circRNA CPSF6核内转移促进同型半胱氨酸诱导的滋养细胞凋亡  

The nuclear translocation of circRNA CPSF6 promotes apoptosis of trophoblast cells induced by homocysteine

作　　者：高丽娜 吴琪瑞 殷荷 刘林英 吴玉珠 张玉月 王艳华 吴凯[3] 张慧萍[2,4,5] GAO Lina;WU Qirui;YIN He;LIU Linying;WU Yuzhu;ZHANG Yuyue;WANG Yanhua;WU Kai;ZHANG Huiping(Department of Obstetrics and Gynecology,Clinical Medical College of Ningxia Medical University;Department of Obstetrics and Gynecology,General Hospital of Ningxia Medical University;Department of Histology and Embryology,Basic Medical College of Ningxia Medical University;Key Laboratory of Metabolic Cardiovascular Diseases Research,National Health Commission;Ningxia Key Laboratory of Vascular Injury and Repair Research,Yinchuan 750004,China)

机构地区：[1]宁夏医科大学临床医学院妇产科 [2]宁夏医科大学总医院妇产科 [3]宁夏医科大学基础医学院组胚学教研室 [4]国家卫生健康委代谢性心血管疾病研究重点实验室 [5]宁夏血管损伤与修复研究重点实验室,宁夏银川750004

出　　处：《细胞与分子免疫学杂志》2022年第2期146-152,共7页Chinese Journal of Cellular and Molecular Immunology

基　　金：国家自然科学基金(81760270);宁夏回族自治区重点研发计划(2019BFG02004,2018BEG03026);宁夏自然科学基金(2020AAC02038,2020AAC03380)

摘　　要：目的探讨环状RNA剪切及多聚腺苷酸化特异因子6(circRNA CPSF6)在同型半胱氨酸(Hcy)致滋养细胞凋亡中的作用及机制。方法体外培养HTR-8/SVneo人绒毛膜滋养层细胞,分为对照组(0 mmol/L Hcy处理)与1 mmol/L Hcy处理组。采用免疫荧光细胞化学染色检测滋养细胞胱天蛋白酶3(caspase-3)的表达,Western blot法检测caspase-3蛋白水平;实时定量PCR检测circRNA CPSF6的mRNA表达水平;同时,运用小干涉RNA(siRNA)技术敲低滋养细胞circRNA CPSF6表达,Western blot法检测细胞caspase-3、caspase-9、B细胞淋巴瘤分子2(Bcl2)、Bcl2相关X蛋白(BAX)表达;实时定量PCR检测circRNA CPSF6在Hcy处理前后滋养细胞胞质/胞核中的表达水平。结果与对照组相比,Hcy处理组caspase-3表达明显增加,circRNA CPSF6 mRNA表达上调;敲低circRNA CPSF6后,caspase-3表达降低,线粒体凋亡途径被抑制;正常培养的滋养细胞中circRNA CPSF6在胞质大量表达,而给予Hcy处理后,circRNA CPSF6主要在胞核表达。结论通过circRNA CPSF6核内转移激活线粒体凋亡途径促进Hcy诱导的滋养细胞凋亡。Objective To investigate the role of circular RNA cleavage and polyadenylation specificity factor 6(circRNA CPSF6) in the apoptosis of trophoblast cells induced by homocysteine(Hcy) and its mechanism. Methods HTR-8/SVneo human chorionic trophoblast cells were cultured in vitro and divided into control group(0 mmol/L Hcy treatment) and 1 mmol/L Hcy treatment group. Immunofluorescence cytochemical staining was used to detect the expression of caspase-3 in trophoblasts, and Western blot analysis was used to detect the caspase-3 protein level. The mRNA expression level of circRNA CPSF6 was detected by real-time quantitative PCR. Small interfering RNA(siRNA) was used to knock down the expression of circRNA CPSF6 in trophoblast cells. The expressions of caspase-3, caspase-9, Bcl2, and BAX were detected by Western blot analysis. Real-time quantitative PCR was used to detect the expression level of circRNA CPSF6 in the cytoplasm/nucleus of trophoblast cells before and after Hcy treatment. Results Compared with those in the control group, the expressions of caspase-3 and circRNA CPSF6 mRNA in the Hcy treatment group significantly increased. After knocking down circRNA CPSF6, the expression of caspase-3 decreased, and the mitochondrial apoptosis pathway was inhibited. In normal cultured trophoblast cells, circRNA CPSF6 was expressed in large amounts in the cytoplasm, and after Hcy treatment, circRNA CPSF6 was mainly expressed in the nucleus. Conclusion The mitochondrial apoptotic pathway is activated by circRNA CPSF6 nuclear translocation to promote trophoblast apoptosis induced by Hcy.

关 键 词：环状RNA剪切及多聚腺苷酸化特异因子6(circRNA CPSF6) 滋养细胞 同型半胱氨酸(Hcy) 细胞凋亡 

分 类 号：Q255[生物学—细胞生物学] R714.244[医药卫生—妇产科学]