CD44(+)/CD24(-) breast cancer cells exhibit phenotypic reversion in three-dimensional self-assembling peptide RADA16 nanofiber scaffold.
CD44(+)/CD24(-) breast cancer cells exhibit phenotypic reversion in three-dimensional self-assembling peptide RADA16 nanofiber scaffold.
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DOI：
10.2147/ijn.s66723
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发表时间：
2015
影响因子：
8
通讯作者：
Xing Z
中科院分区：
文献类型：
--
作者：
Mi K;Xing Z
自组装肽支架已被证明是一种允许的生物材料，用于组织修复，细胞增殖，分化等。最近，乳腺癌细胞的亚群（CD 44 +/CD 24 −）已被报道具有干/祖细胞特性。本研究的目的是研究与Matrigel®（BD Biosciences，Two Oak Park，贝德福德，MA，USA）和胶原蛋白I相比，该癌细胞亚群在自组装COCH 3-RADARADARARADARADA-CONH 2（RADA 16）肽支架中是否具有不同的表型。流式细胞术测定CD 44和CD 24表达。5-溴-2 ′-脱氧尿苷法和DNA含量测定法检测细胞增殖情况。免疫染色用于指示不同支架的三维（3D）培养物中的细胞形态和集落中β-连环蛋白的定位。Western blot检测细胞内信号蛋白的表达。采用体外迁移实验和裸鼠体内接种实验评价其侵袭和成瘤性。乳腺癌细胞系MDA-MB-435 S含有高百分比（>99%）的CD 44 +/CD 24 −细胞，与胶原蛋白I和Matrigel相比，其在3D RADA 16支架中表现出表型逆转。新形成的回复腺泡样菌落重新组装基膜并重组其细胞骨架。同时，培养并包埋在RADA 16肽支架中的细胞表现出生长停滞。此外，它们表现出不同的迁移潜力，这将它们的迁移能力与它们的细胞形态联系起来。与体外研究一致，体内肿瘤形成测定进一步支持由3D RADA 16培养物中的回复引起的功能变化。RADA 16可诱导MDA-MB-453 S细胞表面粘附分子-1表达上调，NF-κ B B抑制剂吡咯烷二硫代氨基甲酸酯可抑制RADA 16诱导的MDA-MB-453 S细胞表面粘附分子-1表达上调及细胞表型逆转。与Matrigel和胶原I相比，在3D RADA 16肽支架中培养富含CD 44 +/CD 24 −的乳腺癌细胞群导致了显著的表型逆转。
Self-assembling peptide nanofiber scaffolds have been shown to be a permissive biological material for tissue repair, cell proliferation, differentiation, etc. Recently, a subpopulation (CD44+/CD24−) of breast cancer cells has been reported to have stem/progenitor cell properties. The aim of this study was to investigate whether this subpopulation of cancer cells have different phenotypes in self-assembling COCH3-RADARADARADARADA-CONH2 (RADA16) peptide nanofiber scaffold compared with Matrigel® (BD Biosciences, Two Oak Park, Bedford, MA, USA) and collagen I. CD44 and CD24 expression was determined by flow cytometry. Cell proliferation was measured by 5-bromo-2′-deoxyuridine assay and DNA content measurement. Immunostaining was used to indicate the morphologies of cells in three-dimensional (3D) cultures of different scaffolds and the localization of β-catenin in the colonies. Western blot was used to determine the expression of signaling proteins. In vitro migration assay and inoculation into nude mice were used to evaluate invasion and tumorigenesis in vivo. The breast cancer cell line MDA-MB-435S contained a high percentage (>99%) of CD44+/CD24− cells, which exhibited phenotypic reversion in 3D RADA16 nanofiber scaffold compared with collagen I and Matrigel. The newly formed reverted acini-like colonies reassembled a basement membrane and reorganized their cytoskeletons. At the same time, cells cultured and embedded in RADA16 peptide scaffold exhibited growth arrest. Also, they exhibited different migration potential, which links their migration ability with their cellular morphology. Consistent with studies in vitro, the in vivo tumor formation assay further supported of the functional changes caused by the reversion in 3D RADA16 culture. Expression levels of intercellular surface adhesion molecule-1 were upregulated in cells cultured in RADA16 scaffolds, and the NF-kappa B inhibitor pyrrolidine dithiocarbamate could inhibit RADA16-induced upregulation of intercellular surface adhesion molecule-1 and the phenotype reversion of MDA-MB-453S cells. Culturing a CD44+/CD24−-enriched breast cancer cell population in 3D RADA16 peptide nanofiber scaffold led to a significant phenotypic reversion compared with Matrigel and collagen I.