摘要
目的:研究α干扰素(IFN-α-1b)对人肝癌细胞系HepG2细胞MMP-2和TIMP-2表达水平的影响,探讨IFN-α抗肝细胞癌生长与转移复发的相关分子机制。方法:培养HepG2细胞,在不同剂量的IFN-α干预下,于不同时间提取上清液,通过ELISA方法检测MMP-2、TIMP-2蛋白的表达浓度。逆转录聚合酶链反应(RT-PCR)方法半定量检测IFN-α干预组与对照组MMP-2、TIMP-2基因mRNA的表达。结果:在培养HepG2细胞24及48h后,IFN-α剂量为3 000IU/mL时的MMP-2蛋白浓度分别为(1 725.3±300.7)及(2 899.8±343.2)ρg/mL,与空白对照组相比明显降低,差异有统计学意义,P值分别为0.001和0.000;各时间段TIMP-2浓度与对照组比较差异均无统计学意义;MMP-2、TIMP-2浓度比值较对照组显著降低,差异有统计意义,P值分别为0.003和0.001。48h干预组(3 000IU/mL)与对照组比较MMP-2mRNA表达水平差异有统计学意义,F=2.711,P=0.005。结论:大剂量IFN-α可通过抑制HepG2细胞MMP-2的表达水平下调MMP-2、TIMP-2浓度比值,并呈剂量效应关系。IFN-α对HepG2细胞MMP-2表达的抑制作用在其抗肝细胞癌生长与转移复发中发挥一定作用。
OBJECTIVE： To investigate the effects of IFN-α on the expression of MMP-2/TIMP-2 and the ratio of MMP-2/TIMP-2 in human hepatocellular carcinoma（HCC） cell line HepG2 cells,and explore its molecular mechanism of inhibiting the growth and metastasis of HCC.METHODS： MMP-2/TIMP-2 concentration were measured in the supernate of HepG2 cells incubated in DMEM or exposure to differing dosages of IFN-α by ELISA.The expression of MMP-2 and TIMP-2 mRNA were semiquantitatively detected by reverse transcription polymerase chain reaction（RT-PCR）.RESULTS： MMP-2 concentration were（1 725.3±300.7） and（2 899.8±343.2） ρg/mL in the supernate of HepG2 cells incubated for 24 and 48 h,exposure to IFN-α 3 000 IU/mL.There were significant differences compared with the controls（P=0.001 and P=0.000）,and no significant differences were found between TIMP-2 concentration of intervention groups and the control groups.But significant differences were found between MMP-2/TIMP-2 ratio after incubation for 24 or 48 h（P=0.003 and P=0.001）.Furthermore,there was significant difference of MMP-2 mRNA levels between 48 h group（3 000 IU/mL） and the controls（F=2.711,P=0.005）.CONCLUSIONS： IFN-α treatment significantly lower MMP-2/TIMP-2 ratio by inhibiting the expression of MMP-2 in HepG2 cells in a dose-dependent manner.The down-regulation of MMP-2 levels contributes,at least in part,to its activity of inhibiting the growth and metastasis of HCC.
出处
《中华肿瘤防治杂志》
CAS
北大核心
2012年第3期201-204,共4页
Chinese Journal of Cancer Prevention and Treatment
