單細(xì)胞western blot解析乳腺癌發(fā)生過(guò)程的細(xì)胞異質(zhì)性

乳腺癌起源于乳腺上皮細(xì)胞，原因是乳腺上皮細(xì)胞發(fā)生基因改變，導(dǎo)致隨后組織穩(wěn)態(tài)喪失。科學(xué)家們經(jīng)過(guò)大量研究將乳腺上皮細(xì)胞分為幾個(gè)不同的亞群，但依然無(wú)法完全了解上皮細(xì)胞異質(zhì)性和分化譜。
 

加州大學(xué)歐文分校（University of California, Irvine）的科學(xué)家，在最新一期Nature Communication上發(fā)表文章，采用單細(xì)胞mRNA測(cè)序(ScRNAseq)分析來(lái)自7個(gè)不同乳腺增生病人的25,790個(gè)人乳腺上皮細(xì)胞的轉(zhuǎn)錄組，使用無(wú)偏聚分析揭示了三種不同的上皮細(xì)胞群的存在，一種是基底細(xì)胞，另一種是導(dǎo)管腔上皮細(xì)胞，分為激素分泌型L1和激素反應(yīng)L2型細(xì)胞。
文章作者從單細(xì)胞測(cè)序數(shù)據(jù)發(fā)現(xiàn)，KRT8在L2型細(xì)胞中比L1性表達(dá)水平高。

 

為了闡述KRT8蛋白質(zhì)表達(dá)異質(zhì)性，作者選擇了ProteinSimple Milo單細(xì)胞western blot。Milo可將細(xì)胞分為三個(gè)亞群：陰性，低表達(dá)，高表達(dá)。并可以準(zhǔn)確計(jì)算各亞群細(xì)胞的百分比即數(shù)量。

L2 was also characterized by higher levels of KRT8 than L1
 To quantify protein expression in individual cells, we utilized a recently developed single-cell western blot application (ProteinSimple, Milo), which performs electrophoretic separation of the protein content of about 2000 cells per chip and subsequently probed with fluorescently labeled antibodies.
Applying single-cell western blotting to luminal and basal cells isolated by FACS identified three cell states, namely KRT8-negative, -low, and -high , which illustrates the usefulness of single cell Western blotting as a quantitative validation tool downstream of scRNAseq analyses.

單細(xì)胞基因測(cè)序+Milo單細(xì)胞western blot組合，可以從基因水平和蛋白質(zhì)水平完整解析細(xì)胞異質(zhì)性，闡述腫瘤發(fā)生，干細(xì)胞發(fā)育等關(guān)鍵生命醫(yī)學(xué)問(wèn)題。