DNER通过抑制线粒体自噬促进胃癌细胞恶性进展的机制研究  

Study on the Mechanism of DNER Promoting Malignant Progression of Gastric Cancer Cells by Inhibiting Mitochondrial Autophagy

作　　者：付永生 卢静芬 赵昕 王卫 朱其聪 FU Yongsheng;LU Jingfen;ZHAO Xin;WANG Wei;ZHU Qicong(Department of Hematology and Oncology,928 Medical Hospital of PLA Joint Logistic Support Force,Haikou 570100,China;Department of General Surgery,928 Medical Hospital of PLA Joint Logistic Support Force,Haikou 570100,China;Department of General Surgery,the Third People’s Hospital of Haikou,Haikou 570100,China)

机构地区：[1]解放军联勤保障部队第九二八医院血液肿瘤科,海口570100 [2]解放军联勤保障部队第九二八医院普通外科,海口570100 [3]海口市第三人民医院普通外科,海口570100

出　　处：《现代检验医学杂志》2024年第4期50-55,共6页Journal of Modern Laboratory Medicine

基　　金：海南省卫生厅科学研究项目(编号:22A200057)。

摘　　要：目的研究Delta/Notch样表皮生长因子相关受体(delta/notch-like epidermal growth factor-related receptor,DNER)在胃癌中的作用及其调节机制。方法通过实时定量聚合酶链反应(qRT-PCR)和蛋白免疫印迹(Western blot)检测胃癌组织和细胞中DNER mRNA和蛋白表达水平。构建沉默DNER表达的胃癌细胞系SGC7901,用线粒体动力相关蛋白(dynamin-related protein 1,DRP1)抑制剂Mdivi-1处理细胞。CCK-8法、Transwell实验和流式细胞术分别检测细胞活力、侵袭能力和细胞凋亡。Western blot检测DNER蛋白、凋亡相关蛋白[半胱天冬氨酸蛋白酶3(cysteinyl aspartate-specific proteinase-3,Caspase-3)、Bcl-2相关X蛋白(Bcl-2 associated X,Bax)]、自噬相关蛋白[微管相关蛋白1轻链3-Ⅱ/Ⅰ(microtubule-associated protein 1 light chain 3-II/Ⅰ,LC3Ⅱ/Ⅰ)、p62,PTEN诱导激酶1(PTEN induced putative kinase 1,PINK1)和Parkin],以及线粒体裂变和融合蛋白[DRP1,线粒体分裂因子(mitochondrial fission factor,MFF)、线粒体分裂蛋白1(fission mitochondrial 1,FIS1)、视神经萎缩蛋白1(optic atrophy 1,OPA1)、线粒体融合蛋白1(mitofusin 1,MFN1)和MFN2]水平。结果胃癌肿瘤组织和细胞中DNER mRNA,蛋白表达水平分别显著高于相邻正常组织(t=-52.485,-46.955)和人正常胃上皮细胞(F=60.551,60.652),差异具有统计学意义(均P<0.001)。沉默DNER显著抑制SGC7901细胞的增殖和侵袭、诱导细胞凋亡、增加细胞凋亡相关蛋白表达,差异具有统计学意义(t=8.026~25.903,均P<0.05)。沉默DNER显著升高LC3Ⅱ/Ⅰ比率(t=18.086),降低p62蛋白水平(t=6.747),促进PINK1和Parkin蛋白在线粒体的聚集(t=15.630,18.171),抑制线粒体融合蛋白OPA1,MFN1和MFN2表达(t=12.835,8.963,9.732),促进线粒体裂变蛋白DRP1,MFF和FIS1表达(t=16.034,16.939,15.971),差异具有统计学意义(均P<0.05)。Mdivi-1处理可抵消沉默DNER对胃癌细胞线粒体自噬及细胞增殖、侵袭和凋亡的影响。结论DNER通过抑制线粒体动力学失衡减�Objective To investigate the role of delta/notch-like epidermal growth factor-related receptor(DNER)in gastric cancer and its regulatory mechanism.Methods The mRNA and protein levels of DNER in gastric cancer tissues and cells were detected with quantitative real time polymerase chain reaction(qRT-PCR)and Western blot.Gastric cancer cell line SGC7901 with silenced DNER expression was constructed,and cells were treated with mitochondrial dynamin-related protein 1(DRP1)inhibitor Mdivi-1.CCK-8 assay,Transwell assay,and flow cytometry were used to detect cell viability,invasion ability and apoptosis,respectively.Western blot was used to detect DNER protein levels,apoptosis-associated proteins[Cysteinyl aspartatespecific proteinase-3(Caspase-3),Bcl-2 Associated X(Bax)],autophagy associated proteins[microtubule-associated protein 1 light chain 3-II/Ⅰ,LC3Ⅱ/Ⅰ),p62,PTEN induced putative kinase 1(PINK1)and Parkin],and mitochondrial fission and fusion protein[DRP1,mitochondrial fission factor(MFF),mitochondrial fission protein 1(FIS1),Optic Atrophy 1(OPA1),mitofusin 1(MFN1)and MFN2]levels.Results The expression levels of DNER mRNA and protein in gastric cancer tissues were higher than those in adjacent normal tissues(t=-52.485,-46.955),while expression levels of DNER mRNA and protein in gastric cancer cells were higher than those in normal gastric epithelial cells(F=60.551,60.652),and the differences were significant(P<0.001).Silencing DNER inhibited the proliferation and invasion of SGC7901 cells,induced apoptosis,and increased the expression of apoptosis-related proteins,with significant differences(t=8.026~25.903,all P<0.05).Silenced DNER increased LC3Ⅱ/Ⅰratio(t=18.086),decreased p62 protein level(t=6.747),promoted the aggregation of PINK1 and Parkin proteins in mitochondria(t=15.630,18.171),inhibited the expression of mitochondrial fusion proteins OPA1,MFN1 and MFN2(t=12.835,8.963,9.732),and promoted the expression of mitochondrial fission proteins DRP1,MFF and FIS1(t=16.034,16.939,15.971),with significant dif

关 键 词：胃癌 自噬 Delta/Notch样表皮生长因子相关受体 线粒体融合 线粒体裂变 

分 类 号：R735.2[医药卫生—肿瘤] R730.43[医药卫生—临床医学]