中国肿瘤生物治疗杂志2026，Vol.33Issue(2)：132-139,8.DOI:10.3872/j.issn.1007-385x.2026.02.003
干扰素刺激基因家族成员黏病毒抵抗蛋白2限制肿瘤细胞对呼肠孤病毒溶瘤敏感性的实验研究
Experimental study on interferon-stimulated gene myxovirus resistance protein 2-mediated restriction of tumor cell sensitivity to reovirus oncolysis
摘要
Abstract
Objective:To investigate the role and underlying mechanism of myxovirus resistance protein 2(MX2),a member of the interferon-stimulated gene family,in modulating tumor cell sensitivity to reovirus(Reo)-mediated oncolysis.Methods:Four human tumor cell lines with distinct drug-resistance characteristics were selected,and their sensitivity to Reo-mediated oncolysis was evaluated using the CCK-8 assay.Transcriptome sequencing was performed to identify differentially expressed genes,and MX2 was screened as a candidate gene.The differential expression of MX2 was validated by qPCR and WB assay.In the Reo-low-sensitive COC1/DDP cells,MX2 was knocked down using siRNA,followed by Reo infection,and cell viability was assessed using CCK-8 assay.Viral replication was evaluated by measuring S1 gene expression using qPCR,intracellular viral protein accumulation by immunofluorescence staining,and viral titer by the TCID50 assay.Intracellular Reo dsRNA levels,reactive oxygen species(ROS)production,and apoptosis rates were measured using flow cytometry.Ultrastructural changes in the endoplasmic reticulum(ER)were examined using transmission electron microscopy(TEM).Additionally,WB was used to assess the expression of ER stress-related proteins,including JNK,p-JNK,eIF2α,p-eIF2α,CHOP,and PERK.Results:Among the four tested tumor cell lines,SKOV3 cells exhibited high sensitivity to Reo-mediated oncolysis,whereas COC1/DDP,HuH-7SRB,and SNU-398 cells showed low sensitivity.Transcriptome sequencing revealed that MX2 expression was significantly higher in Reo-low-sensitive cell lines compared with Reo-high-sensitive cells(P＜0.01).In COC1/DDP cells,MX2 knockdown markedly enhanced Reo replication,increased apoptosis,and elevated intracellular ROS levels(all P＜0.001).TEM revealed typical ultrastructural features of ER stress in MX2-knockdown COC1/DDP cells following Reo infection,including ER swelling,dilation,and fragmentation.WB analysis showed significant upregulation of key ER stress markers(p-eIF2α/eIF2α,PERK,and CHOP)and apoptosis-related regulatory proteins JNK/p-JNK(P＜0.05 or P＜0.01).Conclusion:Tumor cell sensitivity to Reo-mediated oncolysis is closely associated with intracellular MX2 expression levels.Knockdown of MX2 significantly enhanced intracellular Reo replication,leading to ROS accumulation,activation of ER stress,and induction of apoptosis.The synergistic effects of enhanced viral replication and apoptosis ultimately potentiate the oncolytic efficacy of Reo.关键词
呼肠孤病毒/黏病毒抵抗蛋白2/病毒复制/活性氧/内质网应激/凋亡/溶瘤效应Key words
reovirus(Reo)/myxovirus resistance protein 2(MX2)/viral replication/reactive oxygen species(ROS)/endoplasmic reticulum stress/apoptosis/oncolytic effect分类
医药卫生引用本文复制引用
梁丹,杨再玲,余佳霓,李欣兰,沈涛,孙永顺,韦永竹,赵星..干扰素刺激基因家族成员黏病毒抵抗蛋白2限制肿瘤细胞对呼肠孤病毒溶瘤敏感性的实验研究[J].中国肿瘤生物治疗杂志,2026,33(2):132-139,8.基金项目
国家自然科学基金(82460604) （82460604）
贵州省科技计划支撑项目资助[黔科合支撑(2025)一般126号] （2025）
贵州医科大学肿瘤免疫治疗工程研究中心项目(校工程中心,第2024[001]号) （校工程中心,第2024[001]号）
