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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。
Objective To investigate the changes of gene expression of bronchial epithelial cells after tumor necrosis factor (TNF) -α activation by cDNA microarray. Methods The total RNA of human bronchial epithelial cell line (BEAS-2B) was extracted. The gene expression of BEAS-2B cells cultured in normal and TNF-α was detected by microarray. The gene expression of BEAS-2B was detected by reverse transcription polymerase chain reaction PCR) confirmed that the concentration of cytokines in TNF-α-activated BEAS-2B cell culture fluid was detected by flow cytometry (CBA). Results ICAM-1, IL-8, MCP-1, TNF-α, IL-6 gene expression in BEAS-2B cells after TNF-α activation (Up-regulated by 6.5, 8.2, 1.4 and 9.5, respectively) and the up-regulated genes were confirmed by RT-PCR. Secretion of cytokines in cultured BEAS-2B cells after TNF-α activation (BEAS-2B cells were cultured without TNF-α and the secretion of IL-6, IL-8 and MCP- · 83 ± 18 · 20 ng / L and (1771 · 33 ± 312 · 67) ng / L, P <0.001 · (277 · 97 ± 50 · 76) ng / L and (7579 · 20 ± 797 · 15) ng / L, P <0.001, (741.53 ± 129.91) ng / L and (12228.557 ± 1897.58) ng / L, P <0.001) Consistent. Conclusion The screening of activated bronchial epithelial cell gene by gene chip method is of great significance in studying the role of bronchial epithelium in airway diseases.