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山羊疱疹病毒 1(CpHV-1)作为溶瘤病毒治疗的潜在候选物。

Caprine herpesvirus 1 (CpHV-1) as a potential candidate for oncolytic virotherapy.

机构信息

a Department of Veterinary Medicine and Animal Productions , University of Naples "Federico II" , Napoli , Italia.

b Department of Pharmacy , University of Salerno , Fisciano (Na) , Italia.

出版信息

Cancer Biol Ther. 2019;20(1):42-51. doi: 10.1080/15384047.2018.1504722. Epub 2018 Nov 8.

Abstract

Caprine Herpesvirus type 1 (CpHV-1) is a species-specific herpes virus able to induce apoptosis in several biological systems. In the present study we aimed to investigate the ability of CpHV-1 to reduce cells viability, to replicate and to cause cell death also in human cancer cell lines. We tested the CpHV-1 effects on HEL-299, Vero, MDA-MB-468, HeLa, U2OS, PC3, A549 and K562 neoplastic cell lines and on MDBK cells. Firstly, we evaluated the effect of CpHV-1 infection on cell viability by MTT assay and our data showed that CpHV-1 can induce a marked cytopathic effect (CPE) in most of cell lines tested, except for HEL-299, Vero and K562 cells. The reduction of cell viability was associated with a significant increase of viral production. We next investigated if CpHV-1 was able to induce cell death and so through western blotting analysis we evaluated cleaved caspase 3, LC3II and p62 protein levels after infection. Caspase 3 activation was detected in MDBK cells and, even if at different times p.i., also in MDA-MB-468, U2OS, and PC3 cell lines, while LC3II increase and concomitant p62 protein reduction were observed only in U2OS, and A549 cells, no significant alteration of these proteins was observed in the other cell lines tested. Finally, to confirm virus ability to trigger apoptosis we performed an Annexin-V apoptosis test after 24 h p.i. Although we need to further explore mechanisms underlying CpHV-1 treatment, this study could serve as the basis for the development of new treatment options aiming to fight several cancer types.

摘要

山羊疱疹病毒 1 型(CpHV-1)是一种具有种属特异性的疱疹病毒,能够在多个生物系统中诱导细胞凋亡。在本研究中,我们旨在研究 CpHV-1 降低细胞活力、复制和引起人类癌细胞系细胞死亡的能力。我们测试了 CpHV-1 对 HEL-299、Vero、MDA-MB-468、HeLa、U2OS、PC3、A549 和 K562 肿瘤细胞系以及 MDBK 细胞的影响。首先,我们通过 MTT 测定法评估了 CpHV-1 感染对细胞活力的影响,我们的数据表明,CpHV-1 可以在大多数测试的细胞系中引起明显的致细胞病变效应(CPE),除了 HEL-299、Vero 和 K562 细胞。细胞活力的降低与病毒产量的显著增加相关。我们接下来研究了 CpHV-1 是否能够诱导细胞死亡,因此通过 Western blot 分析,我们在感染后评估了 cleaved caspase 3、LC3II 和 p62 蛋白水平。在 MDBK 细胞中检测到 caspase 3 的激活,并且即使在不同的感染后时间点,在 MDA-MB-468、U2OS 和 PC3 细胞系中也检测到 caspase 3 的激活,而 LC3II 的增加和同时的 p62 蛋白减少仅在 U2OS 和 A549 细胞中观察到,在其他测试的细胞系中未观察到这些蛋白质的明显变化。最后,为了确认病毒触发细胞凋亡的能力,我们在感染后 24 小时进行了 Annexin-V 凋亡试验。尽管我们需要进一步探索 CpHV-1 治疗的机制,但这项研究可以为开发旨在治疗多种癌症类型的新治疗方法提供基础。

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