4种哺乳动物细胞LRRC8A的表达及其生理特性的研究  

Expression of LRRC8Ain four kind of mammalian cells and study on its physiological characteristics

作　　者：张逸平[1] 韩杰 赵宏宇[2] ZHANG Yiping;HAN Jie;ZHAO Hongyu(Radiotherapy Department,First Inpatient Ward,China-Japan Union Hospital of Jilin University,Changchun 130033,China;Department of Radiotherapy,China-Japan Union Hospital of Jilin University,Changchun 130033,China)

机构地区：[1]吉林大学中日联谊医院放射治疗科一病区,吉林长春130033 [2]吉林大学中日联谊医院放射治疗科,吉林长春130033

出　　处：《中国实验诊断学》2025年第4期460-464,共5页Chinese Journal of Laboratory Diagnosis

基　　金：吉林省卫生科研人才专项项目(2023SCZ81);吉林大学中日联谊医院临床研究项目(YJZ2024020)。

摘　　要：目的研究4种哺乳动物细胞(人结肠癌细胞、人前列腺癌细胞、大鼠甲状腺滤泡上皮细胞、人卵巢癌细胞)中容积调控性阴离子通道LRRC8A的表达及其生理特性。方法RT-PCR检测LRRC8A在mRNA水平的表达,切胶回收PCR扩增产物进行基因测序并与比对;LRRC8A在蛋白质水平的表达应用Western Blot检验;构建YFP-H148Q/I152L真核表达载体,稳定转染至FRT细胞,荧光淬灭动力学实验验证低渗状态下LRRC8A的阴离子转运特性。结果RT-PCR及Western Blot表明以上4种哺乳动物细胞在mRNA和蛋白质水平均表达LRRC8A,基因测序结果经比对后显示被测序列与标准参考序列完全匹配。荧光淬灭动力学实验证实低渗状态下LRRC8A可转运阴离子,具有转运阴离子生理特性。结论4种哺乳动物细胞(人结肠癌细胞、人前列腺癌细胞、大鼠甲状腺滤泡上皮细胞、人卵巢癌细胞)表达LRRC8A,LRRC8A具有经典的容积调控性阴离子通道的特性。Objective To research the expression of LRRC8Ain four kind of mammalian cells(SW620/HCT116、PC3、FRT、SKOV3)and its physiological characteristics.Methods RT-PCR texted the expression of LRRC8Aat mRNA level,Western Blot was used to detect the expression at protein level,and PCR amplification products were recovered from gel cutting for gene sequencing and blast;YFP-H148Q/I152Lwas constructed and transfected stably into FRT cells,and the anion transport characteristics of LRRC8Ain low osmotic state were verified by fluorescence quenching kinetics experiment.Results RT-PCR and Western Blot showed that LRRC8Awas expressed in four kind of mammalian cells at mRNA and protein levels.Gene sequencing showed that the tested sequence matched the standard reference sequence.LRRC8Acould transport anions in low permeability state,and it has the characteristics of anion transport.Conclusion LRRC8Awas expressed in four kind of mammalian cells(SW620/HCT116、PC3、FRT、SKOV3)and had the classical characteristics of VRAC.

关 键 词：LRRC8A 容积调控性阴离子通道 哺乳动物细胞 FRT细胞 

分 类 号：R730.2[医药卫生—肿瘤]