LncRNA TTTY15通过调控miR-520a-3p表达对高糖诱导大鼠心肌细胞增殖和凋亡的影响  

Effects of LncRNA TTTY15 on the proliferation and apoptosis of high glucose-induced rat cardiomyocytes by regulating the expression of miR-520a-3p

作　　者：郝翠翠[1] 张青 HAO Cuicui;ZHANG Qing(Department of Endocrinology,Liaocheng Second People's Hospital,Liaocheng 252600,China;Department of Cardiology,Liaocheng Second People's Hospital,Liaocheng 252600,China)

机构地区：[1]聊城市第二人民医院内分泌科,山东聊城252600 [2]聊城市第二人民医院心内科,山东聊城252600

出　　处：《现代医学》2021年第1期46-52,共7页Modern Medical Journal

摘　　要：目的:探讨长链非编码RNA TTTY15(LncRNA TTTY15)对高糖诱导大鼠心肌细胞H9c2增殖和凋亡的影响及作用机制。方法:高糖诱导H9c2细胞后,实时荧光定量PCR(RT-qPCR)检测H9c2细胞中TTTY15和miR-520a-3p水平,四甲基噻唑蓝染色法(MTT)检测细胞增殖,流式细胞仪检测细胞凋亡,蛋白印迹(Western blotting)法检测细胞周期蛋白D1(cyclin D1)和活化的半胱天冬酶-3(C-caspase-3)蛋白水平。转染TTTY15小干扰RNA或miR-520a-3p模拟物至H9c2细胞,用上述相同方法观察下调TTTY15或上调miR-520a-3p对高糖诱导的H9c2细胞存活率、凋亡率以及cyclin D1和C-caspase-3蛋白表达的影响。双荧光素酶报告基因实验验证TTTY15与miR-520a-3p调控关系。结果:高糖促进了H9c2细胞中TTTY15表达、细胞凋亡率及C-caspase-3蛋白表达(P <0.05),降低了H9c2细胞存活率、miR-520a-3p表达及cyclin D1蛋白表达(P <0.05)。下调TTTY15或上调miR-520a-3p可提高高糖诱导的H9c2细胞存活率及cyclin D1蛋白水平(P <0.05),降低细胞凋亡率和C-caspase-3蛋白水平(P <0.05)。TTTY15在H9c2细胞中靶向负调控miR-520a-3p表达。下调miR-520a-3p可逆转下调TTTY15对H9c2细胞存活率、凋亡率及cyclin D1和C-caspase-3蛋白表达的影响。结论:TTTY15可能通过下调细胞中miR-520a-3p表达抑制高糖诱导的H9c2细胞增殖并促进细胞凋亡。Objective: To investigate the effects of long non-coding RNA( LncRNA) TTTY15 on high glucoseinduced rat cardiomyocyte H9 c2 proliferation and apoptosis and its mechanism. Methods: After H9 c2 cells were induced by high glucose,the levels of TTTY15 and miR-520 a-3 p in H9 c2 cells were detected by real-time fluorescence quantitative PCR( RT-qPCR);cell proliferation was detected by MTT assay;cells apoptosis was detected by flow cytometry;the protein levels of cyclin D1 and activated caspase-3( C-caspase-3) were detected by Western blotting. After TTTY15 small interfering RNA or miR-520 a-3 p mimic was transfected into H9 c2 cells,the same methods as described above were used to observe the effects of down-regulating TTTY15 or up-regulating miR-520 a-3 p on high glucose-induced H9 c2 cell survival rate,apoptosis rate and the protein expression levels of cyclin D1 and C-caspase-3. The dual luciferase reporter gene assay verified the regulatory relationship between TTTY15 and miR-520 a-3 p. Results: High glucose promoted the expression of TTTY15,apoptosis rate and the expression of C-caspase-3 protein in H9 c2 cells( P < 0. 05),but reduced the expression of miR-520 a-3 p,survival rate and the expression of cyclin D1 protein( P < 0. 05). Down-regulating TTTY15 or up-regulating miR-520 a-3 p increased high glucose-induced H9 c2 cell survival rate and the expression of cyclin D1 protein( P < 0. 05),while decreased apoptosis rate and the expression of C-caspase-3 protein( P < 0. 05). TTTY15 negatively regulated the expression of miR-520 a-3 p in H9 c2 cells. Down-regulating miR-520 a-3 p reversed the effects of down-regulating TTTY15 on H9 c2 cell survival rate,apoptosis rate and the expression levels of cyclin D1 and C-caspase-3 protein.Conclusion: TTTY15 inhibits high glucose-induced H9 c2 cell proliferation and promotes apoptosis by downregulating the expression of miR-520 a-3 p in cells.

关 键 词：TTTY15 miR-520a-3p 糖尿病 心肌细胞 增殖 凋亡 大鼠 

分 类 号：R-33[医药卫生] R363.1