| 抗菌肽Dermaseptin-PP协同化疗药物抗肿瘤并逆转A549/DDP细胞顺铂耐药性 |
| 投稿时间：2024-04-05 点此下载全文
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| 引用本文：闫丽文,胡祖成,彭凤栖,胡文均,胡海燕,陈基杰,陆洋.抗菌肽Dermaseptin-PP协同化疗药物抗肿瘤并逆转A549/DDP细胞顺铂耐药性[J].中国现代中药,2024,26(10):1727-1738 |
| DOI：10.13313/j.issn.1673-4890.20240405002 |
| 摘要点击次数: 366 |
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| 作者中文名 | 作者英文名 | 单位中文名 | 单位英文名 | E-Mail |
| 闫丽文 |
YAN Li-wen |
北京中医药大学 中药学院，北京 102488 |
School of Chinese Materia Medica, Beijing University of Chinese Medicine, Beijing 102488, China |
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| 胡祖成 |
HU Zu-cheng |
北京中医药大学 中药学院，北京 102488 |
School of Chinese Materia Medica, Beijing University of Chinese Medicine, Beijing 102488, China |
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| 彭凤栖 |
PENG Feng-qi |
北京中医药大学 中药学院，北京 102488 |
School of Chinese Materia Medica, Beijing University of Chinese Medicine, Beijing 102488, China |
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| 胡文均 |
HU Wen-jun |
北京中医药大学 中药学院，北京 102488 |
School of Chinese Materia Medica, Beijing University of Chinese Medicine, Beijing 102488, China |
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| 胡海燕 |
HU Hai-yan |
北京中医药大学 中药学院，北京 102488 |
School of Chinese Materia Medica, Beijing University of Chinese Medicine, Beijing 102488, China |
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| 陈基杰 |
JAKKREE Tangthianchaichana |
泰国国立法政大学 朱拉蓬国际医学院，巴吞他尼府 12120 |
Chulabhorn International College of Medicine, Thammasat University, Pathum Thani 12120, Thailand |
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| 陆洋* |
LU Yang |
北京中医药大学 中药学院，北京 102488 |
School of Chinese Materia Medica, Beijing University of Chinese Medicine, Beijing 102488, China |
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| 基金项目:国家中医药管理局高水平中医药重点学科建设项目（zyyzdxk-2023272） |
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| 中文摘要:目的 探究抗菌肽Dermaseptin-PP对多种化疗药物体外抗肿瘤活性的协同增效作用，以及其对人非小细胞肺癌顺铂（DDP）耐药细胞株A549/DDP耐药性的逆转作用，并进一步探究其作用机制。方法 采用噻唑蓝（MTT）染色法检测不同质量浓度的Dermaseptin-PP对人非小细胞肺癌A549、H157细胞或小鼠乳腺癌4T1细胞的增殖抑制作用，考察低毒质量浓度Dermaseptin-PP与多烯紫杉醇（DTX）、阿霉素（DOX）或DDP联用对A549、H157或4T1细胞的协同抗肿瘤作用；采用细胞计数试剂盒-8（CCK-8）法检测低毒质量浓度Dermaseptin-PP对A549/DDP细胞DDP耐药性的逆转作用。采用流式细胞术检测Dermaseptin-PP对DDP诱导A549细胞凋亡的影响，采用乳酸脱氢酶（LDH）释放实验及碘化丙啶（PI）摄取实验考察Dermaseptin-PP对A549和A549/DDP细胞膜完整性的影响，最后通过扫描电镜观察Dermaseptin-PP处理后A549和A549/DDP细胞膜形态的变化。结果 Dermaseptin-PP可在体外抑制A549、H157和4T1细胞增殖且呈剂量依赖性；与低毒质量浓度Dermaseptin-PP（0.002、0.020、0.200 μg·mL–1）联用后，DTX、DOX及DDP对A549细胞的增殖抑制率均有不同程度的提高，半数抑制浓度（IC50）均有所降低，药物联合指数（CI）基本均小于0.9，其中Dermaseptin-PP与DDP的协同作用最强（CI<0.4）。此外，Dermaseptin-PP与DDP联用对H157和4T1细胞的CI也均小于0.9，表现为协同作用。低毒质量浓度Dermaseptin-PP（0.2、2.0、4.0 μg·mL–1）均能显著逆转A549/DDP细胞的DDP耐药性，逆转倍数（RF）分别为2.83、5.09、9.73。相比于单用DDP，与0.2 μg·mL–1 Dermaseptin-PP联用可显著提高A549细胞的凋亡率（P<0.05）；不同质量浓度的Dermaseptin-PP均可显著增加A549、A549/DDP细胞的LDH释放率和PI摄取率，且呈浓度依赖性；扫描电镜观察发现，经2×IC50的Dermaseptin-PP处理后，A549细胞的细胞膜明显受损、质膜解体、细胞形态发生改变，A549/DDP细胞的细胞膜也出现明显孔洞。结论 Dermaseptin-PP对多种化疗药物体外抗肿瘤活性均具有协同增效作用，其中与DDP的协同作用最强，并且能够逆转A549/DDP细胞的DDP耐药性，其增效及逆转耐药的机制可能与破坏肿瘤细胞膜，进而提高化疗药物入胞药量有关。 |
| 中文关键词:抗菌肽 Dermaseptin-PP 化疗 抗肿瘤 耐药 顺铂 |
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| Dermaseptin-PP Combined with Chemotherapeutic Agents Treats Tumors and Reverses Cisplatin Resistance of A549/DDP Cells |
