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丝裂霉素C电化学疗法通过抑制鳞状癌细胞的自噬增强凋亡性死亡。

Electrochemotherapy with Mitomycin C Potentiates Apoptosis Death by Inhibiting Autophagy in Squamous Carcinoma Cells.

作者信息

Condello Maria, D'Avack Gloria, Vona Rosa, Spugnini Enrico Pierluigi, Scacco Licia, Meschini Stefania

机构信息

National Center for Drug Research and Evaluation, National Institute of Health, 00161 Rome, Italy.

Center for Gender-Specific Medicine, National Institute of Health, 00161 Rome, Italy.

出版信息

Cancers (Basel). 2021 Jul 31;13(15):3867. doi: 10.3390/cancers13153867.

DOI:10.3390/cancers13153867
PMID:34359775
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC8345561/
Abstract

We investigated the chemosensitizing effect of electroporation (EP), which, using electrical pulses, permeabilizes cancer cells to drugs. The study involved two human hypopharyngeal and tongue carcinoma cell lines. The surface and intracytoplasmic expression of P-gp were evaluated by flow cytometry, demonstrating that both lines were intrinsically resistant. After establishing the optimal dose of mitomycin C (MMC) to be used, in combination with EP, we showed, by both MTT assay and optical and electron scanning microscopy, the potentiating cytotoxic effect of EP with MMC compared to single treatments. Flow cytometry showed that the cytotoxicity of EP + MMC was due to the induction of apoptosis. In addition to verifying the release of cytochrome C in EP + MMC samples, we performed an expression analysis of caspase-3, caspase-9, Akt, pAkt, HMGB1, LC3I, LC3II, p62, Beclin1, and associated proteins with both apoptotic and autophagic phenomena. Our results were confirmed by two veterinary patients in whom the EP + MMC combination was used to control margins after the resection of corneal squamous carcinoma. In conclusion, we affirmed that the effect for which EP enhances MMC treatment is due to the inhibition of the autophagic process induced by the drug in favor of apoptosis.

摘要

我们研究了电穿孔（EP）的化学增敏作用，电穿孔利用电脉冲使癌细胞对药物通透。该研究涉及两个人下咽和舌癌细胞系。通过流式细胞术评估P-糖蛋白（P-gp）的表面和胞浆内表达，结果表明这两个细胞系均具有内在抗性。在确定了与EP联合使用的丝裂霉素C（MMC）的最佳剂量后，我们通过MTT法以及光学和电子扫描显微镜显示，与单一治疗相比，EP与MMC联合使用具有增强的细胞毒性作用。流式细胞术表明，EP + MMC的细胞毒性是由于诱导了细胞凋亡。除了验证EP + MMC样品中细胞色素C的释放外，我们还对caspase-3、caspase-9、Akt、pAkt、高迁移率族蛋白B1（HMGB1）、微管相关蛋白1轻链3I（LC3I）、微管相关蛋白1轻链3II（LC3II）、p62、Beclin1以及与凋亡和自噬现象相关的蛋白质进行了表达分析。两名兽医患者使用EP + MMC组合控制角膜鳞状癌切除后的切缘，我们的结果得到了他们的证实。总之，我们确认EP增强MMC治疗效果的原因是抑制了药物诱导的自噬过程，从而有利于细胞凋亡。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/cf6e/8345561/adac1f0a65c9/cancers-13-03867-g007.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/cf6e/8345561/93f57ffec49e/cancers-13-03867-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/cf6e/8345561/88155828f8b4/cancers-13-03867-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/cf6e/8345561/4d8fc692f574/cancers-13-03867-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/cf6e/8345561/31db53688c1b/cancers-13-03867-g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/cf6e/8345561/bf2ab4e7616c/cancers-13-03867-g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/cf6e/8345561/c825fb91d25d/cancers-13-03867-g006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/cf6e/8345561/adac1f0a65c9/cancers-13-03867-g007.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/cf6e/8345561/93f57ffec49e/cancers-13-03867-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/cf6e/8345561/88155828f8b4/cancers-13-03867-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/cf6e/8345561/4d8fc692f574/cancers-13-03867-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/cf6e/8345561/31db53688c1b/cancers-13-03867-g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/cf6e/8345561/bf2ab4e7616c/cancers-13-03867-g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/cf6e/8345561/c825fb91d25d/cancers-13-03867-g006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/cf6e/8345561/adac1f0a65c9/cancers-13-03867-g007.jpg

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本文引用的文献

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Influence of lysosomal sequestration on multidrug resistance in cancer cells.溶酶体隔离对癌细胞多药耐药性的影响。
Cancer Drug Resist. 2019 Mar 19;2(1):31-42. doi: 10.20517/cdr.2018.23. eCollection 2019.
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Drug resistance and combating drug resistance in cancer.癌症中的耐药性与抗耐药性
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Regulation of Autophagy by Glycolysis in Cancer.癌症中糖酵解对自噬的调控
电化学疗法：改善耐药实体瘤治疗的替代策略。
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The application of point source electroporation and chemotherapy for the treatment of glioma: a randomized controlled rat study.点源电穿孔和化疗在治疗脑胶质瘤中的应用：一项随机对照大鼠研究。
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Autophagy and autophagy-related proteins in cancer.癌症中的自噬和自噬相关蛋白。
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Definitive chemoradiotherapy in patients with squamous cell cancers of the head and neck - results from an unselected cohort of the clinical cooperation group "Personalized Radiotherapy in Head and Neck Cancer".根治性放化疗在头颈部鳞癌患者中的应用——来自临床合作组“头颈部癌症个体化放疗”的未经选择队列的结果。
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Quantitative proteomic analysis of enhanced cellular effects of electrochemotherapy with Cisplatin in triple-negative breast cancer cells.电化学治疗联合顺铂增强三阴性乳腺癌细胞疗效的定量蛋白质组学分析。
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