Gao Hui, Li Ling Yu, Zhang Man, Zhang Quan
College of Clinical Medicine of Yangzhou University, Yangzhou 225001, Jiangsu, China; Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, College of Veterinary Medicine, Yangzhou University, Yangzhou 225009, Jiangsu, China.
Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, College of Veterinary Medicine, Yangzhou University, Yangzhou 225009, Jiangsu, China.
Biomed Environ Sci. 2016 Dec;29(12):877-884. doi: 10.3967/bes2016.117.
This paper aims to investigate the apoptotic effect of inactivated Sendai virus (hemagglutinating virus of Japan-enveloped, HVJ-E) on murine melanoma cells (B16F10) and the possible mechanisms involved in the putative apoptotic reactions.
B16F10 cells were treated with HVJ-E at various multiplicities of infection (MOI), and the reactive oxygen species (ROS), cell viability, and apoptosis were measured. Next, the roles of ROS in the regulation of Bcl-2/Bax and the activation of mitogen-activated protein kinase (MAPK) pathways in HVJ-E-treated B16F10 cells were analyzed. To further evaluate the cytotoxic effect of HVJ-E-generated ROS on B16F10 cells, HVJ-E was intratumorally injected, both with and without N-acetyl-L-cysteine (NAC), into melanoma tumors on BALB/c mice. Tumor volume was then monitored for 3 weeks, and the tumor proteins were separated for immunoblot assay.
Treatment of B16F10 cells with HVJ-E resulted in a dose-dependent inhibition of cell-viability and an induction of apoptosis. The latter effect was associated with the generation of ROS. Inhibition of ROS generation by NAC resulted in a significant reduction of HVJ-E-induced Erk1/2, JNK, and p38 MAPK activation. Additionally, ROS inhibition caused a decrease in the Bcl-2/Bax ratio as well as promoting activation of apoptosis both in vitro and in vivo.
These results suggest that HVJ-E possesses potential anticancer activity in B16F10 cells through ROS-mediated mitochondrial dysfunction involving the MAPK pathway.
本文旨在研究灭活仙台病毒（日本血凝病毒包膜型，HVJ-E）对小鼠黑色素瘤细胞（B16F10）的凋亡作用以及推测的凋亡反应中涉及的可能机制。
用不同感染复数（MOI）的HVJ-E处理B16F10细胞，检测活性氧（ROS）、细胞活力和凋亡情况。接下来，分析ROS在HVJ-E处理的B16F10细胞中对Bcl-2/Bax调节以及丝裂原活化蛋白激酶（MAPK）途径激活中的作用。为进一步评估HVJ-E产生的ROS对B16F10细胞的细胞毒性作用，将HVJ-E瘤内注射到BALB/c小鼠的黑色素瘤肿瘤中，注射时分别添加和不添加N-乙酰-L-半胱氨酸（NAC）。然后监测肿瘤体积3周，并分离肿瘤蛋白进行免疫印迹分析。
用HVJ-E处理B16F10细胞导致细胞活力呈剂量依赖性抑制并诱导凋亡。后一种作用与ROS的产生有关。NAC抑制ROS产生导致HVJ-E诱导的Erk1/2、JNK和p38 MAPK激活显著降低。此外，ROS抑制导致Bcl-2/Bax比值降低，并在体外和体内均促进凋亡激活。
这些结果表明，HVJ-E通过ROS介导的涉及MAPK途径的线粒体功能障碍在B16F10细胞中具有潜在的抗癌活性。
