微小RNA-K1-5p调控G1/S期相关基因的表达影响内皮细胞周期  

Effect of miR-K1-5p on the cycle of endothelial cells by regulating G1/S phase related gene expressions

作　　者：张静[1] 彭靖淇[2] ZHANG Jing;PENG Jingqi(Department of Pathology,The Affiliated Traditional Chinese Medicine Hospital,Xinjiang Medical University,Urumqi,Xinjiang Uygur Autonomous Region 830000,China;Department of Surgery,The Affiliated Traditional Chinese Medicine Hospital,Xinjiang Medical University,Urumqi,Xinjiang Uygur Autonomous Region 830000,China)

机构地区：[1]新疆医科大学附属中医医院病理科,新疆维吾尔自治区乌鲁木齐830000 [2]新疆医科大学附属中医医院外科,新疆维吾尔自治区乌鲁木齐830000

出　　处：《安徽医药》2023年第1期108-112,共5页Anhui Medical and Pharmaceutical Journal

基　　金：新疆维吾尔自治区自然科学基金(2019D01C173)。

摘　　要：目的探讨卡波西肉瘤(Kaposi's sarcoma,KS)相关疱疹病毒(KSHV)编码的微小RNAs(miR),如miR-K1-5p,对KS细胞周期的调控作用及其机制。方法该研究进行于2020年1—12月,人脐静脉内皮细胞购自美国ScienCell。生物信息学软件预测miR-K1-5p的靶基因。双萤光素酶试验验证miR-K1-5p的靶基因[细胞周期素依赖性激酶抑制剂4(CDKN4)]。将miR-K1-5p模拟物转染后,行蛋白质印迹法(Western blotting)和q-PCR检测,分析G1/S期相关基因CDKN4、细胞周期蛋白A/细胞周期素依赖性激酶2(Cyclin A/CDK2)、细胞周期蛋白D2/细胞周期素依赖性激酶4(Cyclin D2/CDK4)的表达。碘化丙啶(PI)染色检测miR-K1-5p对内皮细胞(HUVECs)周期的影响。结果miR-K1-5p靶向CDKN4(miR-K1-5p mimics+CDKN4-WT组、mimics NC+CDKN4-WT组、miR-K1-5p mimics+CDKN4-MUT组、mimics NC+CDKN4-MUT组萤光素酶活性分别为0.42±0.05、0.81±0.04、0.82±0.03、0.80±0.05),负性调控CDKN4的表达(P<0.001),正性调控Cyclin A/CDK2和CyclinD2/CDK4的表达(P<0.05)。此外,miR-K1-5p可以加速细胞周期进程,促进G1/S期转换[S+G2期/G1+S+G2期:mimics组(23.37±0.29)%比mimics NC组(15.00±0.75)%,G1期:mimics组(76.63±0.28)%比mimics NC组(85.00±0.76)%,S期:mimics组(15.34±0.36)%比mimics NC(8.45±0.20)%,G2期:mimics组(8.02±0.17)%比mimics NC组(6.55±0.80)%]。结论miR-K1-5p调控G1/S期相关基因的表达影响内皮细胞周期。Objective To explore the regulatory effect of Kaposi sarcoma-associated herpes virus(KSHV)-encoded micro-RNAs(miRNAs,miR)such as miR-K1-5p on KSHV cell cycle and the mechanism.Methods The experiment was conducted from January 2020 to December 2020.HUVECs cells were purchased from ScienCell in the United States.Bioinformatics software was used to predict the target gene of miR-K1-5p.Dual luciferase assay was used to verify the target gene[Cyclin-dependent kinase 4 inhibitor(CDKN4)]of miR-K1-5p.After miR-K1-5p mimics were transfected,Western blotting and q-PCR were used to analyze the expressions of G1/S phase related genes CDKN4,cell cycle protein A/Cyclin dependent kinase 2(Cyclin A/CDK2)and cell cycle protein D2/Cyclin dependent kinase 4(Cyclin D2/CDK4).Propidium Iodide(PI)staining was used to detect the effect of miR-K1-5p on HUVECs cell cycle.Results It proved that miR-K1-5p could target CDKN4 negatively regulate CDKN4 expression[The luciferase activities of miRK1-5p mimics+CDKN4-WT group,mimics NC+CDKN4-WT group,miR-K1-5p mimics+CDKN4-MUT group and mimics NC+CDKN4-MUT group were(0.42±0.05),(0.81±0.04),(0.82±0.03),and(0.80±0.05),respectively](P<0.001),and positively regulate Cyclin A/CDK2 and Cyclin D2/CDK4 expressions(P<0.05).In addition,it could accelerate cell cycle process and promote G1/S phase transition[S+G2 phase/G1+S+G2 phase:(23.37±0.29)%in mimics group vs.(15.00±0.75)%in mimics NC group;G1 phase:(76.63±0.28)%in mimics group vs.(85.00±0.76)%in mimics NC group;S phase:(15.34±0.36)%in mimics group vs.(8.45±0.20)%in mimics NC group;G2 phase:(8.02±0.17)%in mimics group vs.(6.55±0.80)%in mimics NC group].Conclusion miR-K1-5p affects the cycle of endothelial cells by regulating G1/S phase related genes.

关 键 词：内皮细胞 微小核糖核酸-K1-5p 细胞周期素依赖性激酶抑制剂4 细胞周期蛋白A/细胞周期素依赖性激酶2 细胞周期蛋白D2/细胞周期素依赖性激酶4 细胞周期 卡波西肉瘤 

分 类 号：R730.26[医药卫生—肿瘤]