lncRNA MALAT1靶向调控miR-214在睾丸缺血再灌注损伤中的作用及其机制  

Effect and mechanism of lncRNA IncRNA MALAT1 target regulating miR-214 on proliferation and apoptosis of GC-1 cells in testicular ischemia-reperfusion injury

作　　者：宁金卓[1] 卓栋 程帆[1] NING Jinzhuo;ZHUO Dong;CHENG Fan(Department of Urology,Renmin Hospital of Wuhan University,Wuhan 430000,China)

机构地区：[1]武汉大学人民医院泌尿外科,湖北武汉430000 [2]皖南医学院第一附属医院弋矶山医院泌尿外科,安徽芜湖241000

出　　处：《皖南医学院学报》2021年第1期13-17,共5页Journal of Wannan Medical College

基　　金：中央高校基本科研业务专项资金(413000227)。

摘　　要：目的:观察长链非编码RNA肺癌转移相关转录本1(IncRNA MALAT1)在睾丸缺血再灌注损伤(IRI)中对GC-1细胞增殖和凋亡的作用及其机制。方法:建立睾丸GC-1细胞缺氧复氧模型,qRT-PCR法检测不同复氧损伤时间点IncRNA MALAT1和微小RNA-214(miR-214)的表达变化;分别采用MTT实验、流式细胞术测定转染IncRNA MALAT1对GC-1细胞增殖和凋亡的影响;采用Western blot实验检测凋亡相关蛋白caspase-3和cytc蛋白表达;荧光素酶实验验证IncRNA MALAT1和miR-214的靶向关系,并采用qRT-PCR法检测转染IncRNA MALAT1后细胞中miR-214的表达变化。结果:IncRNA MALAT1随着复氧损伤时间的增加,其表达增高,在缺氧3 h/复氧24 h细胞上调最为明显,而miR-214的表达降低;过表达IncRNA MALAT1能显著抑制GC-1细胞增殖能力,促进细胞凋亡。沉默IncRNA MALAT1可使GC-1细胞的增殖能力增强,凋亡水平减弱;荧光素酶报告实验表明IncRNA MALAT1序列上有miR-214的结合位点。过表达IncRNA MALAT1抑制GC-1细胞miR-214表达,而沉默IncRNA MALAT1能促进miR-214表达。结论:IncRNA MALAT1可以通过靶向miR-214改变GC-1细胞的增殖和凋亡能力,从而影响睾丸IRI的发生发展。Objective:To investigate the effect and mechanism of long non-coding RNA IncRNA MALAT1(lncRNA IncRNA MALAT1)on the proliferation and apoptosis of GC-1 cells in testicular ischemia-reperfusion injury(IRI).Methods:The hypoxia reoxygenation model for testicular GC-1 cells was established,and qRT-PCR was used to examine the expression of IncRNA MALAT1 and miR-214 at different reoxygenation injury time points.MTT and flow cytometry were performed to detect the effect of transfected IncRNA MALAT1 on the proliferation and apoptosis of GC-1 cells.Western blot assay was used to measure the protein expression of caspase-3 and cytc,and luciferase reporter was used to verify the relationship between IncRNA MALAT1 and miR-214.qRT-PCR was performed to measure the expression level of miR-214 after IncRNA MALAT1 being transfected.Results:IncRNA MALAT1 expression was significantly up-regulated with prolonged reoxygenation injury,whereas miR-214 expression was notably decreased.Overexpression of IncRNA MALAT1 inhibited the proliferation of GC-1 cells and promoted the level of apoptosis,yet silence of IncRNA MALAT1 increased the proliferation of GC-1 cells and decreased the level of apoptosis.Luciferase reporter showed that miR-214 was the downstream target gene of IncRNA MALAT1.Overexpressed IncRNA MALAT1 led to significant inhibition of miR-214 expression,but silenced IncRNA MALAT1 promoted miR-214 expression of GC-1 cells.Conclusion:IncRNA MALAT1 could change the proliferation and apoptosis of GC-1 cells by targeting miR-214,thus affecting the development of testicular IRI.

关 键 词：睾丸缺血再灌注损伤 长链非编码RNA肺癌转移相关转录1 微小RNA-214 

分 类 号：R726.9[医药卫生—儿科]