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| Abstract:Objective To investigate the effects of the antimicrobial peptide Dermaseptin-PP combined with chemotherapeutic agents on the tumor cells in vitro and the cisplatin (DDP) resistance of A549/DDP cells, and further explore the underlying mechanisms.Methods The methyl thiazolyl tetrazolium (MTT) assay was employed to determine the inhibitory effects of different concentrations of Dermaseptin-PP on the proliferation of A549, H157 or 4T1 cells and the inhibitory effects of low concentrations of Dermaseptin-PP in combination with docetaxel (DTX), doxorubicin (DOX) or DDP on A549, H157 or 4T1 cells. The cell-counting kit 8 (CCK-8) assay was used to determine the reversal effects of low concentrations of Dermaseptin-PP on the DDP resistance of A549/DDP cells. Flow cytometry was employed to measure the effect of Dermaseptin-PP on DDP-induced apoptosis of A549 cells. The lactate dehydrogenase (LDH) release assay and propidium iodide (PI) uptake assay were employed to investigate the effect of Dermaseptin-PP on the membrane integrity of A549 and A549/DDP cells. Finally, the morphological changes of A549 and A549/DDP cell membranes after Dermaseptin-PP treatment were observed by scanning electron microscopy.Results Dermaseptin-PP inhibited the proliferation of A549, H157, and 4T1 cells in vitro in a dose-dependent manner. In combination with low concentrations (0.002, 0.020, and 0.200 μg·mL–1) of Dermaseptin-PP, DTX, DOX, and DDP demonstrated increased inhibition rates on the proliferation of A549 cells, with reduced half maximal inhibitory concentrations (IC50) and the combination indexes (CIs) lower than 0.9. The combination of Dermaseptin-PP with DDP demonstrated the strongest synergistic effect (CI<0.4). In addition, Dermaseptin-PP combined with DDP showed a synergistic effect, with the CIs lower than 0.9 for both H157 and 4T1 cells. Low concentrations (0.2, 2.0, and 4.0 μg·mL–1) of Dermaseptin-PP all significantly reversed the DDP resistance of A549/DDP cells, with the reversal folds (RFs) of 2.83, 5.09, and 9.73, respectively. Compared with DDP alone, the combination of DDP with 0.2 μg·mL–1 Dermaseptin-PP increased the apoptosis rate of A549 cells (P<0.05). Dermaseptin-PP increased the LDH release rate and PI uptake rate of A549 and A549/DDP cells in a concentration-dependent manner. The treatment with Dermaseptin-PP at a concentration of 2×IC50 damaged the cell membranes, disintegrated the plasma membranes, and altered the morphology of A549 cells, and the treatment led to the appearance of holes in the cell membranes of A549/DDP cells.Conclusion Dermaseptin-PP combined with chemotherapeutic agents demonstrated synergistic effects on the tumor cells in vitro, among which the synergistic effect with DDP was the strongest. Moreover, Dermaseptin-PP can reverse the DDP resistance of A549/DDP cells by directly destroying the cell membrane to improve the amount of chemotherapeutic agents entering the cells. |
| keywords:antimicrobial peptides Dermaseptin-PP chemotherapy anti-tumor drug resistance cisplatin (DDP) |
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