Abstract

A test of the sensitivity of seven colon cancer cell lines to a panel of 12 nonpathogenic human enteroviruses revealed significant differences in the ability of tumor cells to become infected and replicate different viral strains. Among the factors that can affect the sensitivity of cells to viruses are differences in the state of the mechanisms of antiviral protection, associated with a reaction to type I interferons. Using the two colon cancer cell lines CaCo2 and LIM1215 as a model, significant differences were revealed in the ability of cells to defend themselves against virus infection after 16 hours of treatment with 1000 units/mL of interferon-alpha. To study the effect of the state of the interferon response system, represented by the Jak/STAT signaling pathway, on the sensitivity of cells to different strains of enteroviruses, HEK293T cell lines were used. These are capable of supporting replication of each of the tested enteroviruses, as well as maintaining the ability to protect against viral infection after the treatment with interferon. Using the CRISPR/Cas9 system, HEK293T sublines with knockouts of the IFNAR1 and STAT2 genes were obtained. The sensitivity of control and knockout cells to infection with five strains of enteroviruses and the vesicular stomatitis virus was analyzed. It was noted that knockout of the IFNAR1 and STAT2 genes resulted in an increased sensitivity to all tested viruses. In knockout cells, the levels of reproduction of the vaccine dervied of poliovirus type 1, Echoviruses 7 and 30, and Coxsackie viruses B5 and A7 were also significantly increased in comparison with the control HEK293T cells. Thus, deficiencies in the Jak/STAT signaling pathway in tumor cells lead to an overall increase in the sensitivity to oncolytic viruses.

中文翻译：

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Jak / Stat通路的状态影响肿瘤细胞对溶瘤性肠病毒的敏感性

摘要

对七个结肠癌细胞系对一组12种非致病性人肠病毒的敏感性的测试显示，肿瘤细胞被感染和复制不同病毒株的能力显着不同。在可能影响细胞对病毒敏感性的因素中，与对I型干扰素的反应有关的抗病毒保护机制的状态存在差异。使用两种结肠癌细胞系CaCo2和LIM1215作为模型，在用1000单位/ mL的干扰素-α治疗16小时后，细胞防御病毒感染的能力显着不同。为了研究以Jak / STAT信号通路为代表的干扰素应答系统状态对细胞对不同株肠病毒的敏感性的影响，使用了HEK293T细胞系。这些能够支持每种被测试的肠病毒的复制，以及在用干扰素治疗后保持防止病毒感染的能力。使用CRISPR / Cas9系统，HEK293T与获得了IFNAR1和STAT2基因。分析了对照细胞和敲除细胞对五株肠病毒和水泡性口炎病毒感染的敏感性。注意到敲除IFNAR1和STAT2基因导致对所有测试病毒的敏感性增加。在敲除细胞中，与脊髓灰质炎病毒1型，脊髓灰质炎7型和30型脊髓灰质炎病毒以及柯萨奇B5和A7型脊髓灰质炎病毒相比，衍生的疫苗繁殖水平与对照HEK293T细胞相比也显着提高。因此，肿瘤细胞中Jak / STAT信号传导途径的缺陷导致对溶瘤病毒的敏感性的总体提高。