Sharp Julie A, Lefèvre Christophe, Watt Ashalyn, Nicholas Kevin R
Institute for Frontier Materials, Deakin University, Geelong, VIC, 3216, Australia.
Department of Anatomy and Developmental Biology, Monash University, Clayton, VIC, 3800, Australia.
Funct Integr Genomics. 2016 May;16(3):297-321. doi: 10.1007/s10142-016-0485-0. Epub 2016 Feb 24.
The molecular processes underlying human milk production and the effects of mastitic infection are largely unknown because of limitations in obtaining tissue samples. Determination of gene expression in normal lactating women would be a significant step toward understanding why some women display poor lactation outcomes. Here, we demonstrate the utility of RNA obtained directly from human milk cells to detect mammary epithelial cell (MEC)-specific gene expression. Milk cell RNA was collected from five time points (24 h prepartum during the colostrum period, midlactation, two involutions, and during a bout of mastitis) in addition to an involution series comprising three time points. Gene expression profiles were determined by use of human Affymetrix arrays. Milk cells collected during milk production showed that the most highly expressed genes were involved in milk synthesis (e.g., CEL, OLAH, FOLR1, BTN1A1, and ARG2), while milk cells collected during involution showed a significant downregulation of milk synthesis genes and activation of involution associated genes (e.g., STAT3, NF-kB, IRF5, and IRF7). Milk cells collected during mastitic infection revealed regulation of a unique set of genes specific to this disease state, while maintaining regulation of milk synthesis genes. Use of conventional epithelial cell markers was used to determine the population of MECs within each sample. This paper is the first to describe the milk cell transcriptome across the human lactation cycle and during mastitic infection, providing valuable insight into gene expression of the human mammary gland.
由于获取组织样本存在局限性，人乳生成背后的分子过程以及乳腺炎感染的影响在很大程度上尚不明确。确定正常哺乳期女性的基因表达将是朝着理解为何有些女性泌乳结果不佳迈出的重要一步。在此，我们证明了直接从人乳细胞中获取的RNA在检测乳腺上皮细胞（MEC）特异性基因表达方面的实用性。除了一个包含三个时间点的 involution 系列外，还从五个时间点（初乳期产前24小时、泌乳中期、两次 involution、乳腺炎发作期间）收集了乳细胞RNA。通过使用人类Affymetrix阵列确定基因表达谱。在乳汁生成期间收集的乳细胞显示，表达最高的基因参与乳汁合成（例如，CEL、OLAH、FOLR1、BTN1A1和ARG2），而在 involution 期间收集的乳细胞显示乳汁合成基因显著下调以及 involution 相关基因（例如，STAT3、NF-kB、IRF5和IRF7）激活。在乳腺炎感染期间收集的乳细胞揭示了一组特定于这种疾病状态的独特基因的调控，同时维持乳汁合成基因的调控。使用传统上皮细胞标记物来确定每个样本中MEC的群体。本文首次描述了人类泌乳周期以及乳腺炎感染期间的乳细胞转录组，为人类乳腺的基因表达提供了有价值的见解。
