| 刘玲琳,毛娟娟,岑梦燕,杨扬.从自噬和副凋亡途径初探桔梗皂苷-D抗乳腺癌作用与机理[J].浙江中西医结合杂志,2024,34(8): |
| 从自噬和副凋亡途径初探桔梗皂苷-D抗乳腺癌作用与机理 |
| Preliminary study on the anti-tumor effect and mechanism of platycodon-D through autophagy and paraapoptosis pathway |
| 投稿时间：2023-03-05 修订日期：2024-07-22 |
| DOI： |
| 中文关键词: 桔梗皂苷-D 乳腺肿瘤 自噬 副凋亡 胞质空泡化 |
| 英文关键词:Platycodin-D breast cacinoma autophagy paraptosis cell vacuolization |
| 基金项目:宁波市自然科学基金（202003N4213）；宁波市中医医学重点学科建设计划项(2022-Z04) |
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| 中文摘要: |
| 目的：从自噬和副凋亡途径探讨中医传统引经药“桔梗”有效成分桔梗皂苷-D（PD）抗乳腺癌的作用和潜在机理。方法：以乳腺癌细胞株4T1、MDA-MB-231、MCF-7作为体外研究对象，结晶紫法检测PD对三种细胞存活率的影响；MDC荧光标记法与 LC3B荧光染色激光共聚焦显微镜观察PD作用乳腺癌细胞后自噬发生的情况；通过自噬抑制后明确PD诱导自噬在乳腺癌细胞死亡中的作用；光镜与电镜下观察PD作用乳腺癌细胞后形态学变化的影响。结果：PD 作用4T1、M231细胞后存活率明显下降，当浓度分别为10 μM、20 μM、40 μM时均具有统计学差异。MDC法与 LC3B荧光染色发现经PD处理的三种乳腺癌细胞后发生自噬现象，而PD联合自噬抑制剂氯喹的加入反而显著降低了三种细胞的存活率（具统计学差异）。光镜与电镜观察经PD处理的三种细胞内出现大量空泡结构，并伴有线粒体肿胀，类似副凋亡的形态学改变。结论：PD具有明确的抑制乳腺癌细胞4T1、MDA-MB-231、MCF-7存活率的效应，该效应伴随自噬现象的发生，但自噬抑制剂的加入提示PD诱导的细胞自噬是对自身的保护作用。经PD处理的三种乳腺癌细胞出现类似副凋亡的形态学改变，初步推测PD对乳腺癌细胞的抑制作用可能与副凋亡相关。 |
| 英文摘要: |
| ABSTRACT Objective To confirm the anti-tumor activity and potential mechanism of Platycodin D through autophagy and paraapoptotic pathway. Methods Breast cancer cell lines 4T1, M231 and MCF-7 were used as in vitro study tools to determine the inhibition rate influenced by PD through crystal violet method. The effect of PD on the autophagy was evaluated by MDC fluorescent and laser confocal microscopy. The viability rate of 3 breast cancer line cells were compared with PD vs PD combined with chloroquine.Morphological changes of PD on 4T1, M231 and MCF-7 cells by microscope and electron microscope. Results PD can significantly reduce the survival rate of 4T1, M231, MCF-7 cells, and when the PD’s concentration is 10mM、20mM and 40mM, the inhibition rate of cells is elevated, which have a definite quantitative relationship.In the MDC assay, PD causes cell autophagy. PD can induce the expression of intracellular LC3B?by laser confocal microscopy.But under the condition of blocking autophagy by chloroquine , PD significantly reduced the survival rate of three breast cancer cells.By light microscopy and electron microscopy, the vacuolization phenomenon was found influenced by PD. Conclusion PD has a definite anti-tumor effect on breast cancer cell lines 4T1, MDA-MB-231 and MCF-7,which is accompanied by the occurrence of autophagy, but the effect is proved not autophagy dependencies.Morphology changes similar to paraapoptosis are observed from the light microscopy and electron microscopy，which are different from the typical apoptosis.We speculate that the potential mechanism of inhibitory effect of PD on breast cancer cells may be related to paraapoptosis. |
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