摘要
目的研究新城疫病毒(newcastle disease virus,NDV)毒株FMW(NDV/FMW)诱导结肠癌Caco2细胞发生凋亡及其机制。方法通过病毒滴度法测定病毒在细胞内复制情况;CCK8法检测细胞存活率;显微镜下观察细胞形态学的变化;Western印迹法检测细胞凋亡相关蛋白Caspase3、PARP及MAPK下游信号通路相关p38、JNK、Erk1/2蛋白表达水平。结果NDV/FMW在Caco2细胞内复制,降低2D及3D培养Caco2细胞存活率(P<0.001);NDV/FMW诱导Caco2细胞中裂解的Caspase3(cleaved-Caspase3)及PARP裂解片段(cleaved-PARP)增加,泛Caspase抑制剂预处理降低Caspase3及PARP裂解片段表达水平;NDV/FMW感染组Caco2细胞磷酸化p38和磷酸化JNK较对照组增加,磷酸化ERK1/2及总p38、JNK、ERK1/2蛋白水平无显著增加;与NDV/FMW感染组相比较,p38抑制剂预处理组凋亡相关蛋白表达水平显著减低,但JNK抑制剂预处理组凋亡相关蛋白表达水平没有显著变化。结论NDV/FMW诱导结肠癌Caco2细胞发生Caspase和p38MAPK依赖性细胞凋亡。
Objective To investigate the effect of Newcastle disease virus(NDV)in induction of apoptosis in colon cancer cell line Caco2 and its mechanisms.Methods Human colon cancer Caco2 cells were infected with Newcastle disease virus FMW strain(NDV/FMW),the virus yield was detected with multi-step viral growth curve.Cell viability was detected by CCK8;the cell morphological change was observed by microscope;the changes of apoptosis-related proteins expression including Caspase3,PARP(poly ADP-ribose polymerase)and p38,JNK,Erk1/2,downstream of MAPK signaling pathway were detected by Western blotting.Results NDV/FMW replicated in Caco2 cells,inhibited cell viability in two-dimensional(2D)and three-dimensional(3D)cell cultures(P<0.001);NDV/FMW increased Caspase3(cleaved-Caspase3)and PARP cleavage(cleaved-PARP)in Caco2 cells.Caspase inhibition reduced the expression of Caspase3 and PARP fragment;NDV/FMW infection resulted the increased expression of phosphorylated p38 and JNK in Caco2 cells,while the phosphorylated ERK1/2 and total p38,JNK,ERK1/2 protein levels did not increase significantly;compared with NDV/FMW infected group,the expression levels of apoptosis related protein decreased significantly in p38 inhibitor group,but the apoptosis related protein in JNK inhibitor did not change significantly.Conclusion NDV/FMW can induce Caspase and p38MAPK-dependent apoptosis in Caco2 colon cancer cells.
作者
古丽娜儿
王思橦
黄莉莉
刘竹青
朱忠政
许青
Gulinaer;WANG Si-tong;HUANG Li-li;LIU Zhu-qing;ZHU Zhong-zheng;XU Qing(Dept.of Oncology,Tenth People's Hospital,Tongji University School of Medicine,Shanghai 200072,China;Dept.of Oncology,Shanghai Dermatology Hospital,Shanghai 200433,China;Cancer Center of Tongji University,Shanghai 200072,China)
出处
《同济大学学报（医学版）》
CAS
2020年第5期546-551,共6页
Journal of Tongji University(Medical Science)
基金
上海市科学技术委员会项目(18DZ1910102)
上海市卫生和计划生育委员会项目(2016LP029)
上海市卫生计生委重点项目(201640020)
上海市申康临床三年行动计划项目(16CR3073B)
上海市科委西医引导项目(17411967300)。
