Transcriptome analysis of Y79 retinoblastoma cells upon Lysyl oxidase propeptide (LOX-PP) overexpression. Transcriptome analysis of Y79 retinoblastoma cells upon Lysyl oxidase propeptide (LOX-PP) overexpression

Lysyl oxidase (LOX) is an copper dependent amine oxidase enzyme involved in cross linking of collagens and elastins. Lysyl oxidase propeptide (LOX-PP) is 18 kDa region cleaved during maturation of LOX and its anti-tumorigenic role were studied in various cancers. The effect of LOX-PP overexpression in retinoblastoma cancer cells (Y79) using transient transfection of LOX-PP gene accessed for global gene deregulations. The analysis resulted in RB cancer cell death through deregulation of retinoblastoma in cancer, cell cycle, apoptosis, focal adhesion-PI3K-AKT signaling and DNA repair mechanism pathway. Further validations were done using RT-PCR and western blot analysis. Our results provide evidence that inducing apoptosis through AKT-NFκB signaling. Overall design: We analyzed Y79 cell from 2 Empty vector and 2 LOX-PP overexpressed cells using the Affymetrix Human Transcriptome array 2.0 platform. Array data was processed by Affymetrix Transcriptome Analysis Console (TAC) 4.0 - Exon level Array Computational Tool. No techinical replicates were performed.

赖氨酰氧化酶（Lysyl oxidase, LOX）是一类铜依赖型胺氧化酶，参与胶原蛋白与弹性蛋白的交联修饰过程。赖氨酰氧化酶前肽（Lysyl oxidase propeptide, LOX-PP）是LOX成熟过程中被剪切释放的18 kDa肽段，其抗肿瘤活性已在多种癌症中得到研究。本研究通过将LOX-PP基因瞬时转染至视网膜母细胞瘤细胞（Y79）中，探究LOX-PP过表达对该细胞的影响，并分析全基因组表达失调情况。分析结果显示，通过失调癌症相关视网膜母细胞瘤（RB）通路、细胞周期、细胞凋亡、黏着斑-PI3K-AKT信号通路及DNA修复通路，可诱导视网膜母细胞瘤细胞死亡。后续通过反转录聚合酶链反应（RT-PCR）与蛋白质免疫印迹（Western blot）实验完成了验证。本研究结果证实，LOX-PP可通过AKT-NFκB信号通路诱导细胞凋亡。实验设计概述：我们采用Affymetrix人类转录组芯片2.0平台，对2株空载体转染的Y79细胞与2株LOX-PP过表达的Y79细胞进行了转录组分析。芯片数据通过Affymetrix转录组分析控制台（TAC）4.0——外显子水平阵列分析工具进行处理。本实验未设置技术重复。