Gene expression profiles of tumor-associated macrophages (TAMs) overexpressing miR-511-3p. Mus musculus

Hematopoietic stem/progenitor cells (HS/PCs) were transduced with lentiviral vectors overexpressing OFP and either miR-511-3p or a control, mutated miRNA sequence (miR-511-3p-mut). The transduced HS/PCs were then transplanted in recipient C57BL/6 mice. Tumors (Lewis lung carcinomas, LLC) were injected s.c. 4 weeks after HS/PC transplant. Lentiviral vector-transduced (OFP+), tumor-associated macrophages (TAMs) were isolated 4 weeks after LLC injection by fluorescence-activated cell sorting. Gene expression profiles of TAMs overexpressing either miR-511-3p or miR-511-3p-mut were obtained from 3 independent biological samples/each. Gene expression profiles of miRNA-overexpressing TAMs were then compared with the gene expression profiles of wild-type TAMs isolated from LLCs grown in nontransplanted C57BL/6 mice; the latter TAMs were subfractioned into MRC1+CD11c(low) or CD11c+MRC1(low) subsets before RNA isolation and analysis. Comparison of gene expression profiles of TAMs revealed that miR-511-3p overexpression tunes down the expression of multiple genes that are classically upregulated in protumoral MRC1+CD11c(low) TAMs. Overall design: TAMs were isolated from Lewis lung carcinomas grown s.c. in C57BL/6 mice.

造血干/祖细胞（Hematopoietic stem/progenitor cells, HS/PCs）经过表达OFP与miR-511-3p或对照突变型miRNA序列（miR-511-3p-mut）的慢病毒载体（lentiviral vectors）转导。将转导后的HS/PCs移植至受体C57BL/6小鼠体内。造血干/祖细胞移植4周后，于小鼠皮下注射路易斯肺癌（Lewis lung carcinomas, LLC）细胞。路易斯肺癌接种4周后，通过荧光激活细胞分选（fluorescence-activated cell sorting）分离慢病毒载体转导的（OFP阳性）肿瘤相关巨噬细胞（tumor-associated macrophages, TAMs）。分别从3份独立生物学样本中获取过表达miR-511-3p或miR-511-3p-mut的TAMs的基因表达谱。将过表达miRNA的TAMs的基因表达谱，与从未接受造血干/祖细胞移植的C57BL/6小鼠体内生长的路易斯肺癌中分离的野生型TAMs的基因表达谱进行比较；后者在进行RNA分离与分析前，被分为MRC1+CD11c(低表达)或CD11c+MRC1(低表达)两个亚群。对TAMs基因表达谱的比较分析显示，miR-511-3p过表达可下调促肿瘤性MRC1+CD11c(低表达) TAMs中经典上调的多个基因的表达。整体实验设计：从C57BL/6小鼠皮下生长的路易斯肺癌中分离肿瘤相关巨噬细胞。