天堂国产午夜亚洲专区-少妇人妻综合久久蜜臀-国产成人户外露出视频在线-国产91传媒一区二区三区

氡致BEAS-2B細(xì)胞凋亡中miRNA-34a與YY1的作用

發(fā)布時(shí)間:2018-11-28 16:12
【摘要】:目的:明確miR-34a負(fù)性調(diào)控YY1在氡致BEAS-2B細(xì)胞凋亡中的作用,為研究氡致支氣管上皮細(xì)胞凋亡的分子機(jī)制提供基礎(chǔ)資料。方法:對(duì)BEAS-2B細(xì)胞進(jìn)行常規(guī)培養(yǎng),胰酶消化后接種于Transwell培養(yǎng)皿,12h后將培養(yǎng)皿移至染毒裝置中進(jìn)行氡染毒,氡暴露濃度為20,000Bq/m3,染毒時(shí)間30min,連續(xù)染毒2次后作為染毒1代(Rn1)。分別染氡5代(Rn5)、10代(Rn10)、15代(Rn15)、20代(Rn20)后,流式細(xì)胞術(shù)檢測(cè)細(xì)胞凋亡率,Real-time PCR檢測(cè)miRNA-34a基因的表達(dá)水平,Western blot檢測(cè)YY1的蛋白表達(dá)水平。人工合成人源性miR-34a mimic、miR-34a inhibitor,分別瞬時(shí)轉(zhuǎn)入BEAS-2B細(xì)胞中,觀察對(duì)細(xì)胞凋亡率、凋亡相關(guān)蛋白Bcl-2、PARP-1和Bax表達(dá)的影響。生物信息學(xué)分析發(fā)現(xiàn)轉(zhuǎn)錄因子YY1的3’-UTR存在與miR-34a互補(bǔ)結(jié)合的位點(diǎn),且符合miRNA靶基因的條件。在BEAS-2B細(xì)胞中分別瞬時(shí)轉(zhuǎn)入miR-34a mimic、miR-34a inhibitor,觀察YY1表達(dá)水平的改變。構(gòu)建YY1過表達(dá)載體和沉默載體,觀察對(duì)細(xì)胞凋亡、Bcl-2、PARP-1和Bax表達(dá)的影響。結(jié)果:(1)細(xì)胞凋亡率隨氡染毒劑量的增加而升高,miRNA-34a表達(dá)水平隨氡染毒劑量的增加而增加,而YY1表達(dá)水平隨氡染毒劑量的增加而降低;(2)轉(zhuǎn)染miR-34a mimic組細(xì)胞凋亡率增加,Bcl-2和PARP-1表達(dá)下調(diào),Bax表達(dá)上調(diào);(3)轉(zhuǎn)染YY1過表達(dá)載體組細(xì)胞凋亡率減少,Bcl-2和PARP-1表達(dá)上調(diào),Bax表達(dá)下調(diào)。結(jié)論:本次試驗(yàn)結(jié)果提示,在氡致支氣管上皮細(xì)胞凋亡的過程中,mi R-34a通過負(fù)性調(diào)控YY1的表達(dá),使Bax表達(dá)上調(diào),Bcl-2和PARP-1表達(dá)下調(diào),從而最終促進(jìn)細(xì)胞的凋亡。但其確切的分子生物學(xué)機(jī)制還需更多的實(shí)驗(yàn)加以證實(shí)。
[Abstract]:Aim: to investigate the role of miR-34a in the negative regulation of YY1 in radon induced apoptosis of BEAS-2B cells, and to provide basic information for studying the molecular mechanism of radon induced apoptosis of bronchial epithelial cells. Methods: BEAS-2B cells were cultured routinely, trypsin was digested and inoculated into Transwell petri dish. After 12 hours, the culture dish was transferred to the device for radon exposure. The exposure concentration of radon was 20000Bq / m3, and the exposure time was 30 min. After continuous exposure for 2 times, it was used as the first generation (Rn1). After radon exposure to radon for 5 (Rn5), 10 (Rn10), 15 (Rn15) and 20 (Rn20), the apoptosis rate was detected by flow cytometry, and the expression level of miRNA-34a gene was detected by Real-time PCR., Western blot was used to detect the expression of YY1 protein. Synthetic human miR-34a mimic,miR-34a inhibitor, was transiently transferred into BEAS-2B cells to observe the effects on apoptosis rate, Bcl-2,PARP-1 and Bax expression of apoptosis-related proteins. Bioinformatics analysis showed that the 3'-UTR of transcription factor YY1 had complementary binding sites with miR-34a, and it was in line with the condition of miRNA target gene. The changes of YY1 expression were observed by transient transfer into miR-34a mimic,miR-34a inhibitor, in BEAS-2B cells. YY1 overexpression vector and silencing vector were constructed to observe the effect on apoptosis, Bcl-2,PARP-1 and Bax expression. Results: (1) the cell apoptosis rate increased with the increase of radon exposure dose, the expression of miRNA-34a increased with the increase of radon exposure dose, while the expression level of YY1 decreased with the increase of radon exposure dose. (2) the rate of apoptosis increased, the expression of Bcl-2 and PARP-1 down-regulated and the expression of Bax up-regulated in miR-34a mimic transfection group, (3) the rate of apoptosis decreased, the expression of Bcl-2 and PARP-1 increased, and the expression of Bax decreased in the group transfected with YY1 overexpression vector. Conclusion: in the process of radon induced apoptosis of bronchial epithelial cells, mi R-34a can up-regulate the expression of YY1, up-regulate the expression of Bax, and down-regulate the expression of Bcl-2 and PARP-1, thus promoting the apoptosis of cells. But more experiments are needed to confirm its exact molecular biological mechanism.
【學(xué)位授予單位】:蘇州大學(xué)
【學(xué)位級(jí)別】:碩士
【學(xué)位授予年份】:2015
【分類號(hào)】:R114

