Cancer Research UK Beatson Institute, Garscube Estate, Switchback Road, Glasgow G61 1BD, UK.
Institute of Cancer Sciences, University of Glasgow, Glasgow G12 8QQ, UK.
Sci Data. 2017 Nov 14;4:170172. doi: 10.1038/sdata.2017.172.
The metastatic spread of cancer cells is a step-wise process that starts with dissociation from primary tumours and local invasion of adjacent tissues. The ability to invade local tissues is the product of several processes, including degradation of extracellular matrices (ECM) and movement of tumour cells through physically-restricting gaps. To identify properties contributing to tumour cells squeezing through narrow gaps, invasive MDA-MB-231 human breast cancer and MDA-MB-435 human melanoma cells were subjected to three successive rounds of selection using cell culture inserts with highly constraining 3 μm pores. For comparison purposes, flow cytometry was also employed to enrich for small diameter MDA-MB-231 cells. RNA-Sequencing (RNA-seq) using the Illumina NextSeq 500 platform was undertaken to characterize how gene expression differed between parental, invasive pore selected or small diameter cells. Gene expression results obtained by RNA-seq were validated by comparing with RT-qPCR. Transcriptomic data generated could be used to determine how alterations that enable cell passage through narrow spaces contribute to local invasion and metastasis.
癌细胞的转移扩散是一个逐步的过程，始于与原发性肿瘤的分离和对邻近组织的局部侵袭。侵袭性是局部组织侵袭能力的产物，包括细胞外基质（ECM）的降解和肿瘤细胞通过物理限制间隙的运动。为了鉴定促进肿瘤细胞通过狭窄间隙的特性，采用具有高度限制 3 μm 孔的细胞培养插入物对侵袭性 MDA-MB-231 人乳腺癌和 MDA-MB-435 人黑色素瘤细胞进行了连续 3 轮选择。为了比较目的，还采用流式细胞术富集小直径 MDA-MB-231 细胞。使用 Illumina NextSeq 500 平台进行 RNA 测序（RNA-seq），以描述亲本、侵袭性孔选择或小直径细胞之间基因表达的差异。通过比较 RT-qPCR 对 RNA-seq 获得的基因表达结果进行验证。生成的转录组数据可用于确定促进细胞通过狭窄空间的改变如何有助于局部侵袭和转移。
