摘要
目的:探讨缺氧应激对HepG-2细胞甲胎蛋白(alpha fetoprotein,AFP)和细胞间粘附分子(intercellular adhe-sion molecule,ICAM)-1表达的影响。方法:缺氧(5%O2、5%CO2、90%N2)和常氧培养HepG-2细胞。细胞侵袭实验测定HepG-2细胞侵袭能力;四甲基偶氮唑蓝(MTT)比色法测定细胞增殖活性;逆转录聚合酶链反应技术测定常氧组和缺氧12、24h组HepG-2细胞AFPmRNA和ICAM-1mRNA的表达量。流式细胞仪技术测定ICAM-1蛋白表达,放射免疫法测定HepG-2细胞培养上清AFP蛋白表达。结果随着缺氧时间的延长HepG-2细胞的侵袭能力、增殖活性、AFP、ICAM-1 mRNA和蛋白表达量明显增高(P<0.01)。结论:缺氧应激下人HepG-2细胞侵袭能力增强的机制之一是ICAM-1基因表达上调;HepG-2细胞增殖活性增强的机制之一是HepG-2细胞AFP基因的表达上调。
[Objective] To investigate whether hypoxia stress regulates expressions of AFP and ICAM-1 in HepG-2 cell.[Method] HepG-2 cells were cultured under hypoxia(5%O2,5%CO2 and 90%N2) for 12,24 h or normoxia.The proliferation in HepG-2 cell was detected by methyl thiazolyl tetrazolium(MTT).Cell invasion and migration assay was used to decide the invasion in HepG-2 cell.The expressions of AFP and ICAM-1 mRNA in HepG-2 cell were determined by reverse transcription polymerase chain reaction(RT-PCR).The expression of ICAM-1 protein in HepG-2 cell was studied by flow cytometry assay.The expression of AFP protein in HepG-2 cell was determined by radio-immunity assay(RIA).[Result] With the hypoxia time extending,the expressions of AFP and ICAM-1 gene were dramatically enhanced(P<0.01).[Conclusion] Up-regulation of AFP and ICAM-1 contributes to the enhancement of proliferation,invasion in HepG-2 cell by hypoxia stress respectively.
出处
《浙江医学教育》
2012年第1期44-47,共4页
Zhejiang Medical Education
基金
青海省卫生厅2006年指导性科研项目(编号:200601)
