摘要
目的:研究低氧对体外培养胃上皮细胞株GES-1中MICA基因及蛋白表达的影响。方法:GES-1细胞常规细胞培养,用含10%胎牛血清的高糖DMEM培养液,加青霉素100U/mL,链霉素100mg/L。培养细胞于生长的对数期,将细胞(5×105/mL)置于Jouan三气培养箱内,建立GES-1细胞体外低氧培养模型,设置缺氧条件为37℃,5%CO2,5%O2,90%N2,在缺氧处理后0、12、24和72h,半定量逆转录聚合酶链反应法和免疫组织化学技术分别检测MICA基因和蛋白的表达变化。结果:在低氧条件下培养0、12、24和72h后MICA mRNA/β-actin吸光度比值和MICA蛋白的表达吸光度值在不同刺激时间其表达的差别有统计学意义(均P<0.01)。Pearson积差相关分析显示MICA mRNA和蛋白水平表达呈正相关。结论:低氧可以增加胃上皮细胞株GES-1中MICA基因的表达。
Objective：To investigate the effect of hypoxia on MICA expression in gastric epithelial cell strain-1（GES-1） and the mechanism thereof.Methods：Hypoxia culture model（37 ℃,5% CO2,5% O2,90% N2）in vitro was established to test the expression of MICA in GES-1 cell line.After cultured in hypoxia circumstance for a given period of time 0 hour（group A）,12 hours（group B）,24 hours（group C） and 72 hours（group D） MICA expression was tested in mRNA level and protein level by SQ-RT-PCR and immunocytochemistry analysis.Results：The expression of MICA increased after treatment of hypoxia.The expression of MICA protein increased equally with mRNA confirmed by immunocytochemistry and RT-PCR,and the positive correlation was found by Pearson product-moment correlation analysis.Conclusion：MICA gene and protein can be induced remarkably by hypoxia in GES-1.
出处
《天津医药》
CAS
北大核心
2010年第7期599-601,共3页
Tianjin Medical Journal
基金
青海省科技厅重点科技攻关项目(项目编号:2006-N-175)
青海大学医学院中青年科技基金(项目编号:qy200807)
关键词
细胞低氧RNA
信使
胃
上皮细胞
细胞系
基因表达
逆转录聚合酶链反应
免疫组织化学
cell hypoxia RNA
messenger stomach epithelial cells cell line gene expression reverse transcriptase polymerase chain reaction immunohistochemistry
