Department of Medical Genetics, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran.
Australian Regenerative Medicine Institute, Monash University, Clayton, VIC 3800, Australia.
Arch Iran Med. 2020 Oct 1;23(10):688-696. doi: 10.34172/aim.2020.87.
Ankylosing spondylitis (AS; OMIM:106300) is a common complex inflammatory disease; in a previous study, we introduced a novel mutation in the gene (OMIM: 600514) which was associated with AS. This study is designed to investigate the potential effect of S2486G mutation on reelin secretion; additionally, we objected to evaluate the phospholipase A2 () gene (OMIM: 601690) expression and platelet-activating factor-acetylhydrolase (PAF-AH) concentration as the downstream gene and the encoded protein.
The impact of the S2486G on reelin protein secretion was investigated in CHO-K1 and HEK-293T cells by constructing wild-type and mutant plasmids. Besides, the possible effect of the mutation on expression and concentration of and PAF-AH in THP1 cells was assessed by quantitative real-time PCR (qRT-PCR) and enzyme-linked immunosorbent assay (ELISA), respectively. The study was performed at Tarbiat Modares University, Tehran, Iran, from 2016 to 2018.
Our results showed that S2486G not only causes a significant reduction in reelin secretion in both HEK-293T and CHO-K1 cells, but also it leads to a significant reduction in gene expression ( value < 0.001) and protein level of PAF-AH in THP-1 cells ( value < 0.003).
The S2486G mutation in can alter inflammatory and, to some extent, osteogenesis pathways mediated by reduced secretion of reelin and also reduced expression of the gene.
强直性脊柱炎（AS；OMIM：106300）是一种常见的复杂炎症性疾病；在之前的研究中，我们介绍了基因（OMIM：600514）中的一个新突变与 AS 相关。本研究旨在探讨 S2486G 突变对 reelin 分泌的潜在影响；此外，我们旨在评估磷脂酶 A2（）基因（OMIM：601690）表达和血小板激活因子乙酰水解酶（PAF-AH）浓度作为下游基因和编码蛋白。
通过构建野生型和突变质粒，在 CHO-K1 和 HEK-293T 细胞中研究 S2486G 对 reelin 蛋白分泌的影响。此外，通过定量实时 PCR（qRT-PCR）和酶联免疫吸附试验（ELISA）分别评估突变对 THP1 细胞中表达和浓度的可能影响。该研究于 2016 年至 2018 年在伊朗德黑兰的塔比阿特莫达勒斯大学进行。
我们的结果表明，S2486G 不仅导致 HEK-293T 和 CHO-K1 细胞中 reelin 分泌显著减少，而且还导致 THP-1 细胞中基因表达（值<0.001）和 PAF-AH 蛋白水平显著降低（值<0.003）。
基因中的 S2486G 突变可以改变炎症和在某种程度上成骨途径，这是由于 reelin 分泌减少和基因表达减少引起的。
