过表达miR-200c对人髓核细胞凋亡的影响及其机制

发布时间：2018-04-23 08:54

本文选题：微小RNA-c + 人髓核细胞　；参考：《山东医药》2017年22期

【摘要】：目的观察过表达miR-200c对人髓核细胞(HNPCs)凋亡的影响,并探讨其可能的作用机制。方法体外培养HNPCs,将HNPCs分为观察组、阴性组和空白组,观察组转染hsa-miR-200c mimics,阴性组转染阴性对照mimics-NC,空白组未经任何干预。转染后24 h,采用real-time PCR法检测3组细胞miR-200c的表达,流式细胞仪测算细胞凋亡率,ELISA平板计数法检测Caspase3/7活性,Western blotting法检测3组细胞酪氨酸蛋白激酶受体B(Trk B)蛋白。结果观察组、阴性组和空白组细胞miR-200c相对表达量分别为8.11±0.21、1.07±0.04、0.99±0.05,细胞凋亡率分别为23.6%±3.1%、1.5%±0.2%、1.6%±0.1%,Caspase3/7活性分别为299.237±24.245、119.476±10.657、117.317±11.317,Trk B蛋白相对表达量分别为0.48±0.11、0.50±0.12、0.15±0.06,观察组与阴性组、空白组比较,P均0.01。结论过表达miR-200c能够促进HNPCs的凋亡,其机制可能与调控Trk B的表达有关。
[Abstract]:Objective to investigate the effect of overexpression of miR-200c on apoptosis of human nucleus pulposus cells (HNPCs) and its possible mechanism. Methods HNPCs were cultured in vitro. HNPCs was divided into observation group, negative group and blank group. The hsa-miR-200c mimicswere transfected into the observation group and the negative control group was transfected with mimics-NC.The blank group had no intervention. 24 hours after transfection, the expression of miR-200c was detected by real-time PCR assay. The apoptosis rate was measured by flow cytometry and the activity of Caspase3/7 was detected by Elisa plate counting method. The protein of tyrosine protein kinase receptor B(Trk B was detected by Western blotting assay. Results the relative expression of miR-200c and apoptosis were 8.11 卤0.21 卤1.07 卤0.04 卤0.99 卤0.05 and 299.237 卤24.24519.476 卤10.617117.317 卤11.3170.15 卤0.06 respectively in the observation group, negative group and blank group, and the apoptotic rate was 23.6% 卤3.21% 卤0.21.6% 卤0.1% 卤0.21% 卤0.1% in the blank group, respectively. The relative expression of the miR-200c protein was 0.48 卤0.110.50 卤0.120.15 卤0.06in the control group and the control group, respectively. The relative expression level of the miR-200c protein was 0.48 卤0.110.50 卤0.120.15 卤0.06in the control group compared with the control group, and the relative expression of the miR-200c protein was 299.237 卤24.24519.476 卤10.657117.317 卤11.317tTrk B protein in the control group. Conclusion overexpression of miR-200c can promote the apoptosis of HNPCs, and its mechanism may be related to the regulation of Trk B expression.
【作者单位】：广东医科大学附属佛山禅城医院;
【基金】：广东省佛山市卫生和计划生育局医学科研课题(20160148)
【分类号】：R681.53

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