胰岛素对人胰腺癌PANC1细胞增殖与侵袭能力的影响

摘要

目的 研究胰岛素对人胰腺癌PANC1细胞增殖、侵袭能力及细胞低氧诱导因子(HIF-1α)、血管内皮生长因子( VEGF)表达的影响.方法 采用0.1、1、10、100 nmol/L胰岛素处理PANC1.噻唑蓝法和Transwell小室侵袭实验检测细胞增殖和侵袭能力；蛋白质印迹法和实时PCR法检测增殖性细胞核抗原(PCNA)及HIF-1α、VEGF蛋白和mRNA的表达.结果 胰岛素呈剂量依赖性诱导PANC1细胞的增殖,上调HIF-1α、VEGF蛋白表达.100 nmol/L胰岛素干预4d,PANC1细胞的PCNA蛋白表达显著上调(1.196±0.014比1.157±0.013,P＜0.05)；Transwell小室穿膜细胞数量显著增加[(141.0±2.1)个比(89.0±1.4)个,P＜0.05]；HIF-1α蛋白表达显著上调(1.139±0.020比0.598 ±0.013,P＜0.05),但HIF-1α mRNA表达无明显变化；VEGF蛋白表达显著上调(1.011±0.023比0.627±0.013,P ＞0.05),VEGF mRNA表达亦显著上调(0.970±0.016比0.350±0.013,P＜0.05).结论 高胰岛素微环境可诱导PANC1细胞增殖,并通过上调HIF-1α及VEGF的表达增强细胞的侵袭能力.%Objective To investigate the effects of insulin on the proliferation and invasion of human pancreatic cancer cells PANC1,and on its HIF-1α,VEGF expression.Methods PANC1 was pretreated with insulin of different concentrations (0.1,1,10,100 nmol/L).The proliferation of PANC1 was tested by MTTmethod,and transwell assay was used to test the invasion ability of PANC1.HIF-1α,VEGF and PCNA protein expression was assessed by Western blots,and HIF-1α,VEGF mRNA was detected by real-time PCR.Results Insulin could increase the proliferation of PANC1 in a dose-dependent manner (p ＜0.05 ),and increase the expression of HIF-1α,VEGF protein.After 100 nmol/L insulin treatment for4 d,the PCNA protein expression in the insulin group was significantly higher than that in the control group (1.196 ±0.014 vs 1.157 ±0.013,P ＜ 0.05).The cancer cells passed through the chamber in insulin group were much more than that in the control group ( 141.0 ± 2.1 vs 89.0 ± 1.4,P ＜0.05 ).The expression of HIF-1α protein was significantly increased (1.139 ±0.020 vs 0.598 ±0.013,P ＜0.05),while there was no significant change of HIF-1αmRNA expression.Both the expression of VEGF protein and mRNA were significantly increased (1.011 ± 0.023 vs 0.627 ± 0.013 0.970 ± 0.016 vs 0.350 ± 0.01 3,P ＜ 0.05 ).Conclusions High insulin microenvironment could enhance the proliferation and invasion of PANC1 cells by up-regulating the expression of HIF-1α and VEGF.