Analysis of gene expression in co-culture of PBMCs with ovarian cancer cell lines by High-Throughput Sequencing. Analysis of gene expression in co-culture of PBMCs with ovarian cancer cell lines by High-Throughput Sequencing

(Objectives) The goal of this study is to investigate paracrine effect of peripheral blood mononuclear cells (PBMCs) on ovarian cancer cell lines. (Methods) Transcription profiles of PBMC (control) and PBMC co-cultured with ovarian cancer cell lines (treatment) were generated by deep sequencing, in triplicate, using Illumina HiSeq X Ten. (Results and Conclusions) Transcriptome analysis of PBMCs has revealed the expression change when co-cultured with EOC cell lines with significantly in up-regulated genes; CDKN1B, GIMAP8 and SNN. The qRT-PCR validation have indicated that GIMAP8 was high expressed in EOC patients when compared with healthy female. We summarized that PBMCs were changed their expression as a result of paracrine signals from ovarian cancer cells. Overall design: Transcription profiles of PBMC (control) and PBMC co-cultured with cancer (treatment) were generated by deep sequencing, using Illumina HiSeq 4000.

【研究目标】本研究旨在探究外周血单个核细胞（peripheral blood mononuclear cells, PBMCs）对卵巢癌细胞系的旁分泌效应。【实验方法】以单独培养的外周血单个核细胞作为对照组，以与卵巢癌细胞系共培养的外周血单个核细胞作为处理组，采用Illumina HiSeq X Ten平台完成三次生物学重复的深度测序，获取两组的转录组特征谱。【结果与结论】对共培养的外周血单个核细胞进行转录组分析后发现，其与上皮性卵巢癌（Epithelial Ovarian Cancer, EOC）细胞系共培养时基因表达发生显著改变，其中CDKN1B、GIMAP8及SNN基因显著上调。经实时荧光定量PCR（quantitative real-time polymerase chain reaction, qRT-PCR）验证显示，相较于健康女性，GIMAP8基因在上皮性卵巢癌患者体内呈高表达状态。本研究总结得出：外周血单个核细胞的基因表达改变源于卵巢癌细胞分泌的旁分泌信号。【整体实验设计】以单独培养的外周血单个核细胞作为对照组，以与癌细胞共培养的外周血单个核细胞作为处理组，采用Illumina HiSeq 4000平台进行深度测序，获取两组的转录组特征谱。