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I型胶原蛋白作为CD133阳性胶质母细胞瘤细胞的潜在微环境成分。

Type 1 collagen as a potential niche component for CD133-positive glioblastoma cells.

作者信息

Motegi Hiroaki, Kamoshima Yuuta, Terasaka Shunsuke, Kobayashi Hiroyuki, Houkin Kiyohiro

机构信息

Department of Neurosurgery, Graduate School of Medicine, Hokkaido University, Sapporo, Japan.

出版信息

Neuropathology. 2014 Aug;34(4):378-85. doi: 10.1111/neup.12117. Epub 2014 Mar 27.

Abstract

Cancer stem cells are thought to be closely related to tumor progression and recurrence, making them attractive therapeutic targets. Stem cells of various tissues exist within niches maintaining their stemness. Glioblastoma stem cells (GSCs) are located at tumor capillaries and the perivascular niche, which are considered to have an important role in maintaining GSCs. There were some extracellular matrices (ECM) on the perivascular connective tissue, including type 1 collagen. We here evaluated whether type 1 collagen has a potential niche for GSCs. Imunohistochemical staining of type 1 collagen and CD133, one of the GSCs markers, on glioblastoma (GBM) tissues showed CD133-positive cells were located in immediate proximity to type 1 collagen around tumor vessels. We cultured human GBM cell lines, U87MG and GBM cells obtained from fresh surgical tissues, T472 and T555, with serum-containing medium (SCM) or serum-free medium with some growth factors (SFM) and in non-coated (Non-coat) or type 1 collagen-coated plates (Col). The RNA expression levels of CD133 and Nestin as stem cell markers in each condition were examined. The Col condition not only with SFM but SCM made GBM cells more enhanced in RNA expression of CD133, compared to Non-coat/SCM. Semi-quantitative measurement of CD133-positive cells by immunocytochemistry showed a statistically significant increase of CD133-positive cells in Col/SFM. In addition, T472 cell line cultured in the Col/SFM had capabilities of sphere formation and tumorigenesis. Type 1 collagen was found in the perivascular area and showed a possibility to maintain GSCs. These findings suggest that type 1 collagen could be one important niche component for CD133-positive GSCs and maintain GSCs in adherent culture.

摘要

癌症干细胞被认为与肿瘤进展和复发密切相关,这使得它们成为有吸引力的治疗靶点。各种组织的干细胞存在于维持其干性的微环境中。胶质母细胞瘤干细胞(GSCs)位于肿瘤毛细血管和血管周围微环境中,这被认为在维持GSCs方面具有重要作用。血管周围结缔组织上存在一些细胞外基质(ECM),包括1型胶原蛋白。我们在此评估1型胶原蛋白是否对GSCs具有潜在的微环境作用。对胶质母细胞瘤(GBM)组织进行1型胶原蛋白和GSCs标志物之一CD133的免疫组织化学染色显示,CD133阳性细胞位于肿瘤血管周围紧邻1型胶原蛋白的位置。我们用含血清培养基(SCM)或添加了一些生长因子的无血清培养基(SFM),并在未包被(Non - coat)或1型胶原蛋白包被的培养板(Col)中培养人GBM细胞系U87MG以及从新鲜手术组织获得的GBM细胞T472和T555。检测了每种条件下作为干细胞标志物的CD133和巢蛋白的RNA表达水平。与未包被/SCM相比,Col条件下无论是SFM还是SCM都使GBM细胞的CD133 RNA表达增强。通过免疫细胞化学对CD133阳性细胞进行半定量测量显示,Col/SFM中CD133阳性细胞有统计学意义的增加。此外,在Col/SFM中培养的T472细胞系具有形成球体和致瘤的能力。在血管周围区域发现了1型胶原蛋白,显示出维持GSCs的可能性。这些发现表明,1型胶原蛋白可能是CD133阳性GSCs的一个重要微环境成分,并在贴壁培养中维持GSCs。

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