摘要
目的应用基因本体(GO)数据库分析中波紫外线(UVB)对人永生化角质形成细胞(Ha Ca T细胞)micro RNA(mi RNA)表达的影响。方法 Ha Ca T细胞根据UVB强度(100m J/cm^2、200 mJ/cm^2、400 mJ/cm^2)及辐照时间(12～24 h)的不同组合,完成7张mi RNA芯片[(对照、100 mJ/cm^2(12 h)、100 mJ/cm^2(24 h)、 200 mJ/cm^2(12 h)、200 mJ/cm^2(24 h)、400 mJ/cm^2(12 h)、400 mJ/cm^2(24 h)]的制作及扫描,筛选差异表达的mi RNAs,采用Target Scan进行靶基因预测,并对其进行基于GO数据库的功能富集分析。结果不同强度UVB辐照HaCaT细胞后均出现差异miRNA表达,其中400 mJ/cm^2 UVB处理组的miRNA差异表达最为明显,根据差异倍数(fc)筛选9个miRNAs拟进行后续实验研究,分别为上调表达的miR-8063、miR-1273f、miR-4497、miR-4778-5p、miR-6510-5p和下调表达的miR-1973、miR-5100、miR-205-3p、miR-4485-3p。靶基因预测及GO富集分析发现部分靶基因功能涉及T淋巴细胞介导的细胞毒作用、T淋巴细胞耐受、B淋巴细胞分化、自然杀伤细胞增生、树突细胞抗原处理提呈、免疫球蛋白产生以及中性粒细胞稳态等方面。结论 UVB作用HaCaT细胞后存在差异表达miRNAs,其靶基因功能与体液免疫和细胞免疫相关。
Objective To investigate the microRNA(miRNA) expression profiles in HaCaT cells induced by UVB based on gene ontology. Methods Differential expression of miRNAs were screened by preparing and scanning 7 miRNA chips of control, 100 mJ/cm^2-12 h, 100 mJ/cm^2-24 h, 200 mJ/cm^2-12 h, 200 mJ/cm^2-24 h, 400 mJ/cm^2-12 h and 400 mJ/cm^2-24 h group according to different combination of intensity and duration of UVB. Target genes were predicted by TargetScan and enrichment analysis was performed based on gene ontology. Results To be compared with the control group, HaCaT cells induced by UVB of different intensity showed differential expressions of miRNAs, which of 400 mJ/cm^2 group were the most significant. Nine miRNAs, including 5 up-regulated: miR-8063, miR-1273 f, miR-4497, miR-4778-5p and miR-6510-5p as well as 4 down-regulated: miR-1973, miR-5100, miR-205-3p and miR-4485-3p, were selected based on fold change(fc) for the following study. Prediction and enrichment analysis on target genes for miRNAs of HaCaT cells of 400 mJ/cm^2-UVB group showed several gene functions including positive regulation of T cell mediated cytotoxicity, positive regulation of T cell tolerance induction, immature B cell differentiation, natural killer cell proliferation, dendritic cell antigen processing and presentation, negative regulation of immunoglobulin production and neutrophil homeostasis, etc. Conclusion HaCaT cells induced by UVB showed differential expressions of miRNAs, the functions of the target genes may be related to humoral and cellular immunity.
作者
杨玲
许速
胡中华
YANG Ling;XU Su;HU Zhong-hua(Department of Dermatology,Kunming General Hospital of PLA,Kunming 650032,China)
出处
《实用皮肤病学杂志》
2018年第6期329-332,336,共5页
Journal of Practical Dermatology
基金
国家自然科学基金资助项目(81502743)
