核心蛋白聚糖在胰腺癌beta细胞功能中的作用及对线粒体分离融合的影响  

The role of decorin in the function of pancreatic cancer beta cells and its influence on the separation and fusion of mitochondria

作　　者：张荣 吕海龙 杨一邨[1] 王浩斌 黄江涛[1] 张抒[1] ZHANG Rong;LVU Hailong;YANG Yicun;WANG Haobin;HUANG Jiangtao;ZHANG Shu(Department of General Surgery,the Third People′s Hospital of Chengdu,Chengdu,Sichuan 610031,China)

机构地区：[1]四川省成都市第三人民医院普外科,610031

出　　处：《重庆医学》2022年第8期1272-1278,1284,共8页Chongqing medicine

基　　金：国家自然科学基金项目(81702963)。

摘　　要：目的探究核心蛋白聚糖(DCN)在胰腺癌组织中的表达及对胰岛beta细胞线粒体分离融合的影响。方法征集21例胰腺导管腺癌(PDAC组)、25例胰腺神经内分泌肿瘤(PNET组)和4例健康者(健康组)胰腺组织;免疫组织化学检测胰腺组织中DCN表达;免疫荧光法检测胰腺组织中线粒体融合蛋白2(MFN2)和动力相关蛋白1(DRP1)表达情况;蛋白免疫印迹(Western blot)法检测胰腺组织中DCN、线粒体分裂因子(MFF)、MFN1、MFN2、视神经萎缩症蛋白1(OPA1)、DRP1和线粒体分裂蛋白1(FIS1)表达;CCK-8法检测正常糖培养条件下DCN(0、1、10、100和200 nmol/L)处理对人胰岛β细胞(0、12、24和48 h)增殖的影响;胰岛β细胞分为2组:正常糖和高糖(HG)培养,均分别采用0 nmol/L和100 nmol/L DCN处理细胞24 h,CCK-8法再次检测各组细胞增殖,Annexin V/PI染色检测各组细胞凋亡,酶联免疫吸附实验检测各组细胞培养上清液中胰岛素分泌量,Western blot法检测MFF、MFN1、MFN2、OPA1、DRP1、FIS1、烟酰胺腺嘌呤二核苷酸磷酸氧化酶(P22^(phox))、磷酸化丝裂原活化蛋白激酶p38亚基(p-p38)、p38、胰岛素样生长因子Ⅰ受体蛋白(IGFIR)、磷酸化蛋白激酶B(p-AKT)和AKT的表达。结果与健康组比较,PDAC和PNET组癌组织中DCN、DRP1、FIS1和MFF表达水平明显降低,MFN1、MFN2和OPA表达水平明显升高(P<0.05);在正常糖条件下,100 nmol/L和200 nmol/L DCN处理24 h和48 h的细胞增殖倍数均明显高于对应时间点的0 nmol/L DCN组,且具有时间和剂量依赖性(P<0.05);在正常糖和HG条件下,100 nmol/L DCN组的细胞增殖倍数均明显高于0 nmol/L DCN组(P<0.05);在正常糖和HG条件下,100 nmol/L DCN组的AnnexinV^(+)PI^(-)细胞比例较0 nmol/L DCN组无明显变化(P>0.05);无论正常糖或是HG条件下,0 nmol/L DCN组IGFIR表达水平与100 nmol/L DCN组比较,差异均无统计学意义(P>0.05);在正常糖条件下,100 nmol/L DCN组的培养上清液中胰岛素分泌量,细胞中DRP1、FIS1、Objective To explore the expression of decorin(DCN)in pancreatic cancer tissues and its effect on the mitochondrial fission and fusion of pancreatic beta cells.Methods Pancreatic tissues were collected from 21 case of pancreatic ductal adenocarcinoma(the PDAC group),25 case of pancreatic neuroendocrine tumors(the PNET group)and 4 case of healthy persons(the healthy group).The expression of DCN in pancreatic tissue was detected by immunohistochemistry and Western blot.The protein expression levels of mitochondrialfusion protein 2(MFN2)and dynamo-related protein 1(DRP1)in pancreatic tissue were detected by immunofluorescence.The protein expression levels of DCN,mitochondrial fission factor(MFF),MFN1,MFN2,optic atrophy protein 1(OPA1),DRP1 and mitochondrial fission protein 1(FIS1)in pancreatic tissue were detected by Western blot.CCK-8 method was used to detect the effects of DCN(0,1,10,100 and 200 nmol/L)on the proliferation of human pancreaticβcells(0,12,24 and 48 h)under normal glucose culture conditions.Pancreaticβcells were divided into two groups:the normal glucose group and the high glucose group,treated with 0 nmol/L and 100 nmol/L DCN for 24 h,respectively.The proliferation of human pancreaticβcells was detected again by CCK-8 method.The apoptosis was detected by Annexin V/PI staining.Insulin secretion was detected by enzyme-linked immunosorbent assay.The protein expression levels of MFF,MFN1,MFN2,OPA1,DRP1,FIS1,nicotinamide adenine dinucleotide phosphate oxidase(P22^(phox)),phosphorylated mitogen-activated protein kinase P38 subunit(p-p38),p38,insulin-like growth factorⅠreceptor protein(IGFIR),phosphate expression of protein kinase B(p-AKT)and AKT were detected by Western blot.Results Compared with the healthy group,the expression levels of DCN,DRP1,FIS1 and MFF in the cancer tissues of the PDAC and the PNET groups were significantly decreased,and the expression levels of MFN1,MFN2 and OPA were significantly increased(P<0.05).Under normal glucose conditions,the proliferation of pancreaticβcells treat

关 键 词：胰腺癌 核心蛋白聚糖 胰岛β细胞 线粒体分裂 线粒体融合 P22^(phox)/MAPK通路 

分 类 号：R576[医药卫生—消化系统]