Expression data from colon fibroblasts treated with Sonic hedgehog homolog (SHH)

Canonical Hedgehog (Hh) signaling regulates the expression of genes that are critical to the patterning and development of a variety of organ systems. In adult, both ligand-dependent and ligand-independent Hh pathway activation are known to promote tumorigenesis. Recent studies have shown that in tumors promoted by Hh ligand, activation occurs within the stromal microenvironment (Yauch et al., 2009). In situ hybridization of the pathway target gene, Ptch1, shows that signaling is located at stromal perivascular fibroblast-like cells in xenograft tumor sections derived from Hh-expressing colorectal cancer cell lines. To study the downstream genes regulated by Hh signaling, we treated a primary human colon myofibroblast, CCD-18Co, with SHH (1 ug/ml) or no treatment (control) in serum-free medium supplemented with 0.1% BSA for 72 hrs and performed microarray analysis (Affymetrix U133P) on these samples. Three biological replicates of SHH stimulated and three replicates of unstimulated primary colon myofibroblast cells CCD-18Co were used in the experiment to analyze their gene expression.

经典刺猬（Hedgehog, Hh）信号通路可调控多种器官系统模式形成与发育过程中关键基因的表达。在成年个体中，配体依赖性与配体非依赖性的Hh通路激活均被证实可促进肿瘤发生。近期研究表明，在Hh配体驱动的肿瘤中，信号激活发生于间质微环境内（Yauch等，2009）。对通路靶基因Ptch1的原位杂交实验显示，在表达Hh的结直肠癌细胞系构建的异种移植瘤组织切片中，信号活性定位于间质血管周成纤维样细胞。为研究Hh信号通路调控的下游基因，我们将原代人结肠肌成纤维细胞CCD-18Co置于添加0.1%牛血清白蛋白（BSA）的无血清培养基中，分别以1μg/ml的音猬因子（Sonic Hedgehog, SHH）处理或不做处理（作为对照组），持续培养72小时，随后对所有样本开展微阵列分析（Affymetrix U133P芯片）。本实验共设置3组SHH刺激的原代结肠肌成纤维细胞CCD-18Co生物学重复，以及3组未刺激的生物学重复，用于分析其基因表达谱。