胃硬癌细胞通过抑制成纤维细胞自噬诱导其衰老的机制  

Scirrhous Gastric Cancer Cell Induces Senescence by Inhibiting Autophagy of Fibroblasts

作　　者：韩方 姜飞 张宇华 张成武 HAN Fang;JIANG Fei;ZHANG Yuhua;ZHANG Chengwu(Department of Hepatobiliary Pancreatic Surgery,Minimal Invasive Surgery,Zhejiang Provincial People’s Hospital,Hangzhou Medical College,Hangzhou 310014,China;Clinical Laboratory Center,Zhejiang Provincial People’s Hospital,Hangzhou Medical College,Hangzhou 310014,China)

机构地区：[1]杭州医学院附属浙江省人民医院肝胆胰微创外科,杭州310014 [2]杭州医学院附属浙江省人民医院检验中心,杭州310014

出　　处：《中国医科大学学报》2018年第11期985-988,998,共5页Journal of China Medical University

基　　金：浙江省自然科学基金(LY15H160054)

摘　　要：目的检测胃硬癌的条件培养基(CM)对成纤维细胞自噬和衰老的影响,探讨胃硬癌细胞对成纤维细胞的影响及机制。方法利用胃硬癌细胞株KATOⅢ和NUGC4的CM作用于人正常成纤维细胞系FEF3和WI38。共培养并观察其形态学改变,检测增殖情况。Western blotting检测凋亡、自噬、衰老相关蛋白的变化。SA-β-Gal染色研究成纤维细胞衰老情况。检测雷帕霉素和CM作用下对成纤维细胞衰老和自噬的影响。结果 CM作用下成纤维细胞出现了细胞质空泡化、核固缩等特征性改变,同时自噬被抑制。随着作用时间增加,成纤维细胞数量减少,衰老相关蛋白p21表达上调。SA-β-Gal染色后残存的细胞表现衰老的特征。采用雷帕霉素处理成纤维细胞诱导自噬后,自噬被加强,而衰老被抑制。结论胃硬癌细胞通过抑制成纤维细胞的自噬,诱导成纤维细胞提前衰老。Objective To investigate the effects of conditioned medium(CM)of scirrhous gastric cancer cell on autophagy and senescence of fibroblasts.Methods The normal human fibroblast cell lines FEF3 and WI38 were treated with CM of scirrhous gastric cancer cell lines KATOⅢand NUGC4.Morphological changes were observed after 48 h,and the changes in the levels of proteins related to apoptosis,autophagy,and senescence were detected using Western blotting.SA-β-Gal staining was used to detect the senescence of fibroblasts.Rapamycin was used to induce autophagy.Results The fibroblasts cultured with CM showed morphological changes such as cytoplasmic vacuolation and nuclear pyknosis,and autophagy was inhibited.The proliferation assay demonstrated a decreased number of the fibroblasts treated with CM.Meanwhile,autophagy was inhibited,and senescence was upregulated.After SA-β-Gal staining,the remaining cells were characterized by premature senescence.However,autophagy was restored,and senescence status was blocked after treatment of the fibroblasts with rapamycin.Conclusion Scirrhous gastric cancer cells could induce premature senescence by inhibiting autophagy of fibroblasts.

关 键 词：胃硬癌 成纤维细胞 自噬 衰老 条件培养基 

分 类 号：R73-37[医药卫生—肿瘤]