摘要
目的研究kshv-mirT12VTp通过靶向调控细胞周期蛋白依赖性激酶抑制因子1 A(CDKN1A)影响卡波西肉瘤(KS)的细胞周期&方法生物信息学软件预测kshwmu-kl2NVp是否有CDKNlA的结合位点;双荧光素酶实验检测kshv-mir-k12-1-5p是否靶向结合CDKNlA;将kshv-mir-k12-1 Vp模拟物(mimic//抑制物(inhibitor)分别转染到卡波西肉瘤细胞株(SLK)中,Western blot和qRTTCR方法检测kshv-mir-k12-1-5p对CDKNlA的调控作用;PI法检测kshe-mU-k12-1-5p对KS细胞周期的影响&结果MiOee靶基因预测软件分析显示:kshwmiRkl2VTp与CDKNlA的3,非翻译区(3PTR)有结合位点;荧光素酶活性结果分析证实kshe-mi e-k12-1-5p的作用合位点(P<0.01);Western blot和qRTTCR结果显示:在蛋白水平和mRNA水平上,kshwmiRkl2NTp可显著降低CDKNlA的表达水平(P<0.01);PI实验结果显示:与其余各组比较,k/wmio kl2NTp mimics组PI值(51.43土0.75%)最高(P<0.01),能加速KS细胞周期进程;与其余各组比较,kshwmiRkl2N-5p inhibitor组PI值(3274±1.28)%最低#P<0.01),能减慢KS细胞周期进程&结论CDKNlA是kWiwmiRkl2NTp的靶基因,kshe-mi e-k12-1-5p通过靶CDKN1A的表达影响卡波西肉瘤的细胞周期.
Objective To investigate the impact of kshv-mir-k12-1-5p on Kaposi's sarcoma(KS)cell cyele by tar^-geting CDKN1A.Methods The bioinforamtics was used to predict whether the kshv-mir-k12-1-5p existed binding sites with CDKN1A.Dual luciferase assay was used to detect whether kshv-rmir-k12-1-5p can target CDKN1A.kshv-mir-k12-1-5p mimics/inhibitor was transfected into Kaposi's sarcoma cell line(SLK)respectively.Westem blot and qRT-PCR were used to detect the regulatory efect of kshv-mir-k12-1-5p on CDKN1A.PI method was used to detect the flfect of kshv-mir-k12-1-5p on KS cell cycle.Results Mirbase target gene prediction software analysis showed that kshv-mir-k12-1-5p had binding sites with 3'UTR of CDKN1 A;Luciferase activity results confirmned that kshv-mir-k12-1_5p can specifically act on binding sites(P<0.01);Western blot and qRT-PCR results showed that kshv-mir-k12-1-5p could significantly reduce the expression CDKN1 A at protein and mRNA levels(P<0.01);PI assay resuls showed that the mimics group had the highest PI value(51.43+0.75)%compared with other groups(P<0.01),which could accelerate the process of KS cell cyele.The inhibitor group had the lowest PI value(32.74土1.28)%compared with other groups(P<0.01),which could slow down the process of KS cell cyele.Conclusion CDKN1A is a target gene of kshv-mir-k12-1-5p,and kshv-mir-k12-1-5p may afect KS cell cycle by targeting and inhibiting CDKN1 A expression.
作者
张静
彭靖淇
吴秀娟
普雄明
Zhang Jing;Peng Jingqi;Wu Xiujuan(Dept of Pathology,The Pirst Department of Surgery,Ailiated Traditional Chinese Medicine Hospital,Xinjiang Medical University,Urumqi 830000;The Third Group,The Pirst Department of Surgery,Ailiated Traditional Chinese Medicine Hospital,Xinjiang Medical University,Urumqi 830000;Dept of Dermatology,Pepole's Hospial of Xinjiang,Uygur Auonomous Region,Urumqi 830000)
出处
《安徽医科大学学报》
CAS
北大核心
2019年第12期1923-1927,共5页
Acta Universitatis Medicinalis Anhui
基金
国家自然科学基金(编号:81660454)
新疆医科大学附属中医医院院级课题(编号:ZYY201702)
