【摘要】 目的探讨基因芯片筛选肿瘤坏死因子(TNF)-α活化后支气管上皮细胞基因表达的变化。方法提取支气管上皮细胞系(BEAS-2B细胞)总RNA,用基因芯片检测正常培养和经TNF-α活化的BEAS-2B细胞基因表达,对上调表达的基因用逆转录聚合酶链反应(RT-PCR)确证,用流式细胞微珠方法(CBA)检测TNF-α活化的BEAS-2B细胞培养液中相关细胞因子浓度。结果TNF-α活化后BEAS-2B细胞中细胞间黏附分子(ICAM)-1、白细胞介素(IL)-8、单核细胞趋化蛋白(MCP)-1、TNF-α、IL-6基因表达显著上调(分别上调6·5、8·2、3·1、4·9和3·5倍),表达上调的基因用RT-PCR确证结果基本一致。TNF-α活化后BEAS-2B细胞培养液中细胞因子的分泌(BEAS-2B细胞培养过程中有、无TNF-α存在,IL-6、IL-8和MCP-1的分泌分别为:(117·83±18·20)ng/L和(1771·33±312·67)ng/L,P<0·001;(277·97±50·76)ng/L和(7579·20±797·15)ng/L,P<0·001;(741·53±129·91)ng/L和(12228·57±1897·58)ng/L,P<0·001),与相应基因的表达一致。结论用基因芯片方法筛选活化后支气管上皮细胞基因表达对研究支气管上皮在气道性疾病中的作用具有重要意义。更多还原

【Abstract】 Objective To study the gene expression in bronchial epithelial cells (BEAS-2B) activated by TNF-α.Methods Total RNA was extracted from BEAS-2B cells and gene expression was screened in BEAS-2B cells treated with or without TNF-α using cDNA array. The gene expression was confirmed by reverse transcription polymerase chain reaction (RT-PCR), and cytokines in supernatant of BEAS-2B cells culture were assayed by cytometric beads array (CBA) kit in flow cytometry.Results Gene expression of ICAM-1、interleukin (IL)-8、monocyte chemotactic protein (MCP)-1、tumor necrosis factor (TNF)-α and IL-6 were upregulated in BEAS-2B cells activated by TNF-α and the results were confirmed by RT-PCR. Cytokine concentration in culture supernatant of BEAS-2B cells activated by TNF-α correlated with the gene expression.Conclusion Study of gene expression in activated bronchial epithelial cells contributes to the understanding of the immunopathology in airway diseases.更多还原