Xia Mao, Meng Gang, Dong Jie
Department of Laboratory Medicine, The Affiliated Drum Tower Hospital, Medical School of Nanjing University, Nanjing 210007, China.
Zhongguo Fei Ai Za Zhi. 2021 Apr 20;24(4):245-253. doi: 10.3779/j.issn.1009-3419.2021.102.14. Epub 2021 Mar 29.
Lung cancer is one of the leading causes of cancer-related morbidity and mortality. Oncolytic virotherapy is an emerging therapeutic modality that utilizes replication-competent viruses to destroy cancers. As a powerful tool to kill tumor cells with excellent safety profile, attenuated measles virus of the Edmonston strain (MV-Edm) has been widely applied in the development of tumor therapy and preclinical trials. The aim of this study was to investigate the synergistic effect of nuclear factor kappa B (NF-κB) signaling pathway inhibitor and oncolytic measles virus vaccine against lung cancer and the involved mechanisms.
Using Western blot to detect MV-Edm infection of A549 and H1299 were infected by MV-Edm alone or used the NF-κB pathway inhibitor PS1145/cell autophagy related siRNA, expression level of p-IκBα, IκBα, PARP and BAX were determined by western blot. Using flow cytometry to analysis the rate of apoptosis, and using MTT [3-(4,5)-dimethylthiahiazo(-z-y1)-3,5-di-phenytetrazoliumromide] method to detect the cell survival rate.
Inhibition of cell autophagy could obviously inhibit the MV-Edm infection induced the NF-κB pathway activation in A549 and H1299. In MV-Edm infected A549 and H1299, p-IκBα level increased and IκBα level decreased over infection time, compared with control group. Inhibition of the NF-κB pathway by PS1145 could promote the apoptosis of MV-Edm infected A549 and H1299 and amplify the tumor killing effect.
The combination of NF-κB signaling pathway inhibitor pS1145 and oncolytic measles virus vaccine strains can promote the apoptosis of human lung cancer cells A549 and H1299 and enhance their oncolytic effect.
肺癌是癌症相关发病和死亡的主要原因之一。溶瘤病毒疗法是一种新兴的治疗方式，它利用具有复制能力的病毒来摧毁癌症。作为一种具有出色安全性的杀死肿瘤细胞的强大工具，埃德蒙斯顿株减毒麻疹病毒（MV-Edm）已广泛应用于肿瘤治疗的开发和临床前试验。本研究的目的是探讨核因子κB（NF-κB）信号通路抑制剂与溶瘤麻疹病毒疫苗联合抗肺癌的协同作用及其相关机制。
采用蛋白质免疫印迹法检测单独用MV-Edm或使用NF-κB通路抑制剂PS1145/细胞自噬相关小干扰RNA（siRNA）感染A549和H1299细胞后，通过蛋白质免疫印迹法测定磷酸化IκBα（p-IκBα）、IκBα、聚（ADP-核糖）聚合酶（PARP）和凋亡相关蛋白BAX的表达水平。采用流式细胞术分析细胞凋亡率，并用噻唑蓝（MTT）法检测细胞存活率。
抑制细胞自噬可明显抑制MV-Edm感染诱导的A549和H1299细胞中NF-κB通路的激活。在MV-Edm感染的A549和H1299细胞中，与对照组相比，随着感染时间的延长，p-IκBα水平升高，IκBα水平降低。PS1145抑制NF-κB通路可促进MV-Edm感染的A549和H1299细胞凋亡，并增强肿瘤杀伤作用。
NF-κB信号通路抑制剂pS1145与溶瘤麻疹病毒疫苗株联合应用可促进人肺癌细胞A549和H1299凋亡并增强其溶瘤效果。
