Differentially expressed miRNAs in TGF-β1-knock down rhabdmyosarcoma cell lines vs controls. Homo sapiens

To investigate the role of TGF-β1-regulated miRNAs in the progression of RMS,we performed comprehensive miRMA microarray analysis on RNA derived from typical RMS cell lines and TGF-β1 knock-down cell lines. We identified a novel set of TGF-β1-related miRNAs. Overall design: Total RNA was isolated from TGF-β1-knock down rhabdmyosarcoma cell lines and controls. Three-condition experiment: shRNA-TGF-β1/RD cells vs. shRNA-Control/RD cells, shRNA-TGF-β1/SMS-CTR cells vs. shRNA-Control/ SMS-CTR cells, and shRNA-TGF-β1/RH28 cells vs. shRNA-Control/RH28 cells. Biological replicates: 1 RD cells stably transfected with shRNA-TGF-β1- pSUPER gfp-neo, 1SMS-CTR cells stably transfected with shRNA-TGF-β1- pSUPER gfp-neo, 1RH28 cells stably transfected with shRNA-TGF-β1- pSUPER gfp-neo, 1RD cells stably transfected with shRNA-Control- pSUPER gfp-neo, 1SMS-CTR cells stably transfected with shRNA-Control- pSUPER gfp-neo, and 1RH28 cells stably transfected with shRNA-Control- pSUPER gfp-neo, independently grown and harvested. One replicate per array.

为探究转化生长因子β1（TGF-β1）调控的微小RNA（miRNAs）在横纹肌肉瘤（RMS）进展中的作用，我们针对典型横纹肌肉瘤细胞系及转化生长因子β1敲低细胞系来源的RNA开展了全面的miRNA微阵列分析，并鉴定出一组全新的与转化生长因子β1相关的微小RNA。
总体实验设计：总RNA提取自转化生长因子β1敲低的横纹肌肉瘤细胞系及其对照细胞系。本实验包含三组对照：短发卡RNA（short hairpin RNA, shRNA）-TGF-β1转染的RD细胞与shRNA-对照（shRNA-Control）转染的RD细胞、shRNA-TGF-β1转染的SMS-CTR细胞与shRNA-Control转染的SMS-CTR细胞，以及shRNA-TGF-β1转染的RH28细胞与shRNA-Control转染的RH28细胞。
生物学重复设置如下：分别独立培养并收集1株稳定转染shRNA-TGF-β1-pSUPER gfp-neo的RD细胞、1株稳定转染shRNA-TGF-β1-pSUPER gfp-neo的SMS-CTR细胞、1株稳定转染shRNA-TGF-β1-pSUPER gfp-neo的RH28细胞、1株稳定转染shRNA-Control-pSUPER gfp-neo的RD细胞、1株稳定转染shRNA-Control-pSUPER gfp-neo的SMS-CTR细胞，以及1株稳定转染shRNA-Control-pSUPER gfp-neo的RH28细胞；每块芯片对应一个生物学重复。