中波紫外线诱导的提前衰老成纤维细胞中环状RNA的表达及分析(1)
[摘要]目的：對于中波紫外线诱导的提前衰老的人类真皮成纤维细胞(UVB stress induced premature senescence，UVB-SIPS)，笔者使用基因芯片筛选了其中环状RNA的表达谱。方法：构建UVB-SIPS模型；采用β-gal染色、细胞周期、CCK8(cell counting kit)检测衰老成纤维细胞；基因芯片技术筛选差异表达的环状RNA；聚合酶链式反应验证相关环状RNA；利用Clusterprofiler r package，对上下调基因分别进行KEGG和GO的富集分析。结果：与对照组比较(差异表达倍数≥1.5，P<0.05)，共有472个差异表达的环状RNA，其中228个环状RNA上调，244个环状RNA下调。在472个差异表达的环状RNA中随机选择了8个环状RNA进行聚合酶链式反应。其中有5个环状RNA(hsa_circRNA_100797，hsa_circRNA_100686，hsa_circRNA_400036，hsa_circRNA_101755，hsa_circRNA_003794)与基因芯片的结果一致。GO分析显示，UVB-SIPS组细胞中差异表达的环状RNA亲本基因与细胞对紫外线辐射的反应、DNA修复复合物、有丝分裂、微管蛋白结合等注释条目有关；KEGG分析显示，UVB-SIPS组细胞与未处理组细胞相比，差异表达的环状RNA的亲本基因可能参与与衰老相关的细胞周期、p53信号通路、PPAR信号通路等的调节。结论：本研究初步揭示了提前衰老的人类真皮成纤维细胞中环状RNA的表达，可为未来研究光老化发生机制奠定基础。
, 百拇医药
[关键词]提前衰老成纤维细胞；环状RNA；基因芯片；竞争结合；生信分析；中波紫外线
[中图分类号]R329.2 [文献标志码]A [文章编号]1008-6455(2018)12-0128-04
The Expression and Analysis of Circular RNA in Fibroblasts Induced by Ultraviolet B
SI Chen-chen，ZHOU Bing-rong，LUO Dan
(Department of Dermatology， the First Affiliated Hospital of Nanjing Medical University，Nanjing 210029，Jiangsu，China)
, 百拇医药
Abstract： Objective CircRNAs expression profiling was screened in human fibroblasts (HFs) with gene chip in our study. Methods The senescence of cells was verified by β-gal staining， cell cycle and CCK8 (cell counting kit). The differently expressed circRNA ware screened by gene chip.Some circRNAs were tested by qRT-PCR (real-time quantitative polymerase chain reaction) randomly. The Clusterprofiler r package was used to analyse GO and KEGG pathway of up-regulated and down-regulated circRNAs. Results Compared to the control group (fold change ≥1.5，P<0.05)， a total of 472 circRNAs in the UVB-SIPS group were differently expressed. Among them， 228 circRNAs were up-regulated and the other 244 down-regulated. The microarrays of eight circRNAs selected from the 472 differently expressed circRNAs were retested by qRT-PCR. The results of five (hsa_circRNA_100797， hsa_circRNA_100686， hsa_circRNA_400036， hsa_circRNA_101755， hsa_circRNA_003794) were in accordance with their chip results. Go analysis showed that the parent genes of differently expressed circRNA in UVB-SIPS group were related to the annotataion of cellular response to UV， DNA repair complex， mitotic nuclear division， tubulin protein binding and so on. KEGG analysis implicated the parent genes of differently expressed circRNA in UVB-SIPS group may be involved in regulating the pathway of cell cycle， p53 signal pathway， PPAR signal pathway when compared to blank group. Conclusion This study uncovered the profiling of circRNA in premature senescent human fibroblasts for the first time and lay the foundation for studying the mechanism of photoaging., http://www.100md.com(司晨琛 周炳荣 骆丹)
, 百拇医药
[关键词]提前衰老成纤维细胞；环状RNA；基因芯片；竞争结合；生信分析；中波紫外线
[中图分类号]R329.2 [文献标志码]A [文章编号]1008-6455(2018)12-0128-04
The Expression and Analysis of Circular RNA in Fibroblasts Induced by Ultraviolet B
SI Chen-chen，ZHOU Bing-rong，LUO Dan
(Department of Dermatology， the First Affiliated Hospital of Nanjing Medical University，Nanjing 210029，Jiangsu，China)
, 百拇医药
Abstract： Objective CircRNAs expression profiling was screened in human fibroblasts (HFs) with gene chip in our study. Methods The senescence of cells was verified by β-gal staining， cell cycle and CCK8 (cell counting kit). The differently expressed circRNA ware screened by gene chip.Some circRNAs were tested by qRT-PCR (real-time quantitative polymerase chain reaction) randomly. The Clusterprofiler r package was used to analyse GO and KEGG pathway of up-regulated and down-regulated circRNAs. Results Compared to the control group (fold change ≥1.5，P<0.05)， a total of 472 circRNAs in the UVB-SIPS group were differently expressed. Among them， 228 circRNAs were up-regulated and the other 244 down-regulated. The microarrays of eight circRNAs selected from the 472 differently expressed circRNAs were retested by qRT-PCR. The results of five (hsa_circRNA_100797， hsa_circRNA_100686， hsa_circRNA_400036， hsa_circRNA_101755， hsa_circRNA_003794) were in accordance with their chip results. Go analysis showed that the parent genes of differently expressed circRNA in UVB-SIPS group were related to the annotataion of cellular response to UV， DNA repair complex， mitotic nuclear division， tubulin protein binding and so on. KEGG analysis implicated the parent genes of differently expressed circRNA in UVB-SIPS group may be involved in regulating the pathway of cell cycle， p53 signal pathway， PPAR signal pathway when compared to blank group. Conclusion This study uncovered the profiling of circRNA in premature senescent human fibroblasts for the first time and lay the foundation for studying the mechanism of photoaging., http://www.100md.com(司晨琛 周炳荣 骆丹)
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