【摘要】：目的探討miR-302a對葉酸缺乏小鼠胚胎干細(xì)胞(mESC)增殖和凋亡的影響。方法 mESC分為完全培養(yǎng)基組(對照組),無葉酸培養(yǎng)基組(無葉酸組),無葉酸培養(yǎng)基+miR-302amimic組(miR-302a組),通過RT-PCR進(jìn)行檢測miR-302a在完全培養(yǎng)基及無葉酸培養(yǎng)基中的表達(dá)。構(gòu)建miR-302amimic,后轉(zhuǎn)染到無葉酸培養(yǎng)基mESC中,采用MTT法檢測miR-302a mimic對mESC活力的影響,Annexin V-FITC/PI流式細(xì)胞術(shù)檢測miR-302amimic對mESC細(xì)胞凋亡的影響;流式細(xì)胞術(shù)檢測miR-302amimic對mESC細(xì)胞周期的影響;Western blot檢測及磷脂酰肌醇3羥激酶(PI3K)/蛋白激酶B(Akt)/哺乳動物雷帕霉素靶蛋白(mTOR)信號通路激活情況,以及下游分子細(xì)胞周期蛋白D1(CyclinD1)、p21、p27表達(dá)的影響。結(jié)果 RT-PCR證實(shí)無葉酸組中miR-302a表達(dá)量下降(P0.01)。與完全培養(yǎng)基比較,無葉酸培養(yǎng)基中,mESC細(xì)胞活力下降,細(xì)胞凋亡增加,細(xì)胞周期阻滯在G1期,Akt及mTOR磷酸化水平下降,CyclinD1表達(dá)下調(diào),p21及p27表達(dá)上調(diào),差異均具有統(tǒng)計(jì)學(xué)意義(P0.01)。與無葉酸組比較,miR-302a組中,mESC細(xì)胞活力上升,凋亡下降,G1期縮短,AKT及mTOR磷酸化水平提高,CyclinD1表達(dá)上調(diào),p21及p27表達(dá)下調(diào),差異均具有統(tǒng)計(jì)學(xué)意義(P0.01)。結(jié)論 miR-302a類似物能顯著抑制缺乏葉酸mESC凋亡,能促進(jìn)其增值,與PI3K/AKT/mTOR信號通路有關(guān)。
[Abstract]:Objective to investigate the effect of miR-302a on proliferation and apoptosis of embryonic stem cells (mESC) in folic acid deficient mice. Methods mESC was divided into complete medium group (control group), folic acid free medium group (folic acid free group) and folic acid free medium miR-302amimic group (miR-302a group). The expression of miR-302a in complete medium and folic acid free medium was detected by RT-PCR. MiR-302amimic, was constructed and transformed into folic acid free medium mESC. The effect of miR-302amimic on mESC activity was detected by MTT assay, the effect of miR-302amimic on apoptosis of mESC cells was detected by Annexin V-FITC/PI flow cytometry, and the effect of miR-302amimic on mESC cell cycle was detected by flow cytometry. Western blot was used to detect the activation of phosphatidylinositol 3-hydroxykinase (PI3K) / protein kinase B (Akt) / mammal rapamicin target protein (mTOR) signaling pathway, and the expression of downstream molecular cell cycle proteins D1 (CyclinD1), p21 and p27. Results RT-PCR confirmed that the expression of miR-302a in folic acid group decreased (P 0.01). Compared with the complete culture medium, the activity of mESC cells decreased, the apoptosis increased, the cell cycle arrest was in G 1 phase, the phosphorylation level of Akt and mTOR decreased, the expression of CyclinD1 was down-regulated, and the expression of p21 and p27 was up-regulated in folic acid-free medium, the difference was statistically significant (P 0.01). Compared with folic acid group, the activity of mESC cells was increased, apoptosis decreased, G 1 phase was shortened, AKT and mTOR phosphorylation level was increased, CyclinD1 expression was up-regulated, p21 and p27 expression was down-regulated in miR-302a group, the difference was statistically significant (P 0.01). Conclusion miR-302a analogues can significantly inhibit apoptosis of folic acid deficient mESC and promote its increment, which is related to PI3K/AKT/mTOR signaling pathway.
【作者單位】： 四川省醫(yī)學(xué)科學(xué)院/四川省人民醫(yī)院兒科;
【分類號】：R321

【相似文獻(xiàn)】

相關(guān)期刊論文 前10條

1 王青,尹嶺;葉酸治療高同型半胱氨酸血癥38例[J];中國老年學(xué)雜志;2002年03期

2 歐敬民;葉酸與腫瘤發(fā)生[J];國外醫(yī)學(xué).外科學(xué)分冊;2002年02期

3 周巖;馬靜;;葉酸抗炎作用研究進(jìn)展[J];國外醫(yī)學(xué)(衛(wèi)生學(xué)分冊);2009年03期

4 謝姣;王華;任廷遠(yuǎn);李元虎;;橙汁中葉酸的研究[J];食品與發(fā)酵科技;2009年05期

5 扶風(fēng);潘銘;小風(fēng);;多能手葉酸[J];食品與藥品;2011年12期

6 劉鵬;白亞娜;胡曉斌;劉磊;杜文琦;;孕產(chǎn)婦服用葉酸有效依從行為影響因素的logistic回歸分析[J];中國衛(wèi)生統(tǒng)計(jì);2012年01期

7 王新愛;;探討孕產(chǎn)婦葉酸的補(bǔ)充問題[J];中國醫(yī)藥指南;2012年35期

8 劉英華;陳瑛;;葉酸的檢測方法及臨床應(yīng)用研究進(jìn)展[J];中國婦幼保健;2013年01期

9 張麗群;;葉酸在臨床上的應(yīng)用進(jìn)展[J];中國現(xiàn)代藥物應(yīng)用;2013年06期

10 韓詠霞;;補(bǔ)充葉酸可降低心血管病的風(fēng)險[J];開卷有益(求醫(yī)問藥);2011年03期

相關(guān)會議論文 前10條

1 張繼軍;石曉麗;王南平;;葉酸功能研究進(jìn)展[A];營養(yǎng)與慢性病——中國營養(yǎng)學(xué)會第七屆理事會青年工作委員會第一次學(xué)術(shù)交流會議論文集[C];2010年

2 孫淑娜;桂永浩;宋后燕;王躍祥;鐘濤;;葉酸缺乏導(dǎo)致先天性心血管疾病的實(shí)驗(yàn)動物研究[A];中華醫(yī)學(xué)會第十四次全國兒科學(xué)術(shù)會議論文匯編[C];2006年

3 郝玲;;第七節(jié) 葉酸[A];中國營養(yǎng)學(xué)會DRIs修訂專家委員會第二次會議樣板稿匯編[C];2011年

4 郝玲;;第七節(jié) 葉酸[A];中國營養(yǎng)學(xué)會第二次膳食營養(yǎng)素參考攝入量研討會匯編[C];2011年

5 張，

本文編號：2511580