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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150 μmol/L CoCln 2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。n 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(n 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)cmn 3比(0.75±0.21) cmn 3]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(n 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-1α蛋白表达量分别为0.55±0.01、0.85±0.01、0.95±0.01、1.12±0.02，高糖组、缺氧组均显著高于对照组，高糖＋缺氧组又显著高于其他3组，差异均有统计学意义(n P值均<0.05)。siRNA-NC组、高糖＋siRNA-NC组、siRNA-HIF-1α组、高糖＋siRNA-HIF-1α组BxPC3细胞MMP-9蛋白表达量分别为0.30±0.02、0.97±0.00、0.12±0.01、0.18±0.01，HIF-1α蛋白表达量分别为0.98±0.01、2.73±0.04、0.63±0.01、0.91±0.03，siRNA-HIF-1α组均显著低于其他3组，高糖＋siRNA-HIF-1α组均显著低于高糖＋siRNA-NC组，差异有统计学意义(n P值均<0.05)。n 结论:高糖状态下胰腺癌微环境缺氧，通过上调HIF-1α表达促进胰腺癌细胞的侵袭能力。“,”Objective: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-1α were 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 culturedn in vitro and were divided into high glucose group (cultured in 25 mmol/L glucose medium), hypoxia group (treated by 150 μmol/L CoCl n 2), 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-1α group, 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-1α and MMP-9 in BxPC3 cells, and the influence of HIF-1α induced high glucose microenvironment on the invasion of pancreatic cancer cells was analyzed.n Results:Major diameter of tumor, the rate of bile duct invasion and the percentage of pancreatic cancer specimens with strong HIF-1α positivity in hyperglycemia group were significantly higher than those in euglycemia group[(2.95±0.73)cmn vs (2.09±0.70)cm, 22.2%n vs 0, 44.4%n vs10.7%] and all the differences were statistically significant (n P<0.05). Compared with nude mice in the normal blood glucose group, the proportion of hypoxia in orthotopic xenograft tumor tissue [(43.2±2.4)%n vs (13.5±1.3)%] the average volume of orthotopic tumors [(1.82±0.88)cmn 3vs (0.75±0.21)cmn 3] and the rate of liver metastasis (5/10 n vs 0/10) in the hyperglycemia group was greatly higher, and all the difference was statistically significant (all n P values <0.05). The expression of MMP-9 protein in the control group, high glucose group, hypoxia group, and high glucose+ hypoxia group was 0.16±0.01, 0.47±0.01, 0.56±0.01 and 1.20±0.02; HIF-1α protein expression was 0.55±0.01, 0.85±0.01, 0.95±0.01 and 1.12±0.020, respectively. The expression levels of MMP-9 and HIF-1α in high glucose group and hypoxia group were obviously higher than those in control group, and that in high glucose+ hypoxia group were higher than the other three groups, and all the differences were statistically significant (all n P values <0.05). The expression of MMP-9 in siRNA NC group, high glucose + siRNA NC group, siRNA-HIF-1α group, high glucose+ siRNA-HIF-1α group was 0.30±0.02, 0.97±0.00, 0.12±0.01 and 0.18±0.01; HIF -1α protein expression was 0.98±0.01, 2.73±0.04, 0.63±0.01 and 0.91±0.03, respectively. The expression of MMP-9 and HIF-1α in siRNA-HIF-1α group was significantly lower than those in the other three groups (all n P values <0.05); the expression of MMP-9 and HIF-1αin high glucose+ siRNA-HIF-1α group was significantly lower than that of the high glucose+ siRNA-NC group, and all the difference was statistically significant (all n P values <0.05).n Conclusions:High glucose state promotes the hypoxia in the microenvironment of pancreatic cancer and the invasion ability of pancreatic cancer cells by up-regulating the expression of HIF-1α.