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Differentiation and maturation of human neuroblastoma cells into smooth muscle cells

人神经母细胞瘤细胞向平滑肌细胞的分化和成熟

基本信息

批准号：
12670763
负责人：
SUGIMOTO Tohru
金额：
$ 2.11万
依托单位：
Kyoto Prefectural University of Medicine
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2000
资助国家：
日本
起止时间：
2000 至 2001
项目状态：
已结题

项目摘要

Evidence that the neural crest generates peripheral neural cells, melanocytes and ectomesenchymal derivatives including smooth muscle cells, bone and cartilage has been demonstrated in avian embryos. Neuroblastoma is a tumor that is derived from the neural crest. Recent studies demonstrated that several human neuroblastoma cell lines exhibit at least three morphological types : neuroblastic (N)-type, substrate-adhesive (S)-type and intermediate (I)-type cells. However, the origin of the S-type cells has not been clearly identified. In this study, a total of six parent neuroblastoma cell lines (three N-types, two S-types and one mixed-type) and five cloned neuroblastoma cell lines (one N-type and four S-types) were used. The expressions of smooth muscle-specific proteins (desmin, α-smooth muscle act in, basic calponin and the smooth muscle myosin heavy-chain isoforms of SM1 and SM2) in neuroblastoma cell lines were analyzed. Three parent and four cloned neuroblastoma cell lines, compose … More d of S-type cells, were examined by indirect immunofluorescence, Western blot and/or by reverse transcriptase-polymerase chain reaction (RT-PCR). Desmin was found in two of the seven cell lines, and α-smooth muscle actin and basic calponin were detected in all of seven of the cell lines. In three parent cell lines and one cloned cell line composed of N-type cells, none of three smooth muscle-specific proteins were detected. In smooth muscle myosin heavy-chain isoforms, SM1 was detected in two parent cell lines composed of S-type cells (MP-N-MS and KP-N-YS) by immunofluorescence, Western blot and/or by RT-PCR, whereas the SM2 isoform was detected in one parent cell line (MP-N-MS) by RT-PCR. Neuroblastoma cell lines expressing basic calponin and isoforms of SM1 and SM2 have never been reported. MP-N-MS cells had the most mature smooth muscle phenotypes among the S-type neuroblastoma cell lines. These findings indicate that S-type cells have either the immature or mature smooth muscle cell phenotype, and neural crest cells very likely have the ability of to differentiate into smooth muscle cells in the human system. Less

在鸟类胚胎中已经证明神经嵴产生外周神经细胞、黑素细胞和外胚间充质衍生物，包括平滑肌细胞、骨和软骨。神经母细胞瘤是一种来源于神经嵴的肿瘤。近年来的研究表明，几种人神经母细胞瘤细胞系至少表现出三种形态学类型：神经母细胞（N）型、基质粘附（S）型和中间（I）型细胞。然而，S型细胞的起源尚未明确确定。在这项研究中，共使用了六个亲本神经母细胞瘤细胞系（三个N型，两个S型和一个混合型）和五个克隆的神经母细胞瘤细胞系（一个N型和四个S型）。分析神经母细胞瘤细胞系中平滑肌特异性蛋白（结蛋白、α-平滑肌肌动蛋白、碱性钙调蛋白和平滑肌肌球蛋白重链亚型SM 1和SM 2）的表达。三个亲本和四个克隆的神经母细胞瘤细胞系， ...更多信息 d的S型细胞，通过间接免疫荧光，Western blot和/或通过逆转录聚合酶链反应（RT-PCR）检测。在7个细胞系中的2个中发现结蛋白，并且在所有7个细胞系中检测到α-平滑肌肌动蛋白和碱性钙调蛋白。在三个亲本细胞系和一个由N型细胞组成的克隆细胞系中，没有检测到三种平滑肌特异性蛋白。在平滑肌肌球蛋白重链亚型中，通过免疫荧光、Western印迹和/或RT-PCR在由S型细胞组成的两个亲本细胞系（MP-N-MS和KP-N-YS）中检测到SM 1，而通过RT-PCR在一个亲本细胞系（MP-N-MS）中检测到SM 2亚型。神经母细胞瘤细胞株表达碱性钙调蛋白和亚型的SM 1和SM 2从未报道。在S型神经母细胞瘤细胞系中，MP-N-MS细胞具有最成熟的平滑肌表型。这些发现表明，S型细胞具有未成熟或成熟的平滑肌细胞表型，并且神经嵴细胞很可能具有在人体系统中分化为平滑肌细胞的能力。少