| Co-Investigator(Kenkyū-buntansha) |
TAKAHASHI Atsushi Sapporo Medical University School of Medicine, Senior Instructor, 医学部, 助手 (20274946)
MASUMORI Naoya Sapporo Medical University School of Medicine, Assistant Professor, 医学部, 講師 (20295356)
ITOH Naoki Sapporo Medical University School of Medicine, Associate Professor, 医学部, 助教授 (60193504)
KUNISHIMA Yasuharu Sapporo Medical University School of Medicine, Senior Instructor, 医学部, 助手 (00315508)
SASAMURA Hiroto Sapporo Medical University School of Medicine, Senior Instructor, 医学部, 助手 (50311890)
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| Research Abstract |
In order to study the effects of TGF-βs on human prostatic stromal cells, we purified human prostatic myofibroblast using specimens that was taken by operations and established culture system of these cells. These cells were immunohistochemically recognized as myofibroblast according to the findings of positive staining of a-smooth muscle actin and negative staining of both myosin and desmin. The expressions of TCF-β receptor I, II mRNA were confirmed by RT-PCR in these cells. After the treatment of various concentrations of human TGF-β1, 2, 3, it was recognized that these cells expressed both myosin and desmin. These findings suggested that TGF-βs could differentiate myofibroblast to smooth muscle. On the other hand, TGF-β1, 2, 3 inhibited proliferation of myofibroblast by dose-dependent manner.
It has been well known that there are three subtypes of α1-receptor in human prostatic tissue; 1a, 1b and 1d, however the distribution of these subtypes in myofibroblast and smooth muscle cells
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has not been clarified. Using myofibroblast that was established by us and human prostatic smooth muscle that was commercially available, the expressions of 1a, 1b, 1d subtype mRNA were quantitatively analyzed by real time RT-PCR. The expression of 1a was recognized in smooth muscle only, and both 1b and 1d were determined in myofibroblast and smooth muscle cells. Based on these findings, it was speculated that the expression of ala receptor mRNA was induced during the differentiation from myofibroblast to smooth muscle.
To clarify the effect of TGF-β1 on the expression of α1 receptor in myofibroblast and smooth muscle, the expressios of α1a, 1b, 1d receptor mRNA were quantitatively analyzed by real time RT-PCR. Moreover, α1 receptor binding assay was performed using [^<125 >I] HEAT as a ligand. The expression of ala was detected in these cells cultured under the condition without TGF-β1. The expressions of 1b and 1d without TGF-β were significantly higher 0.13 and 0.09 times respectively than with TGF-β1. Bmax of [^^<125>I]HEAT (300pM) was 37.5 fmol/mg protein in the medium without TGF-β1 and 21.3fmol/mg protein in those with TGF-β1.
It was concluded that TGF-βs could have human prostatic myofibroblast differentiate to smooth muscle cell. On the other hand, the function of smooth muscle was undifferentiated by TGF-β1, because the expression of al receptor was suppressed by it. Based on these results, it was speculated that the regulatory mechanisms of morphological differentiation might be different from these of functional differentiation in human prostatic stroma. Less
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