p53-repressed miRNAs are Involved with E2F in a Feed Forward Loop Promoting Proliferation

miRNA expression profiles of WI38 primary human fibroblasts with an active or inactive p53. Cells were compared under normal untreated conditions (young and proliferating cells), after DNA damage with Doxorubicin, and upon entry into replicative senescence. Keywords: miRNA, WI-38, p53, GSE56, Senescence, Doxorubicin, Cancer, DNA-damage, fibroblasts. 6 samples of WI38 cells were analyzed on 12 Exiqon miRcurry LNA arrays in biological duplicates (2 different cell culture plates for each experimental condition). The six conditions included: 1. [Con_Young] - Primary Young WI38 cells (passage 20) with a control retroviral vector (pLXSN-NEO). Untreated. 2. [GSE_Young] -Primary Young WI38 cells (passage 20) with a retroviral vector encoding for the p53-inactivating peptide GSE56 (pLXSN-NEO-GSE56).Untreated. 3. [Con_Dox] -Primary Young WI38 cells (passage 20) with a control retroviral vector (pLXSN-NEO). Treated with Doxorubicin (0.2 micrograms/ml) for 24 hours). 4. [GSE_Dox] Primary Young WI38 cells (passage 20) with a retroviral vector encoding for the p53-inactivating peptide GSE56 (pLXSN-NEO-GSE56). Treated with Doxorubicin (0.2 micrograms/ml) for 24 hours). 5. [Con_Old] - Sesescent WI38 cells (passage 30) with a control retroviral vector (pLXSN-NEO). Untreated. 6. [GSE_Old] - Senescent WI38 cells (passage 26) with a retroviral vector encoding for the p53-inactivating peptide GSE56 (pLXSN-NEO-GSE56).Untreated. RNA was extracted with TRI-Reagent and sent for labeling and hybridization in Exiqon laboratories (In Denamark). Samples were labeled with Cy5. Reference sample (Cy3) was an RNA mix of all samples. Log2 for Ratio(Cy5/Cy3) was used for further analysis.

本数据集为携带活性或失活p53的人胚肺成纤维细胞WI38的microRNA（miRNA）表达谱。本研究对比了三类细胞状态：正常未处理的年轻增殖细胞、经阿霉素（Doxorubicin）处理造成DNA损伤的细胞，以及进入复制性衰老的细胞。
关键词：miRNA、WI-38、p53、GSE56、复制性衰老、阿霉素、癌症、DNA损伤、成纤维细胞。
共对6组WI38细胞样本进行检测，采用12张艾克森（Exiqon）miRCURY LNA芯片完成实验，每组实验条件设置生物学重复（即每个实验组使用2块不同的细胞培养板）。6组实验条件具体如下：
1. [Con_Young]：携带对照逆转录病毒载体（pLXSN-NEO）的年轻初代WI38细胞（传代20次），未接受任何处理。
2. [GSE_Young]：携带编码p53失活肽GSE56的逆转录病毒载体（pLXSN-NEO-GSE56）的年轻初代WI38细胞（传代20次），未接受任何处理。
3. [Con_Dox]：携带对照逆转录病毒载体（pLXSN-NEO）的年轻初代WI38细胞（传代20次），经0.2μg/mL阿霉素处理24小时。
4. [GSE_Dox]：携带编码p53失活肽GSE56的逆转录病毒载体（pLXSN-NEO-GSE56）的年轻初代WI38细胞（传代20次），经0.2μg/mL阿霉素处理24小时。
5. [Con_Old]：携带对照逆转录病毒载体（pLXSN-NEO）的衰老WI38细胞（传代30次），未接受任何处理。
6. [GSE_Old]：携带编码p53失活肽GSE56的逆转录病毒载体（pLXSN-NEO-GSE56）的衰老WI38细胞（传代26次），未接受任何处理。
总RNA采用TRI-Reagent试剂提取，随后送至丹麦艾克森实验室完成标记与杂交实验。所有样本均采用Cy5荧光标记，以全部样本混合的RNA作为参考样本（Cy3标记），后续分析采用Cy5/Cy3比值的以2为底的对数值（log₂(Cy5/Cy3)）。