摘要
目的:通过研究mir-18b对滋养细胞HTR-8增殖、凋亡和凋亡相关蛋白的影响,了解mir-18b对人滋养细胞功能的调控作用,进一步明确mir-18b在子痫前期发生发展过程中的作用。方法:实验组通过化学方法合成mir-18b inhibitor,用脂质体2000包裹mir-18b inhibitor转入HTR-8细胞中,空白转染组为空白对照组。用Realtime-RT-PCR检测mir-18bmRNA水平的表达。应用流式细胞术检测细胞凋亡和增殖周期的变化。Western Blot检测P53、Bcl-2凋亡蛋白的表达。结果:Realtime-RT-PCR结果显示转染mir-18b inhibitor后mir-18b表达量与空白对照组相比表达量明显降低;细胞周期检测两组之间无明显差异;与空白对照组相比mir-18b inhibitor组细胞凋亡率增加三倍;Western Blot检测结果显示:转染了mir-18b inhibitor后P53的表达量增加,Bcl-2表达量减少。结论:研究结果显示,转染mir-18b inhibitor后细胞的凋亡率升高,P53表达量增加,Bcl-2的表达量减少。说明mir-18binhibitor可能通过调控P53与Bcl-2的表达增强了HTR-8细胞的凋亡能力。为研究PE的发病机理提供新的依据和线索。
Objective： IIn order to confirm the mir-18b plays an important role in the pathogenesis progress of preeclampsia, we have studied the effect ofmir-18b on the proliferation, apoptosis and relevant protein expressions of trophoblast cell （HTR-8）. Methods： The compound mir-18b inhibitor was synthesized by a chemical synthesis method, then wrapped with the Lipofectamine 2000 and transfected into HTR-8 cells. Blank lipofetamine group is the bland control group. Mir-18b was quantified using Realtime-reverse transcription-PCR. Flow cytometry was used to study the effect on cell cycle and cell apoptosis. The protein expression of P53, Bcl-2 were assayed by western blot. Results： With the treatment of mir-18b inhibitor, the expression of mir-18b in HTR-8 cell decreased. The flow cytometry result shows that cell cycle between the experiment group and control group was not obviously changed. Apoptosis of HTR-8 cell with the mir-18b inhibitor transfection increased in number threefold compared with control group. Protein expression of Bcl-2 in mir-18b inhibitor group decreased compared with control group, while the expression of P53 increased. Conclusion： The results show that mir-18b inhibitor can increase the apoptosis and the expression of P53, while decrease the protein expression of Bcl-2 in HTR-8 cell. It＇s indicate that mir-18b inhibitor enhance the ability of apoptosis through regulating P53 and Bcl-2. Aim to provide new evidence and clues for research the pathogenesis of PE.
出处
《现代生物医学进展》
CAS
2013年第12期2221-2224,共4页
Progress in Modern Biomedicine
基金
国家自然科学基金项目(30973208
31000660
81172636)
