硫化镍转化人支气管上皮细胞基因差异表达芯片的研究

Study on gene expression pattern in 16HBE cells treated with NiS

背景与目的近年研究发现硫化镍(NiS)可引起人支气管上皮细胞(humanbronchialepithelialcell,16HBE)发生恶性转化及转化细胞致癌性,其机制可能与NiS引起基因突变、多种转录因子的异常表达有关。为此,本研究利用NiS恶性转化的体外培养细胞模型,用cDNA微阵列技术研究经NiS转化后的16HBE细胞基因的差异表达,从基因组水平探讨NiS所致细胞恶变的相关基因差异改变。方法分别提取16HBE和经NiS单独处理发生转化的NiS16HBE细胞的总RNA,逆转录合成cDNA并以Cy3dCTP和Cy5dCTP荧光素分别标记制作探针。探针混合后与含4000个人类基因的芯片杂交。以ScanArray4000扫描仪扫描芯片,以GenPixPro3.0软件分析荧光信号,然后对差异表达的基因进行生物学信息分析。结果NiS16HBE细胞样本与16HBE细胞样本比较,呈现差异表达的基因有151个(3.78%),其中81个(53.64%)上调,70个(46.36%)下调。结论NiS的转化作用与应激反应基因、免疫相关基因、DNA合成和修复基因、代谢相关基因、原癌基因和抑癌基因等多类基因的异常表达有关。

Background and objective Recent researches have found that NiS can cause the malignant transforming activity and carcinogenicity on human bronchial epithelial cells （16HBE）. Its molecular mechanism may be involved in mutation of genes and abnormal expression of transcription factors on 16HBE. And so, this study takes advantage of a model of 16HBE transformed by NiS and screens the differentially expressed genes between 16HBE cells and NiS treated 16HBE cells （NiS-16HBE） using cDNA microarray. Methods The total RNA was extracted from 16HBE cells and NiS-16HBE cells. The cDNA probes were prepared by labeling with Cy3-dCTP and Cy5-dCTP respectively through reverse transcription. The mixed probes were then hybridized to the cDNA microarray chips containing 4000 human genes. The chips were scanned by ScanArray 4000 laser scanner. The acquired fluorescent signals were analyzed by GenPix Pro 3.0 software. Bioinformation function of those differentially expressed genes was analysed. Results A total of 151 genes exhibited differential expression between 16HBE cells and NiS-16HBE cells. The expression of 70 genes （46.36%） was down-regulated and that of 81 genes （53. 640/00） was up regulated. Conclusion The regulation of genes including stress response genes, immune related genes, DNA synthesis and repair genes, metabolism genes, pro-oncogenes and tumor suppressor genes may be involved in transforming activity of NiS.