Overall expression of genes in CRYM overexpressed Prostate cancer cell line LNCaP

RNA sequencing was employed to profile the entire mRNA transcripts. Total RNA from LNCaP transfected with control vector or CRYM bearing vector was isolated. RNA and samples were subjected to the sequencing on illumina's HiSeq 4000 platform. RNA-seq revealed the expression of 58,540 transcripts and of 34,878 (59.6 %) genes were expressed in LNCaP cells. 3226 (9.25 %) genes were altered by CRYM and 2.72 % were upregulated and 6.53 % were downregulated. Cells were in transiently transfected cells with CRYM vector vs control in duplicate. GFP expression was monitored and total RNA was isolated at 48 hours upon transfection.

本研究采用RNA测序（RNA sequencing）技术对全部mRNA转录本进行转录组分析。分别提取转染空载体或携带CRYM载体的LNCaP细胞中的总RNA，将上述样品置于Illumina的HiSeq 4000测序平台进行测序。RNA测序结果显示，共检测到58540个转录本的表达，其中34878个基因（占比59.6%）在LNCaP细胞中存在表达。3226个基因（占比9.25%）的表达受CRYM调控发生改变，其中上调基因占2.72%，下调基因占6.53%。本实验采用瞬时转染体系，设置CRYM载体转染组与空载体对照组，每组均设置两个生物学重复；于转染后48小时监测绿色荧光蛋白（GFP）的表达情况，并收集细胞以提取总RNA。