Shinohara Mayuka, Sumino Yasuhiro, Sato Fuminori, Kiyono Tohru, Hashimoto Naohiro, Mimata Hiromitsu
Department of Oncological Science (Urology), Oita University Faculty of Medicine, Yufu, Oita, Japan.
Division of Carcinogenesis and Cancer Prevention, National Cancer Center Research Institute, Tokyo, Japan.
Int J Urol. 2017 Jun;24(6):461-467. doi: 10.1111/iju.13330. Epub 2017 Mar 22.
To examine the inhibitory effects of tumor necrosis factor-α on myogenic differentiation of human urethral rhabdosphincter cells.
A rhabdosphincter sample was obtained from a patient who underwent total cystectomy. To expand the lifespan of the primary cultured cells, rhabdosphincter myogenic cells were immortalized with mutated cyclin-dependent kinase 4, cyclin D1 and telomerase. The differential potential of the cells was investigated. The transfected human rhabdosphincter cells were induced for myogenic differentiation with recombinant human tumor necrosis factor-α and/or the tumor necrosis factor-α antagonist etanercept at different concentrations, and activation of signaling pathways was monitored.
Human rhabdosphincter cells were selectively cultured for at least 40 passages. Molecular analysis confirmed the expression of myosin heavy chain, which is a specific marker of differentiated muscle cells, significantly increased after differentiation induction. Although tumor necrosis factor-α treatment reduced the myosin heavy chain expression in a concentration-dependent manner, etanercept inhibited this suppression. Tumor necrosis factor-α suppressed phosphorylation of protein kinase B and p38, whereas etanercept pretreatment promoted phosphorylation and myosin heavy chain expression in a concentration-dependent manner.
Tumor necrosis factor-α inhibits differentiation of urethral rhabdosphincter cells in part through the p38 mitogen-activated protein kinase and phosphoinositide 3-kinase pathways. Inhibition of tumor necrosis factor-α might be a useful strategy to treat stress urinary incontinence.
研究肿瘤坏死因子-α对人尿道横纹括约肌细胞肌源性分化的抑制作用。
从一名接受全膀胱切除术的患者身上获取横纹括约肌样本。为延长原代培养细胞的寿命，用突变的细胞周期蛋白依赖性激酶4、细胞周期蛋白D1和端粒酶使横纹括约肌肌源性细胞永生化。研究细胞的分化潜能。用不同浓度的重组人肿瘤坏死因子-α和/或肿瘤坏死因子-α拮抗剂依那西普诱导转染后的人横纹括约肌细胞进行肌源性分化，并监测信号通路的激活情况。
人横纹括约肌细胞选择性培养至少40代。分子分析证实，分化诱导后，分化肌肉细胞的特异性标志物肌球蛋白重链的表达显著增加。虽然肿瘤坏死因子-α处理以浓度依赖性方式降低了肌球蛋白重链的表达，但依那西普抑制了这种抑制作用。肿瘤坏死因子-α抑制蛋白激酶B和p38的磷酸化，而依那西普预处理以浓度依赖性方式促进磷酸化和肌球蛋白重链的表达。
肿瘤坏死因子-α部分通过p38丝裂原活化蛋白激酶和磷脂酰肌醇3-激酶途径抑制尿道横纹括约肌细胞的分化。抑制肿瘤坏死因子-α可能是治疗压力性尿失禁的一种有效策略。