【參考文獻(xiàn)】

相關(guān)期刊論文 前2條

1 童建;;吸煙與氡暴露致肺癌的流行病學(xué)和生物學(xué)差異[J];環(huán)境與職業(yè)醫(yī)學(xué);2007年05期

2 胡錚;鐵軼;孫志賢;鄭曉飛;;電離輻射誘導(dǎo)microRNA研究進(jìn)展[J];軍事醫(yī)學(xué);2011年03期

相關(guān)碩士學(xué)位論文 前1條

1 孫秀錦;電離輻射誘發(fā)microRNA表達(dá)改變及其對(duì)輻射損傷調(diào)控機(jī)制[D];蘇州大學(xué);2011年

,

本文編號(hào):2363432

資料下載
論文發(fā)表

本文鏈接:http://sikaile.net/yixuelunwen/yufangyixuelunwen/2363432.html


Copyright(c)文論論文網(wǎng)All Rights Reserved | 網(wǎng)站地圖 |

版權(quán)申明:資料由用戶238b7***提供,本站僅收錄摘要或目錄,作者需要?jiǎng)h除請(qǐng)E-mail郵箱bigeng88@qq.com
亚洲av秘片一区二区三区| 开心久久综合激情五月天| 香蕉久久夜色精品国产尤物| 午夜福利直播在线视频| 国产中文字幕一二三区| 大香蕉网国产在线观看av| 91日韩欧美国产视频| 免费国产成人性生活生活片| 伊人久久青草地综合婷婷| 国产精品超碰在线观看| 成人午夜视频在线播放| 好吊妞视频只有这里有精品| 亚洲av秘片一区二区三区| 国产精品一区二区三区日韩av | 欧美日韩免费观看视频| 婷婷色网视频在线播放| 99精品人妻少妇一区二区人人妻| 国产免费成人激情视频| 国产欧美一区二区另类精品| 欧美多人疯狂性战派对| 国产av天堂一区二区三区粉嫩| 91精品视频免费播放| 人妻精品一区二区三区视频免精| 日韩精品一区二区一牛| 欧美人与动牲交a精品| 日韩高清中文字幕亚洲| 欧美日韩精品综合在线| 丰满人妻一二区二区三区av| 大香蕉久草网一区二区三区| 精品一区二区三区乱码中文| 日韩日韩欧美国产精品| 亚洲av首页免费在线观看| 国产精品免费视频久久| 成人欧美一区二区三区视频| 婷婷色国产精品视频一区| 亚洲熟妇中文字幕五十路| 日韩精品一区二区一牛| 熟女免费视频一区二区| 久久这里只精品免费福利| 粗暴蹂躏中文一区二区三区| 日韩高清毛片免费观看|