Role of sEH R287Q in LDLR expression, LDL binding to LDLR and LDL internalization in BEL-7402 cells
Role of sEH R287Q in LDLR expression, LDL binding to LDLR and LDL internalization in BEL-7402 cells
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sEH R287Q 在 BEL-7402 细胞中 LDLR 表达、LDL 与 LDLR 结合以及 LDL 内化中的作用
DOI：
10.1016/j.gene.2018.04.038
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发表时间：
2018
期刊：
影响因子：
3.5
通讯作者：
Wang Luya
中科院分区：
文献类型：
--
作者：
Tang Ling;Wang Guoliang;Jiang Long;Chen Panpan;Wang Wei;Chen Juan;Wang Luya
目的家族性高胆固醇血症（FH）是一种常染色体显性遗传的胆固醇代谢紊乱。三种公认的基因（LDLR、APOB 和 PCSK9）仅存在于 20-30% 可能患有 FH 病例的患者中。需要探索其他导致 FH 的基因。本研究在 FH 核心家族中发现了一个孤立的基因变化 sEH R287Q。在本研究中，我们旨在研究R287Q对sEH表达和LDLR表达、LDL与LDLR结合以及LDL内化的作用。材料和方法使用基因捕获芯片对核心FH家族的167个脂质相关基因进行测序。通过载体依赖性蛋白表达，对培养的 BEL-7402 细胞中重组 sEH R287Q 的表达水平（蛋白质印迹）、水解酶活性（荧光化学）和细胞内定位（免疫荧光和共聚焦激光扫描显微镜）进行了研究。并通过流式细胞术检测野生型和sEH的R287Q对LDLR表达、LDL与LDLR结合以及LDL内化的影响。结果sEH R287Q是FH核心家族167个脂质相关基因中唯一的基因变化。与野生型sEH相比，重组sEH R287Q在培养细胞中的表达水平和水解酶活性均显着下降。 sEH R287Q 还降低 LDL 与 LDLR 的结合和 LDL 内化，并且对细胞表面 LDLR 蛋白水平没有影响。结论我们的结果表明 sEH R287Q 可能在 FH 血液 LDL 升高中发挥作用。 sEH R287Q 对 FH 的确切作用值得进一步研究。
ObjectivesFamilial hypercholesterolemia (FH) is an autosomal dominant disorder of cholesterol metabolism. Three recognized genes (LDLR, APOB and PCSK9) present in only 20–30% of patients with possible FH cases. Additional FH-causing genes need to be explored. The present study found an isolated gene change, sEH R287Q, in a core family of FH. In this study, we aimed to investigate the roles of R287Q on sEH expression and on LDLR expression, LDL binding to LDLR and LDL internalization.Materials and methods167 lipid-related genes of a core FH family were sequenced using a gene-capture chip. Through carrier dependent protein expression, the expression level (western blot), hydrolase activity (fluorescent chemistry) and intracellular localization (immunofluorescence and Confocal Laser Scanning Microscope) of recombinant sEH R287Q in cultured BEL-7402 cells were conducted. The effect of wild type and R287Q of sEH on LDLR expression, LDL binding to LDLR and LDL internalization were also conducted through Flow Cytometry.ResultssEH R287Q was the only gene changes among 167 lipid-related genes in the FH core family. Both expression level and hydrolase activity of recombinant sEH R287Q in cultured cells were significantly declined compared with that of the wild type sEH. sEH R287Q also decreased the binding of LDL to LDLR and LDL internalization and had no effect on cell-surface LDLR protein level.ConclusionOur results suggest that sEH R287Q may have a role in the elevation of blood LDL in FH. The exactly role of sEH R287Q on FH deserves further study.