He Guodong, Jiang Yi, Zhang Bo, Wu Guohao
Department of General Surgery, Institute of General Surgery, 180 Fenglin Road, Zhongshan Hospital, Fudan University, Shanghai 200032, China.
Asia Pac J Clin Nutr. 2014;23(1):174-80. doi: 10.6133/apjcn.2014.23.1.14.
The aim of this study is to explore the possible role of HIF-1α in glucose metabolism, proliferation and apoptosis of pancreatic cancerous cells.
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.
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.
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.
本研究旨在探讨缺氧诱导因子-1α（HIF-1α）在胰腺癌细胞糖代谢、增殖及凋亡过程中可能发挥的作用。
将胰腺癌BxPC-3细胞分别在常氧及低氧（3% O₂）条件下培养。采用MTT法检测细胞增殖情况，用膜联蛋白V/碘化丙啶（Annexin V/PI）染色法检测细胞凋亡情况。通过蛋白质免疫印迹法（Western-blot）和实时荧光定量聚合酶链反应（Realtime PCR）检测丙酮酸脱氢酶激酶（PDK1）、乳酸脱氢酶（LDHA）、丙酮酸激酶M2（PKM2）及柠檬酸合酶（CS）的表达。
低氧条件下，HIF-1α表达及乳酸生成增加。与有氧条件相比，糖代谢酶PDK1、LDHA、PKM2的表达也增加。低氧处理对CS的表达影响较小。低氧条件下，敲低HIF-1α可抑制乳酸生成及PDK1、LDHA和PKM2的表达。敲低HIF-1α可抑制胰腺癌BxPC-3细胞在低氧条件下的生长并诱导其凋亡。
低氧条件下，HIF-1α表达上调，可能通过上调糖酵解相关酶的表达，导致BxPC-3细胞糖酵解增加。体外实验中，敲低HIF-1α可抑制胰腺癌BxPC-3细胞的增殖并促进其凋亡。
