Department of Microbiology, Faculty of Biotechnology and Biomolecular Sciences, Universiti Putra Malaysia, UPM, 43400, Serdang, Selangor Darul Ehsan, Malaysia.
Malaysia Genome Institute, Ministry of Science, Technology and Innovation, Jalan Bangi, 43000, Kajang, Selangor Darul Ehsan, Malaysia.
BMC Cancer. 2021 May 27;21(1):625. doi: 10.1186/s12885-021-08345-y.
Newcastle disease virus (NDV) is an oncolytic virus with excellent selectivity against cancer cells, both in vitro and in vivo. Unfortunately, prolonged in vitro NDV infection results in the development of persistent infection in the cancer cells which are then able to resist NDV-mediated oncolysis. However, the mechanism of persistency of infection remains poorly understood.
In this study, we established persistently NDV-infected EJ28 bladder cancer cells, designated as EJ28P. Global transcriptomic analysis was subsequently carried out by microarray analysis. Differentially expressed genes (DEGs) between EJ28 and EJ28P cells identified by the edgeR program were further analysed by Gene Set Enrichment Analysis (GSEA) and Ingenuity Pathway Analysis (IPA) analyses. In addition, the microarray data were validated by RT-qPCR.
Persistently NDV-infected EJ28 bladder cancer cells were successfully established and confirmed by flow cytometry. Microarray analysis identified a total of 368 genes as differentially expressed in EJ28P cells when compared to the non-infected EJ28 cells. GSEA revealed that the Wnt/β-catenin and KRAS signalling pathways were upregulated while the TGF-β signalling pathway was downregulated. Findings from this study suggest that the upregulation of genes that are associated with cell growth, pro-survival, and anti-apoptosis may explain the survivability of EJ28P cells and the development of persistent infection of NDV.
This study provides insights into the transcriptomic changes that occur and the specific signalling pathways that are potentially involved in the development and maintenance of NDV persistency of infection in bladder cancer cells. These findings warrant further investigation and is crucial towards the development of effective NDV oncolytic therapy against cancer.
新城疫病毒（NDV）是一种溶瘤病毒，对体外和体内的癌细胞具有极好的选择性。不幸的是，NDV 的体外长期感染会导致癌细胞中持续性感染的发展，从而使癌细胞能够抵抗 NDV 介导的溶瘤作用。然而，持续性感染的机制仍知之甚少。
在本研究中，我们建立了持续性 NDV 感染的 EJ28 膀胱癌细胞，命名为 EJ28P。随后通过微阵列分析进行了全局转录组分析。通过 edgeR 程序鉴定的 EJ28 和 EJ28P 细胞之间的差异表达基因（DEGs）进一步通过基因集富集分析（GSEA）和 Ingenuity 通路分析（IPA）进行分析。此外，通过 RT-qPCR 验证了微阵列数据。
成功建立了持续性 NDV 感染的 EJ28 膀胱癌细胞，并通过流式细胞术进行了验证。微阵列分析鉴定出 EJ28P 细胞与未感染的 EJ28 细胞相比共有 368 个基因差异表达。GSEA 结果显示，Wnt/β-catenin 和 KRAS 信号通路上调，而 TGF-β 信号通路下调。本研究结果表明，与细胞生长、生存和抗凋亡相关的基因上调可能解释了 EJ28P 细胞的生存能力和 NDV 持续性感染的发展。
本研究提供了对发生的转录组变化的深入了解，以及可能参与膀胱癌细胞中 NDV 持续性感染的发展和维持的特定信号通路。这些发现值得进一步研究，对于开发有效的 NDV 溶瘤疗法治疗癌症至关重要。
