The effect of SNAIL transcription factor on microRNA transcriptome in rhabdomyosarcoma

The main goal of the project was to analyze the effect of SNAIL transcription factor on microRNA expression profile in rhabdomyosarcoma (RMS) cells using the next generation sequencing. Differential expression of microRNAs between three groups was compared in RH30 alveolar RMS cells: WT (WT), shCTRL (modified with control shRNA vector) and shSNAIL (modified with shRNA against SNAIL). Different groups were compared to investigate the effect of SNAIL silencing on microRNA up- or downregulation. Overall design: Total RNA from three different cell types (RH30 WT, shCTRL, shSNAIL) was isolated at three independent time points (three independent experiments) and microRNA expression profile was analyzed by the next generation seqeuncing at Exiqon Services, Denmark.

本项目的核心研究目标为借助下一代测序（next generation sequencing）技术，解析SNAIL转录因子对横纹肌肉瘤（rhabdomyosarcoma, RMS）细胞中microRNA表达谱的调控效应。实验以RH30肺泡横纹肌肉瘤细胞为研究对象，设置三组实验组别：WT（野生型）组、shCTRL（对照短发卡RNA载体转染组）以及shSNAIL（靶向SNAIL的短发卡RNA转染组），通过比较三组间microRNA的差异表达水平，探究SNAIL基因沉默对microRNA上调与下调表达的影响。实验整体设计如下：分别从RH30 WT、shCTRL、shSNAIL三种细胞类型中提取总RNA，每个组别均设置三次独立生物学重复（三次独立实验），随后由丹麦Exiqon服务公司通过下一代测序技术完成microRNA表达谱的检测分析。