| Budget Amount *help |
¥3,300,000 (Direct Cost: ¥3,300,000)
Fiscal Year 2001: ¥1,200,000 (Direct Cost: ¥1,200,000)
Fiscal Year 2000: ¥2,100,000 (Direct Cost: ¥2,100,000)
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| Research Abstract |
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
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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
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