| 张静,彭靖淇.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[J].安徽医药,2023,27(1):108-112. |
| 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 |
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| DOI：10.3969/j.issn.1009-6469.2023.01.024 |
| 中文关键词: 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 |
| 英文关键词: Endothelial cells miR-K1-5p CDKN4 Cyclin A/CDK2 Cyclin D2/CDK4 Cell cycle Kaposi′s sarcoma |
| 基金项目:新疆维吾尔自治区自然科学基金( 2019D01C173) |
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| 中文摘要: |
| 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-K15p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+CDKN4WT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 |
| 英文摘要: |
| 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 January2020 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 ofG1/S phase related genes CDKN4, cell cycle protein A/Cyclin dependent kinase 2 (Cyclin A/ CDK2) and cell cycle protein D2/Cyclindependent 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+CDKN4MUT 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. |
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