Ocampo-Candiani Jorge, Salinas-Santander Mauricio, Trevino Victor, Ortiz-López Rocio, Ocampo-Garza Jorge, Sanchez-Dominguez Celia Nohemi
Dermatology Service, Hospital Universitario Dr. José Eleuterio González, Facultad de Medicina, Universidad Autónoma de Nuevo León, Nuevo León, México.
Department of Investigation, Facultad de Medicina Unidad Saltillo, Universidad Autónoma de Coahuila, Saltillo, México.
An Bras Dermatol. 2018 Nov/Dec;93(6):843-851. doi: 10.1590/abd1806-4841.20187589.
Vitiligo is characterized by a lack of pigmentation in the skin. To date, there are no studies that analyze the changes in gene expression in the skin of vitiligo patients in response to narrow-band ultraviolet B (nb-UVB) phototherapy treatment.
Explore the usefulness of new generation RNA sequencing in the identification of gene expression changes in the skin of vitiligo patients treated with nb-UVB phototherapy.
Four skin biopsies (4mm in diameter) were collected from 45 Mexican vitiligo vulgaris patients, 2 specimens before and 2 after treatment with nb-UVB phototherapy, obtained from pigmented and non-pigmented tissue. RNA extracted from the biopsies was analyzed using the Illumina TruSeq Targeted RNA Expression protocol to study the expression of genes that participate in pathways of skin homeostasis. The 2 groups were compared using Student's t-test and the Mann-Whitney U-test.
The expression analysis identified differences in 12 genes included in this study after comparing the samples obtained before and after treatment: 5 genes involved in skin pigmentation, 2 genes involved in apoptosis, 2 genes involved in cell survival, 2 genes involved in oxidative stress responses and 1 gene involved in signal transduction mechanisms (p<0.05).
The small size of skin biopsies limits the amount of RNA obtained, the number of genes to be analyzed and the use of conventional techniques such as RT-qPCR.
We demonstrated usefulness of new generation RNA sequencing in the identification of gene expression changes, in addition to identifying new targets in the study of vitiligo.
白癜风的特征是皮肤色素脱失。迄今为止，尚无研究分析白癜风患者皮肤在窄谱中波紫外线（nb-UVB）光疗治疗后的基因表达变化。
探讨新一代RNA测序在鉴定接受nb-UVB光疗的白癜风患者皮肤基因表达变化中的作用。
从45名墨西哥寻常型白癜风患者身上采集4块皮肤活检样本（直径4毫米），其中2块在nb-UVB光疗治疗前采集，2块在治疗后采集，分别取自色素沉着和非色素沉着组织。使用Illumina TruSeq靶向RNA表达方案对活检样本中提取的RNA进行分析，以研究参与皮肤稳态途径的基因表达。两组样本采用学生t检验和曼-惠特尼U检验进行比较。
通过对治疗前后样本的比较，表达分析确定了本研究中12个基因存在差异：5个基因参与皮肤色素沉着，2个基因参与细胞凋亡，2个基因参与细胞存活，2个基因参与氧化应激反应，1个基因参与信号转导机制（p<0.05）。
皮肤活检样本量小限制了RNA的获取量、可分析的基因数量以及逆转录定量聚合酶链反应（RT-qPCR）等传统技术的应用。
我们证明新一代RNA测序在鉴定基因表达变化方面有用，此外还在白癜风研究中鉴定了新的靶点。
