Expression data from OL and OC treated B16F10 cells

Oleocanthal (OL) and Oleacein (OC) are enriched components in Olive (oil and leaves). However, their effects on skin have not been widely studied compared to Oleuropein (OP). Here, we perfomed global gene expression profiling to screen the potential effects of OL and OC on the skin. Results showed that OL and OC have effects on skin development and keratinocyte differentiation by upregulation of key markers. Furthermore, OL and OC were effectively downregulated several melanogenic genes and it suggests that OL and OC can have a potential effect for pigmentation. B16F10 were murine melanoma cells line, treated with 5 µM of OL and OC for 24 hours. Microarray gene expression profiling was conducted technical replicates. RNA was extracted using Isogen (Nippon Gene Co. Ltd., Toyama, Japan). The integrity of RNA was quantified using NanoDrop 2000 spectrophotometer (Thermo Scientific, Wilmington, DE, USA). RNA samples were prepared for gene expression profiling analysis with GeneChip® 3' Expression Arrays using 3' IVT PLUS Reagent Kit (Affymetrix Inc., Santa Clara, CA, USA). Two hundred and fifty ng of total RNA from each sample was used to generate amplified and biotinylated complementary RNA (cRNA) from poly (A) RNA in a total RNA sample according to the user manual. IVT Incubation time was 16 hour. GeneAtlas® Hybridization, Wash and Stain Kit was used for hybridizing 3' IVT Array Strips according to the user manual (P/N 08-0306). The mouse genome array strips were hybridized for 16 hours in a 45oC incubator, washed and stained and finally imaging was done with the GeneAtlas Fluidics and Imaging Station.

油酰花青苷（Oleocanthal, OL）与油酰苦苷（Oleacein, OC）是橄榄（油与叶片）中的富集活性成分。然而相较于橄榄苦苷（Oleuropein, OP），二者对皮肤的生物学效应尚未得到广泛研究。本研究通过全基因组基因表达谱分析（global gene expression profiling），筛选OL与OC对皮肤的潜在调控作用。研究结果显示，OL与OC可通过上调关键标志物，调控皮肤发育与角质形成细胞分化过程。此外，OL与OC可有效下调多个黑素生成相关基因，提示二者对色素沉着具有潜在干预价值。本研究采用小鼠黑色素瘤细胞系B16F10（B16F10），以5 µM浓度的OL与OC处理细胞24小时。实验设置技术重复（technical replicates）后开展了微阵列基因表达谱分析（microarray gene expression profiling）。总RNA采用Isogen试剂（日本Nippon Gene Co. Ltd., 富山市）提取；RNA的完整性通过NanoDrop 2000分光光度计（NanoDrop 2000 spectrophotometer，美国赛默飞世尔科技公司，特拉华州威尔明顿市）进行定量检测。RNA样品依照操作手册，使用GeneChip® 3'表达阵列（GeneChip® 3' Expression Arrays）搭配3' IVT PLUS试剂套装（3' IVT PLUS Reagent Kit，美国Affymetrix公司，加利福尼亚州圣克拉拉市）制备用于基因表达谱分析。每个样品取250 ng总RNA，依据操作手册从总RNA中的poly(A) RNA扩增并合成生物素标记的互补RNA（complementary RNA, cRNA）。体外转录（IVT）孵育时长为16小时。采用GeneAtlas® 杂交、洗涤与染色试剂盒（GeneAtlas® Hybridization, Wash and Stain Kit，产品编号P/N 08-0306），依照操作手册对3' IVT阵列条带进行杂交。小鼠基因组阵列条带于45℃恒温孵箱中杂交16小时，经洗涤、染色后，通过GeneAtlas流体与成像工作站（GeneAtlas Fluidics and Imaging Station）完成成像扫描。