miR-146a-5p在先天性巨中型黑色素细胞痣及正常皮肤中的差异表达

Differential expression of miR-146a-5p in giant congenital melanocytic nevi, medium-sized congenital melanocytic nevi and normal skin

目的 探讨先天性巨型黑色素细胞痣、先天性中型黑色素细胞痣和正常皮肤在miRNA表达水平的差异.方法 分别收集先天性巨型黑色素细胞痣、先天性中型黑色素细胞痣和正常皮肤样本各10例.第一步：每组选取3例样本进行AgilentmiRNA芯片检测,筛选不同组别间的差异miRNA.选取与MAPK、Wnt、NF-kB等信号通路有关的5个差异miRNA进行下一步验证,分别是：miR-146a-5p、miR-140-5p、miR-106b-5p、miR-17-5p、miR-483-5p.第二步：每组选取10例样本,通过实时荧光定量PCR（Taqman探针法）对上述差异miRNA表达进行定量验证并进行统计学分析.结果 通过miRNA芯片检测,共发现了先天性黑色素细胞痣与正常皮肤之间的23个差异miRNA.实时荧光定量PCR验证发现miR-146a-5p含量在先天性巨型黑色素细胞痣与中型黑色素细胞痣比较P=0.003;先天性巨型黑色素细胞痣与正常皮肤比较P=0.000;先天性中型黑色素细胞痣与正常皮肤对比P=0.013,三者之间差异均具有统计学意义.其他4个miRNA在3组间差异无统计学意义.结论 先天性黑色素细胞痣中miR-146a-5p的表达量增高,先天性巨型黑色素细胞痣增高尤为明显,这可能与不同组织中黑色素细胞的增殖和衰老状态有关.

Objective To investigate the differences in miRNA expression levels between giant congenital melanocytic nevi,medium-sized congenital melanocytic nevi and normal skin.Methods The experiment was divided into three groups：giant congenital melanocytic nevi group,medium-sized congenital melanocytic nevi group and normal skin group,with ten samples in each group.Firstly,3 samples of each group were detected by Agilent miRN Amicroarray to screen the different miRNAs between different groups.5 differential miRNAs related to MAPK,Wnt,NF-kB signaling pathways were selected for further verification：miR-146a-5p,miR-140-5p,miR-106b-5p,miR-17-5p,and miR-483-5p.miRNA expression levels were measured using real-time quantitative PCR （Taqman） in ten clinical samples from each group..Experimental data were analyzed using SPSS 22.0.Results A total of 23 differential miRNAs between congenital melanocytic nevi and normal skin were found by miRNA microarray detection.The results of real-time PCR showed that miR-146a-5p expression was significantly different between the three groups：giant congenital melanocytic nevi VS medium-sized congenital melanocytic nevi （P =0.003）;giant congenital melanocytic nevi VS normal skin （P =0.000）;medium-sized congenital melanocytic nevi VS normal skin （P =0.013）.There was no statistically significant difference in the expression of the other four miRNAs between the 3 groups.Conclusions The expression of miR-146a-5p is increased in congenital melanocytic nevi,especially in giant congenital melanocytic nevi,which may be related to the proliferation and senescence of melanocytes in different tissues.