Experimental conditions and protein markers for redifferentiation of human coronary artery smooth muscle cells.

IF 2.3 Q3 MEDICINE, RESEARCH & EXPERIMENTAL
Ryota Shinozaki, Ryoji Eguchi, Ichiro Wakabayashi
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Abstract

A phenotype switch from contractile type to proliferative type of arterial smooth muscle cells is known as dedifferentiation, but to the best of our knowledge, little is known about redifferentiation of coronary artery smooth muscle cells. The purpose of the present study was to determine in vitro culture conditions for inducing redifferentiation of coronary artery smooth muscle cells. In addition, the present study aimed to determine protein markers for detection of redifferentiated arterial smooth muscle cells. Human coronary artery smooth muscle cells (HCASMCs) were cultured in the presence or absence of growth factors, including epidermal growth factor, fibroblast growth factor-B and insulin. Protein expression and migration activity of HCASMCs were evaluated using western blotting and migration assay, respectively. In HCASMCs 5 days after 100% confluency, expression levels of α-smooth muscle actin (α-SMA), calponin, caldesmon and SM22α were significantly increased, while expression levels of proliferation cell nuclear antigen (PCNA) and S100A4 and migration activity were significantly decreased, compared with the corresponding levels just after reaching 100% confluency, indicating that redifferentiation occurred. Redifferentiation was also induced in a low-density culture of HCASMCs in the medium without growth factors. When the culture medium for confluent cells was replaced daily with fresh medium, the expression levels of α-SMA, caldesmon, SM22α, PCNA and S100A4 and migration activity were not significantly different but the calponin expression was significantly increased compared with the levels in dedifferentiated cells just after reaching 100% confluency. Thus, redifferentiation was induced in HCASMCs by deprivation of growth factors from culture medium. The results suggested that α-SMA, caldesmon and SM22α, but not calponin, are markers of redifferentiation of HCASMCs.

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人冠状动脉平滑肌细胞再分化的实验条件及蛋白标记物。
动脉平滑肌细胞从收缩型到增生型的表型转换被称为去分化,但据我们所知,对冠状动脉平滑肌细胞的再分化知之甚少。本研究的目的是确定诱导冠状动脉平滑肌细胞再分化的体外培养条件。此外,本研究旨在确定检测再分化动脉平滑肌细胞的蛋白质标记物。人冠状动脉平滑肌细胞(HCASMCs)在有或没有生长因子的情况下培养,包括表皮生长因子、成纤维细胞生长因子- b和胰岛素。采用western blotting和迁移法分别评价HCASMCs的蛋白表达和迁移活性。在100%融合后5 d的HCASMCs中,α-平滑肌肌动蛋白(α-SMA)、calponin、caldesmon和SM22α的表达水平显著升高,增殖细胞核抗原(PCNA)和S100A4的表达水平和迁移活性显著降低,表明发生了再分化。在无生长因子的培养基中低密度培养HCASMCs也能诱导再分化。当融合细胞的培养基每天更换为新鲜培养基时,α-SMA、caldesmon、SM22α、PCNA和S100A4的表达量和迁移活性无显著差异,但钙钙蛋白的表达量在达到100%融合后较去分化细胞显著增加。因此,从培养基中剥夺生长因子可诱导HCASMCs再分化。结果表明α-SMA、caldesmon和SM22α是HCASMCs再分化的标志物,而calponin不是。
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来源期刊
Biomedical reports
Biomedical reports MEDICINE, RESEARCH & EXPERIMENTAL-
CiteScore
4.10
自引率
0.00%
发文量
86
期刊介绍: Biomedical Reports is a monthly, peer-reviewed journal, dedicated to publishing research across all fields of biology and medicine, including pharmacology, pathology, gene therapy, genetics, microbiology, neurosciences, infectious diseases, molecular cardiology and molecular surgery. The journal provides a home for original research, case reports and review articles.
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