雄激素性秃发患者毛乳头细胞基因表达谱分析  

Genome-wide expression profile analysis of 3D cultured dermal papilla cells from patients with androgenic alopeeia

作　　者：庄晓晟[1] 孙蔚凌[1] 郑优优[1] 许嘉家[1] 胡莉芳[1] 范卫新[1] 

机构地区：[1]南京医科大学第一附属医院皮肤科,210029

出　　处：《中华皮肤科杂志》2014年第3期176-180,共5页Chinese Journal of Dermatology

基　　金：国家自然科学基金（81371754）;江苏省高校自然科学研究项目（11KJB320005）

摘　　要：目的探讨3D培养下，雄激素性秃发患者毛乳头细胞基因表达谱的差异。方法分离雄激素性秃发患者毛乳头细胞进行3D培养，实验组经双氢睾酮处理，提取总RNA进行扩增，Cy3标记，用NimbleGen人类全基因组表达谱芯片杂交，筛选差异表达基因进行GO分析及pathway分析，实时PCR对结果进行验证。结果全基因组表达谱分析显示，有622个基因有差异性表达，上调基因数359个，下调基因数为263个。其中GO分析显示，抑制细胞增殖、促进凋亡的基因出现上调，如CHEKl及Tobl等。促进细胞增殖、参与表皮生长发育的分子表达下调，如BAMBI、EFNA3、Dlx3及UCGC等。实时PCR验证结果与芯片结果的差异趋势一致。Pathway分析显示，调节细胞周期信号转导通路的分子富集在首位。结论雄激素性秃发发病受多种信号分子及信号转导通路调节，可能集中在调节细胞周期、细胞增殖及凋亡等方面。Objective To screen genes differentially expressed between dermal papilla cells from occipital and vertex scalp of patients with androgenic alopecia （AGA） through a 3D culture system. Methods Dermal papilla ceils isolated from the occipital scalp tissue of patients with AGA were cultured in a 2D system for several days. Then, the third-passage dermal papilla cells were subjected to a 3D culture with the presence of dihydrotestosterone （DHT） for 72 hours （experimental group）. The dermal papilla cells isolated from the vertex scalp tissue of patients with AGA, which were cultured in a 3D system with dimethyl sulfoxide, but not DHT, served as the control group. Subsequently, total RNA was extracted from the cells and reversely transcribed into cDNA followed by labeling with Cy3 and hybridization to a NimbleGen microarray. Gene ontology （GO） and pathway analysis was carried out to screen differentially expressed genes between the experimental and control group. Real time PCR was conducted to validate the results of microarray analysis. Results As the genome-wide expression profile analysis showed, there were 622 genes differentially expressed between the experimental group and control group, of which, 359 were up-regulated and 263 were down-regulated in the experimental group compared with the control group. The above results were confirmed by real time PCR. GO analysis revealed that the up-regulated genes, such as the CHEK1 and Tobl genes, were mainly involved in the inhibition of cell proliferation and promotion of cell apoptosis, while the down-regulated genes, such as the BAMBI, EFNA3, Dlx3 and UCGC genes, were associated with the acceleration of cell proliferation as well as the growth and development of epidermis. Pathway analysis showed that cell circle-controlling molecules were the most abundant molecules. Conclusions Numerous signalling molecules and pathways are involved in the development of AGA, which are mainly responsible for the modulation of cell circle, proliferation and apoptosis.

关 键 词：秃发 毛乳头细胞 细胞培养技术 寡核苷酸序列分析 

分 类 号：R758.71[医药卫生—皮肤病学与性病学]