中波紫外線誘導(dǎo)的提前衰老成纖維細胞中環(huán)狀RNA的表達及分析(1)
[摘要]目的：對于中波紫外線誘導(dǎo)的提前衰老的人類真皮成纖維細胞(UVB stress induced premature senescence，UVB-SIPS)，筆者使用基因芯片篩選了其中環(huán)狀RNA的表達譜。方法：構(gòu)建UVB-SIPS模型；采用β-gal染色、細胞周期、CCK8(cell counting kit)檢測衰老成纖維細胞；基因芯片技術(shù)篩選差異表達的環(huán)狀RNA；聚合酶鏈?zhǔn)椒磻?yīng)驗證相關(guān)環(huán)狀RNA；利用Clusterprofiler r package，對上下調(diào)基因分別進行KEGG和GO的富集分析。結(jié)果：與對照組比較(差異表達倍數(shù)≥1.5，P<0.05)，共有472個差異表達的環(huán)狀RNA，其中228個環(huán)狀RNA上調(diào)，244個環(huán)狀RNA下調(diào)。在472個差異表達的環(huán)狀RNA中隨機選擇了8個環(huán)狀RNA進行聚合酶鏈?zhǔn)椒磻?yīng)。其中有5個環(huán)狀RNA(hsa_circRNA_100797，hsa_circRNA_100686，hsa_circRNA_400036，hsa_circRNA_101755，hsa_circRNA_003794)與基因芯片的結(jié)果一致。GO分析顯示，UVB-SIPS組細胞中差異表達的環(huán)狀RNA親本基因與細胞對紫外線輻射的反應(yīng)、DNA修復(fù)復(fù)合物、有絲分裂、微管蛋白結(jié)合等注釋條目有關(guān)；KEGG分析顯示，UVB-SIPS組細胞與未處理組細胞相比，差異表達的環(huán)狀RNA的親本基因可能參與與衰老相關(guān)的細胞周期、p53信號通路、PPAR信號通路等的調(diào)節(jié)。結(jié)論：本研究初步揭示了提前衰老的人類真皮成纖維細胞中環(huán)狀RNA的表達，可為未來研究光老化發(fā)生機制奠定基礎(chǔ)。
, 百拇醫(yī)藥
[關(guān)鍵詞]提前衰老成纖維細胞；環(huán)狀RNA；基因芯片；競爭結(jié)合；生信分析；中波紫外線
[中圖分類號]R329.2 [文獻標(biāo)志碼]A [文章編號]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)
, 百拇醫(yī)藥
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.www.srpcoatings.com(司晨琛 周炳榮 駱丹)
, 百拇醫(yī)藥
[關(guān)鍵詞]提前衰老成纖維細胞；環(huán)狀RNA；基因芯片；競爭結(jié)合；生信分析；中波紫外線
[中圖分類號]R329.2 [文獻標(biāo)志碼]A [文章編號]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)
, 百拇醫(yī)藥
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.www.srpcoatings.com(司晨琛 周炳榮 駱丹)
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