Expression data from cells overexpressing c19orf63 (HSS1) [U87]. Homo sapiens

U87 cell lines were stable transfected with C19ORF63 (Human hematopoietic peptide secreted-1 - HSS1). HSS1 is a truly novel protein defining a new class of secreted factors. U87 cell line overexpressing HSS1 greatly reduced their proliferation rate compared to mock-transfected cells. Microarray analysis was used to detail gene expression underlying the anti-proliferative and anti-tumorigenic effect of HSS1 in U87 cells. Overall design: Exponentially growing U87 cells at growth curve day 5 were harvested for total RNA extraction and hybridization on Affimetrix microarrays. Three groups of samples were evaluated in triplicates: U87 wild-type, U87 -pcDNA3.1 mock-transfected, U87-pcDNA-HSS1. Cells were stable transfected with pcDNA3.1 empty vector or hHSS1. hHSS1-expressing cells and control cells were at confluence 40-80% when harvested. Trypan blue analysis of the number of viable cells showed a significant anti-proliferative effect in U87 cells expressing hHSS1 as compared to the control cells.

本研究中，U87细胞系经C19ORF63（人分泌型造血肽1，简称HSS1）稳定转染。HSS1是一种全新的蛋白质，定义了一类全新的分泌因子类别。与空载体转染的对照组细胞相比，过表达HSS1的U87细胞系增殖速率显著降低。本研究采用微阵列分析，详细解析了HSS1在U87细胞中发挥抗增殖及抗肿瘤生成效应的潜在基因表达调控机制。
实验整体设计如下：收集生长曲线第5天处于指数生长期的U87细胞，提取总RNA后开展Affimetrix微阵列杂交。实验共设置三组样本，每组均设置三次生物学重复：野生型U87细胞、U87-pcDNA3.1空载体转染组、U87-pcDNA-HSS1转染组。细胞分别以pcDNA3.1空载体或hHSS1进行稳定转染。收集hHSS1表达细胞与对照细胞时，二者的细胞汇合度均处于40%~80%区间。台盼蓝活细胞计数分析结果显示，相较于对照组细胞，表达hHSS1的U87细胞展现出显著的抗增殖效应。