本文選題：細(xì)胞周期 + 細(xì)胞極性��； 參考：《重慶醫(yī)科大學(xué)學(xué)報(bào)》2017年11期

【摘要】：目的:觀察MARK2基因?qū)e La細(xì)胞極性與增殖的影響,探討其可能的機(jī)制。方法:脂質(zhì)體介導(dǎo)MARK2質(zhì)粒轉(zhuǎn)染He La細(xì)胞,G418抗性篩選獲得穩(wěn)定轉(zhuǎn)染的細(xì)胞株。TRITC-Phalloidin染色觀察細(xì)胞極性,平板克隆形成試驗(yàn)研究細(xì)胞貼壁生長(zhǎng)能力,流式細(xì)胞術(shù)分析細(xì)胞周期各時(shí)相細(xì)胞的比例,Western blot檢測(cè)總Rb蛋白及磷酸化Rb蛋白的水平。結(jié)果:成功構(gòu)建穩(wěn)定轉(zhuǎn)染MARK2的He La細(xì)胞系,與空質(zhì)粒對(duì)照組細(xì)胞相比,MARK2蛋白表達(dá)增強(qiáng)后He La細(xì)胞形態(tài)改變、上皮細(xì)胞極性恢復(fù);MARK2表達(dá)增強(qiáng)后He La細(xì)胞形成平板克隆數(shù)量明顯減少[(303.67±7.77)vs.(111.67±7.64),P=0.000],細(xì)胞周期G1期比例明顯增加[(48.82±0.84)%vs.(72.01±2.13)%,P=0.000]、S期比例減少[(36.60±0.58)%vs.(16.03±3.85)%,P=0.010],Rb蛋白的磷酸化水平明顯降低[(7.66±0.74)vs.(1.10±0.13),t=15.053,P=0.003]。結(jié)論:在He La細(xì)胞中,MARK2通過抑制Rb蛋白的磷酸化,使細(xì)胞周期停滯在G1期,抑制細(xì)胞增殖。MARK2能夠重建He La細(xì)胞上皮極性,協(xié)同調(diào)控細(xì)胞極性與細(xì)胞增殖兩條通路。
[Abstract]:Aim: to investigate the effect of MARK2 gene on the polarity and proliferation of He-La cells. Methods: the stable transfected cell line, TRITC-Phalloidin, was obtained by liposome mediated MARK2 plasmid transfection with He-La cell G418 resistance screening. The cell polarity was observed by TRITC-Phalloidin staining. Flow cytometry was used to analyze the proportion of cells in different phases of cell cycle. The levels of total RB protein and phosphorylated RB protein were detected by Western blot. Results: the He-La cell line transfected with MARK2 was successfully constructed. The morphological changes of He-La cells were observed after the expression of MARK2 protein was enhanced compared with that of the blank plasmid control cells. 涓婄毊緇嗚優(yōu)鏋佹，

本文編號(hào)：2036671