Najafi Mostafa, Alizadeh Safar Ali, Azad Mehdi, Naserpour Farivar Taghi, Rajaei Farzad, Hotam Sorouri Kourosh, Rahmani Babak, Gheibi Nematollah
Department of Molecular Medicine, School of Medicine, Qazvin University of Medical Sciences , Qazvin , Iran.
Department of Medical Microbiology, School of Medicine, Qazvin University of Medical Sciences , Qazvin , Iran.
Turk J Biol. 2017 Dec 18;41(6):849-856. doi: 10.3906/biy-1704-14. eCollection 2017.
Increased levels of calprotectin subunits S100A8 and S100A9 have been detected in human cancers. Melanoma is the most aggressive type of skin cancer, and its treatment is challenging because of its brain metastasis. OCLN encodes occluding, which plays a major role in the formation and regulation of tight junctions. The aim of this study was to evaluate the methylation status of the OCLN promoter and its expression in A-375 melanoma cells treated with or without various concentrations of S100A9 for 24, 48, and 72 h. Total RNA was extracted, and synthesized cDNA was assessed by performing real-time PCR. MSP-PCR was performed after treatment with bisultfie. Recombinant S100A9 inhibited the proliferation of the A-375 cell line and the expression of the OCLN gene was downregulated in a time- and concentration-dependent manner. Results of MSP-PCR showed that the OCLN gene promoter in a human melanoma cell line (A-375) was semimethylated.
在人类癌症中已检测到钙卫蛋白亚基S100A8和S100A9水平升高。黑色素瘤是最具侵袭性的皮肤癌类型，由于其脑转移，其治疗具有挑战性。OCLN编码闭合蛋白，其在紧密连接的形成和调节中起主要作用。本研究的目的是评估OCLN启动子的甲基化状态及其在不同浓度S100A9处理24、48和72小时的A-375黑色素瘤细胞中的表达。提取总RNA，并通过实时PCR评估合成的cDNA。用亚硫酸氢盐处理后进行甲基化特异性PCR（MSP-PCR）。重组S100A9抑制A-375细胞系的增殖，并且OCLN基因的表达以时间和浓度依赖性方式下调。MSP-PCR结果显示人黑色素瘤细胞系（A-375）中的OCLN基因启动子是半甲基化的。
