高糖状态通过升高缺氧诱导因子-1α促进胰腺癌的侵袭能力  

High glucose state promotes the invasion of pancreatic cancer by upregulating hypoxia inducible factor-1α

作　　者：秦涛[1] 肖莹 刘晗[2] 黎韡[1] 仵正[1] 王铮[1] 马清涌[1] 马吉光 Qin Tao;Xiao Ying;Liu Han;Li Wei;Wu Zheng;Wang Zheng;Ma Qingyong;Ma Jiguang(Department of Hepatobiliary Surgery,First Affiliated Hospital of Xi′an Jiaotong University,Xi′an 710061,China;Department of Hepatobiliary Surgery,Qilu Hospital of Shandong University,Jinan 250012,China;Department of Anesthesiology,First Affiliated Hospital of Xi'an Jiaotong University,Xi′an 710061,China)

机构地区：[1]西安交通大学第一附属医院肝胆外科,西安710061 [2]山东大学齐鲁医院肝胆外科,济南250012 [3]西安交通大学第一附属医院麻醉科,西安710061

出　　处：《中华胰腺病杂志》2020年第6期418-423,共6页Chinese Journal of Pancreatology

基　　金：国家自然科学基金(81702916);陕西省自然科学基金(2019JQ-959)。

摘　　要：目的探讨缺氧诱导因子-1α(HIF-1α)介导的高血糖微环境对胰腺癌细胞侵袭能力的影响及其机制。方法收集2012年1月至2014年12月间西安交通大学第一附属医院46例行胰腺癌手术切除患者的临床资料,按血糖水平的高低分为血糖正常组(≥3.9 mmol/L~<6.1 mmol/L)和高血糖组(≥6.1 mmol/L),分析两组患者瘤体大小及肿瘤侵袭、转移情况,免疫组织化学法检测肿瘤组织缺氧程度和HIF-1α的表达。构建糖尿病裸鼠胰腺原位移植瘤模型,以连续两次血糖测量结果≥20 mmol/L且尿糖阳性为高血糖裸鼠建模成功,将裸鼠分为血糖正常组和高血糖组,观察胰腺原位移植瘤的大小、缺氧状态及肿瘤侵犯、转移情况。体外培养胰腺癌BxPC3细胞,分为高糖组(用25 mmol/L葡萄糖培养液培养)、缺氧组(应用150μmol/L CoCl2干预)、高糖+缺氧组和对照组(用常规培养液培养);同时采用脂质体法将靶向HIF-1α的siRNA(siRNA-HIF-1α)转染BxPC3细胞,用常规培养液或高糖培养液(含25 mmol/L葡萄糖)培养细胞,分别设为siRNA-HIF-1α组、高糖+siRNA-HIF-1α组,并设阴性对照siRNA转染组(siRNA-NC组)和高糖+siRNA-NC组。采用蛋白质免疫印迹法检测各组细胞HIF-1α和基质金属蛋白酶9(MMP-9)蛋白的表达,分析HIF-1α介导的高糖微环境对胰腺癌细胞侵袭能力的影响。结果高血糖组患者的肿瘤长径、胆管侵袭率、胰腺癌组织HIF-1α强表达的比例显著高于血糖正常组患者[(2.95±0.73)cm比(2.09±0.70)cm、22.2%比0,44.4%比10.7%],差异均有统计学意义(P值均<0.05)。高血糖组裸鼠胰腺原位移植瘤组织缺氧面积占比[(43.2±2.4)%比(13.5±1.3)%]、移植瘤体积[(1.82±0.88)cm3比(0.75±0.21)cm3]及发生肝转移的比例(5/10比0/10)均显著高于血糖正常组,差异均有统计学意义(P值均<0.05)。对照组、高糖组、缺氧组、高糖+缺氧组BxPC3细胞MMP-9蛋白表达量分别为0.16±0.01、0.47±0.01、0.56±0.01、1.20±0.02,HIF-1Objective To explore the influence and its mechanism of hypoxia inducible factor-1 alpha(HIF-1α)induced hyperglycemia microenvironment on the invasion ability of pancreatic cancer cells.Methods Clinical data of 46 patients who underwent surgical resection of pancreatic cancer in the First Affiliated Hospital of Xi′an Jiaotong University between January 2012 and December 2014 were collected.The patients were divided into euglycemia group(3.9 mmol/L≤blood glucose<6.1 mmol/L)and hyperglycemia group(blood glucose≥6.1 mmol/L)based on the blood glucose level.The tumor size and tumor invasion and metastasis of the two groups were analyzed,and the degree of hypoxia in tumor tissues and the expression of HIF-1αwere detected by immunohistochemistry.Orthotopic pancreatic cancer xenograft diabetes nude mice models were successfully established as the detection of blood glucose≥20 mmol/L continuously for two times with the presence of urine glucose.The nude mice were divided into normal blood glucose group and hyperglycemia group,and tumor size,hypoxia,tumor invasion and metastasis of the orthotopic pancreatic cancer xenograft were observed.Pancreatic cancer BxPC3 cells were cultured in vitro and were divided into high glucose group(cultured in 25 mmol/L glucose medium),hypoxia group(treated by 150μmol/L CoCl2),high glucose+hypoxia group and control group(cultured in routine medium).BxPC3 cells were also transfected with siRNA targeting HIF-1α(siRNA-HIF-1α)with lipofectamine,cultured in routine or high glucose(containing 25 mmol/L glucose)medium,and divided into siRNA-HIF-1αgroup,high glucose+siRNA-HIF-1α,negative control siRNA transfection(siRNA NC)group and high glucose+siRNA NC group.Western blot was used to detect the protein expression levels of HIF-1αand MMP-9 in BxPC3 cells,and the influence of HIF-1αinduced high glucose microenvironment on the invasion of pancreatic cancer cells was analyzed.Results Major diameter of tumor,the rate of bile duct invasion and the percentage of pancreatic cancer specimen

关 键 词：胰腺肿瘤 高血糖症 缺氧 缺氧诱导因子1 Α亚基 

分 类 号：R735.9[医药卫生—肿瘤]