hsa_circ_0008039和miR-484在SK-N-SH细胞氧糖剥夺-复糖复氧损伤中的作用:与Fis1的关系  

Role of hsa_circ_0008039 and miR-484 in oxygen-glucose deprivation/reoxygenation injury in SK-N-SH cells: relationship with Fis1

作　　者：袁阳 于洋[1] 孙洪喜 张高峰 陈怀龙[1] 王明山[1] Yuan Yang;Yu Yang;Sun Hongxi;Zhang Gaofeng;Chen Huailong;Wang Mingshan(Department of Anesthesiology,The Affiliated Hospital of Qingdao University,Qingdao 266071,China;Department of Anesthesiology,Jiaozhou Hospital Affiliated to Dongfang Hospital of Tongji University,Qingdao 266300,China)

机构地区：[1]青岛大学附属青岛市市立医院麻醉科,青岛266071 [2]同济大学附属东方医院胶州医院麻醉科,青岛266300

出　　处：《中华麻醉学杂志》2022年第1期88-92,共5页Chinese Journal of Anesthesiology

基　　金：山东省自然科学基金(ZR2021MH365)。

摘　　要：目的评价hsa_circ_0008039和miR-484在人神经母细胞瘤(SK-N-SH)细胞氧糖剥夺-复糖复氧(OGD/R)损伤中的作用及其与线粒体分裂蛋白1(Fis1)的关系。方法体外培养SK-N-SH细胞至对数生长期,采用随机数字表法分为5组(n=25):对照组(C组)、OGD/R组、hsa_circ_0008039 siRNA组(S组)、hsa_circ_0008039过表达组(E组)和hsa_circ_0008039 siRNA+miR-484抑制剂组(S+I组)。C组细胞在正常条件下培养;S组、E组和S+I组分别转染has_circ_0008039 siRNA、has_circ_0008039过表达载体、has_circ_0008039 siRNA和miR-484抑制剂后,氧糖剥夺12 h复糖复氧24 h制备OGD/R损伤模型。于复糖复氧24 h时采用CCK-8法检测细胞活力和LDH漏出量,采用RT-qPCR法检测hsa_circ_0008039、miR-484和Fis1 mRNA的表达,Western blot法检测Fis1表达;双荧光素酶报告验证hsa_circ_0008039与miR-484的靶向关系。结果与C组比较,其余4组细胞活力降低,LDH漏出量增加,OGD/R组和E组hsa_circ_0008039和Fis1表达上调,miR-484表达下调,S组hsa_circ_0008039和Fis1表达下调,miR-484表达上调,S+I组hsa_circ_0008039和miR-484表达下调,Fis1表达上调(P<0.05);与OGD/R组比较,E组和S+I组细胞活力降低,LDH漏出量增加,S组细胞活力升高,LDH漏出量减少,E组hsa_circ_0008039和Fis1表达上调,miR-484表达下调,S组hsa_circ_0008039和Fis1表达下调,miR-484表达上调,S+I组hsa_circ_0008039和miR-484表达下调,Fis1表达上调(P<0.05);与S组比较,S+I组细胞活力降低,LDH漏出量增加,miR-484表达下调,Fis1表达上调(P<0.01)。双荧光素酶报告实验表明:has_circ_0008039靶向结合miR-484。结论hsa_circ_0008039靶向结合miR-484参与了SK-N-SH细胞氧糖剥夺-复糖复氧损伤的过程,与上调Fis1表达有关。Objective To evaluate the role of has_circ_0008039 and miR-484 in oxygen-glucose deprivation/reoxygenation(OGD/R)injury in SK-N-SH cells and the relationship with Fis1.Methods SK-N-SH cells were cultured in vitro to logarithmic growth stage and divided into 5 groups(n=25 each)according to the random number table method:control group(group C),OGD/R group,has_circ_0008039 siRNA group(group S),hsa_circ_0008039 over-expression group(group E)and has_circ_0008039 siRNA plus miR-484 inhibitor group(group S+I).Cells were cultured in normal condition in group C.In S,E and S+I groups,after the cells were transfected with hsa_circ_0008039 siRNA,has_circ_0008039 over-expression vector,hsa_circ_0008039 siRNA and miR-484 inhibitor,the cells were subjected to oxygen-glucose deprivation for 12 h followed by 24 h restoration of O2-glucose supply to develop the OGD/R model.At 24 h of restoration of O2-glucose supply,the cell viability and amount of lactic dehydrogenase(LDH)released were measured using CCK-8 assay,the expression of hsa_circ_0008039,miR-484 and Fis1 mRNA was detected using real-time polymerase chain reaction,and the expression of Fis1 was detected by Western blot.A dual-fluorescein experimental report was used to verify the targeting relationship between hsa_circ_0008039 and miR-484.Results Compared with group C,the cell viability was significantly decreased,and the amount of LDH released was increased in the other 4 groups,the expression of hsa_circ_0008039 and Fis1 was significantly up-regulated,and the expression of miR-484 was down-regulated in OGD/R and E groups,the expression of hsa_circ_0008039 and Fis1 was significantly down-regulated,and miR-484 was up-regulated in group S,and the expression of hsa_circ_0008039 and miR-484 was significantly down-regulated,and the expression of Fis1 was up-regulated in group S+I(P<0.05).Compared with group OGD/R,the cell viability was significantly decreased,and the amount of LDH released was increased in E and S+I groups,the cell viability was significantly increased,and th

关 键 词：微RNAS 神经母细胞瘤 肿瘤细胞 培养的 低氧 线粒体蛋白质类 

分 类 号：R614[医药卫生—麻醉学]