜黄素TPP-PEG-PCL胶束可显著降低乳腺癌细胞的线粒体膜电位，增加细胞色素C的释放，显著增加促凋亡蛋白Bcl-2的表达并降低抗凋亡蛋白Bax的表达。这些检测结果明显优于姜黄素PEG-PCL纳米胶束和姜黄素，差异具有统计学意义。结果表明，姜黄素TPP-PEG-PCL纳米胶束可很好地靶向乳腺癌细胞线粒体并从溶酶体捕获中逃逸，从而增强药物促进肿瘤细胞凋亡的作用。
