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  • Title: The effect of HIF-1α on glucose metabolism, growth and apoptosis of pancreatic cancerous cells.
    Author: He G, Jiang Y, Zhang B, Wu G.
    Journal: Asia Pac J Clin Nutr; 2014; 23(1):174-80. PubMed ID: 24561986.
    Abstract:
    OBJECTIVES: The aim of this study is to explore the possible role of HIF-1α in glucose metabolism, proliferation and apoptosis of pancreatic cancerous cells. METHOD: The pancreatic cancerous BxPC-3 cells were cultured in normoxia or hypoxia (3% O2), respectively. Cell proliferation was determined by MTT assay, apoptosis was determined by Annexin V/PI staining. Expression of Pyruvate dehydrogenase kinase (PDK1), Lactate dehydrogenase (LDHA), pyruvate kinase M2 (PKM2) and citrate synthase (CS) was determined by Western-blot and Realtime PCR. RESULTS: Under hypoxia, the expression of HIF-1α and the lactate production were increased. The expression of glucose metabolic enzymes PDK1, LDHA, PKM2 was also increased compared with that under aerobic condition. Hypoxia treatment had little effect on expression of CS. Under hypoxia, knockdown of HIF-1α inhibited the production of lactate and the expression of PDK1, LDHA and PKM2. Knockdown of HIF-1α repressed the growth of pancreatic cancer BxPC-3 cells and induced apoptosis of the cells under hypoxia. CONCLUSION: Under hypoxia, the expression of HIF-1α is induced, leading to the increase of glycolysis in BxPC-3 cells possibly through upregulation of the enzymes related to glycolysis. HIF-1α knockdown can inhibit the prolife ratio and promote apoptosis of pancreatic cancerous BxPC-3 cells in vitro. 目的：探讨HIF-1α 基因沉默对胰腺癌细胞生长和凋亡的影响；探讨HIF－1α 对胰腺癌细胞糖代谢的调控作用。方法：胰腺癌细胞分别在有氧及缺氧 (3%O2)条件下培养。利用MTT 实验绘制胰腺癌细胞生长曲线，流式细胞仪检 测细胞凋亡；利用乳酸试剂盒测定乳酸含量；利用定量PCR 和Western Blot 方法测定丙酮酸脱氢酶激酶1(PDK1)、乳酸脱氢酶A(LDHA)、丙酮酸激酶 M2(PKM2)和柠檬酸合成酶(CS)的表达，观察HIF-1α 沉默前后相关检测指标 的变化。结果：缺氧条件下，HIF-1α mRNA 和蛋白的表达均明显升高；乳酸 的生成、糖代谢酶PDK1、LDHA、PKM2 mRNA 和蛋白的表达均明显升高， 同有氧条件下比较，差别有统计学意义；CS mRNA 和蛋白的表达同有氧条件 下比较，差别无统计学意义。缺氧条件下，HIF-1α 基因沉默后，乳酸的生 成、糖代谢酶PDK1、LDHA、PKM2 mRNA 和蛋白的表达均受到明显抑制， 与空载体组比较，差别有统计学意义；CS mRNA 和蛋白的表达没有受到明显 影响，与空载体组比较，差别无统计学意义。缺氧条件下，HIF-1α 基因沉默 后，胰腺癌BxPC-3 细胞的生长受到抑制，凋亡率升高，同空载体组比较，差 别有统计学意义。结论：在缺氧条件下，HIF-1α 的表达明显增强，它可能通 过上调其下游糖酵解相关酶的表达，增强胰腺癌细胞的糖酵解代谢；HIF-1α 沉默在体外能抑制胰腺癌BxPC-3 细胞的生长，使其凋亡率升高。
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