摘要
目的 观察低氧诱导因子-1α（HIF-1α）对胰腺癌细胞生物学行为的影响,并探讨其临床意义.方法 将HIF-1α短发卡RNA（shRNA）重组质粒pGCsi-shHIF-1 α稳定转染人胰腺癌细胞株Panc-1,采用Western blot法检测常氧及缺氧条件下转染细胞HIF-1d的抑制效果,通过细胞噻唑蓝（MTT）增殖实验、划痕实验、侵袭实验及细胞克隆实验,研究HIF-1α对胰腺癌细胞常氧及缺氧状态下黏附力、活动力及增殖力的影响.结果 常氧及缺氧条件下,转染组细胞HIF-1α蛋白表达量均为0,与未转染组（分别为0.78±0.09和1.12±0.12）比较,表达被显著抑制（P＜0.05）.HIF-1α被干扰后,胰腺癌细胞增殖能力明显减弱（P＜0.05）,运动迁移能力显著下降.与未转染的细胞比较,常氧和缺氧下穿过人工基底膜的细胞数明显减少[分别为（18.3±3.8）、（19.5±3.5）个和（99.6±8.9）、（106.6±9.8）个,P＜0.05]；划痕实验显示常氧和缺氧下划痕宽度与未转染组比较明显增宽[分别为（12.22±1.01）、（11.13 ±0.87） mm和（19.21 ±1.06）、（18.23 ±1.34） mm,P＜0.05]；与未转染组比较,单位面积形成克隆球的数目明显减少[分别为（1.89 ±0.16）、（1.98±0.18）个和（1.04±0.11）、（1.13±0.09）个,P＜0.05].结论 RNA干扰可引起Panc-1细胞中HIF-1α的沉默,而HIF-1α表达下调后能有效抑制Panc-1细胞增殖、迁移能力.
Objective To investigate the effect of hypoxia inducible factor-1ot （HIF-1α） on biological behavior of the pancreatic carcinoma cells,and the clinical significance.Methods The recombinant plasmid pGCsi-shHIF-1α was transfected into Panc-1 cells by LipofectamineTM 2000.The inhibitory effect of HIF-1α was measured at protein level by Western blotting under normoxia and hypoxia.In vitro cell adhesion,wound-healing scrape assay,Transwell invasion assay and cell colony formation assay were performed to assess the effect of HIF-1α on the invasive properties of Panc-I cells.Results The expression of HIF-1 α protein in each group of transfected cells was significantly suppressed under normoxia and hypoxia （0.78 ±0.09 and 1.12 ± 0.12,P 〈 0.01）.The migration of the recombinant plasmid-transfected cells was significantly suppressed as compared with that of cells transfected with empty vector （18.3 ± 3.8 and 19.5 ± 3.5 vs.99.6 ± 8.9 and 106.6 ± 9.8,P 〈 0.05）.Colony formation and MTT assays showed that the cell proliferation of the pGCsi-sh HIF-1α transfected cells was decreased as compared with the control groups （1.89 ± 0.16 and 1.98 ± 0.18 vs.1.04±0.11 and1.13±0.09,P〈0.05）.Condusion RNA interference-mediated downregulation of HIF-1α can attenuate the proliferation and migration of Panc-1 cells.
出处
《中华实验外科杂志》
CAS
CSCD
北大核心
2014年第1期32-35,共4页
Chinese Journal of Experimental Surgery
关键词
胰腺肿瘤
RNA干扰
低氧诱导因子-1Α
Pancreatic neoplasms
RNA interference
Hypoxia-inducible factor-1α
