本文選題：微小R- + 小鼠胚胎成纖維細(xì)胞�。� 參考：《中國生物化學(xué)與分子生物學(xué)報(bào)》2016年11期

【摘要】：微小RNA(miRNAs)作為強(qiáng)大的基因表達(dá)調(diào)控子,廣泛參與多種生命過程,在細(xì)胞衰老進(jìn)程中的作用也日益受到關(guān)注。miR-223是一個典型的抑癌基因,可顯著抑制細(xì)胞增殖能力。miR-223與阿爾茨海默癥、心血管疾病以及類風(fēng)濕性關(guān)節(jié)炎等衰老相關(guān)疾病的發(fā)生發(fā)展密切相關(guān)。盡管如此,miR-223在細(xì)胞衰老進(jìn)程中的作用及其分子機(jī)制尚未見報(bào)道。本研究通過連續(xù)傳代建立了小鼠胚胎成纖維細(xì)胞(MEF細(xì)胞)的復(fù)制性衰老模型,并利用熒光定量qRT-PCR檢測發(fā)現(xiàn),miR-223在衰老MEF細(xì)胞中的表達(dá)水平顯著上調(diào)。隨后,通過轉(zhuǎn)染miR-223模擬物Agomir-223在MEF細(xì)胞中過表達(dá)miR-223。結(jié)果顯示,過表達(dá)miR-223可顯著促進(jìn)MEF細(xì)胞的衰老表型并抑制其增殖能力,而抑制miR-223的表達(dá)可延緩MEF細(xì)胞的復(fù)制性衰老進(jìn)程。進(jìn)一步利用生物信息學(xué)方法預(yù)測,獲得多個miR-223的候選衰老相關(guān)靶基因,包括Rasa1、Ddit4和Smad1等。然而,雙螢光素酶報(bào)告系統(tǒng)結(jié)果顯示,miR-223并不顯著影響其螢光強(qiáng)度,表明它們很可能并不是miR-223的下游靶基因。綜上所述,miR-223可顯著促進(jìn)MEF細(xì)胞復(fù)制性衰老,然而其調(diào)節(jié)細(xì)胞衰老進(jìn)程的分子機(jī)制依然有待深入研究。
[Abstract]:As a powerful gene expression regulator, miRNAs) is widely involved in a variety of life processes. The role of miR-223 in cell senescence is a typical tumor suppressor gene, which can significantly inhibit the proliferation of cells .miR-223 and Alzheimer's disease. The occurrence and development of aging related diseases such as cardiovascular disease and rheumatoid arthritis are closely related. However, the role of miR-223 in the process of cell senescence and its molecular mechanism have not been reported. In this study, the model of mouse embryonic fibroblast (MEF) was established by continuous subculture, and the expression level of miR-223 was significantly up-regulated in aging MEF cells by fluorescence quantitative qRT-PCR. Subsequently, miR-223 mimetic Agomir-223 was overexpressed in MEF cells. The results showed that overexpression of miR-223 could significantly promote the senescence phenotype and inhibit the proliferation of MEF cells, while inhibiting the expression of miR-223 could delay the process of replicative senescence of MEF cells. Further more, bioinformatics method was used to predict the candidate senescence related target genes of miR-223, including Rasa1 Ddit4 and Smad1. However, the results of the double luciferase reporting system showed that miR-223 did not significantly affect its fluorescence intensity, suggesting that they were probably not downstream target genes of miR-223. In conclusion, miR-223 can significantly promote the replicative senescence of MEF cells, but its molecular mechanism of regulating cellular senescence still needs to be further studied.
【作者單位】： 廣東醫(yī)科大學(xué)衰老研究所;廣東醫(yī)科大學(xué)生物化學(xué)與分子生物學(xué)研究所;廣東省醫(yī)學(xué)分子診斷重點(diǎn)實(shí)驗(yàn)室;
【基金】：國家自然科學(xué)基金(No.31600976,81671399,81170327) 廣東省自然科學(xué)基金(No.2014A030310027,2016A030313684) 廣東省醫(yī)學(xué)科研基金項(xiàng)目(No.A2015288) 廣東省攀登計(jì)劃專項(xiàng)資金(No.pdjh2016b0219)資助~~
【分類號】：R3416
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本文編號：2028809