Differentiation and maturation of human neuroblastoma cells into smooth muscle cells

有证据表明，神经嵴产生周围神经细胞、黑素细胞和包括平滑肌细胞、骨和软骨在内的外胚间充质衍生物。神经母细胞瘤是一种源自神经嵴的肿瘤。最近的研究表明，几种人类神经母细胞瘤细胞系至少表现出三种形态类型：神经母细胞(N)型、底物粘附(S)型和中间(I)型细胞。然而，s型细胞的起源尚未明确。本研究共使用6株亲本神经母细胞瘤细胞系（3株n型、2株s型和1株混合型）和5株克隆神经母细胞瘤细胞系（1株n型和4株s型）。分析了成神经细胞瘤细胞株中平滑肌特异性蛋白（desmin、α-平滑肌行为蛋白、碱性钙钙蛋白和平滑肌肌球蛋白重链亚型SM1和SM2）的表达。采用间接免疫荧光、Western blot和/或逆转录聚合酶链反应（RT-PCR）对3株亲本和4株克隆的神经母细胞瘤细胞株进行了检测。7个细胞系中有2个检测到Desmin， α-平滑肌肌动蛋白和碱性钙蛋白均在7个细胞系中检测到。在3个亲本细胞系和1个由n型细胞组成的克隆细胞系中，均未检测到3种平滑肌特异性蛋白。在平滑肌肌球蛋白重链亚型中，SM1通过免疫荧光、Western blot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，而SM2则通过RT-PCR在一个亲本细胞系（MP-N-MS）中检测到。神经母细胞瘤细胞系表达碱性钙钙蛋白和SM1和SM2亚型从未报道过。在s型神经母细胞瘤细胞系中，MP-N-MS细胞平滑肌表型最成熟。这些发现表明，s型细胞既有未成熟的平滑肌细胞表型，也有成熟的平滑肌细胞表型，神经嵴细胞在人体系统中极有可能具有向平滑肌细胞分化的能力。少

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