本文關(guān)鍵詞：miR-711通過CD44調(diào)控胃癌細(xì)胞EMT　出處：《南華大學(xué)》2016年碩士論文　論文類型：學(xué)位論文

  更多相關(guān)文章： miR-711 CD44 EMT 胃癌細(xì)胞

【摘要】：目的:在課題組前期研究的基礎(chǔ)上,進(jìn)一步研究miR-711對(duì)CD44的調(diào)控作用及其與胃癌細(xì)胞發(fā)生EMT的相關(guān)性,初步揭示miR-711對(duì)胃癌侵襲轉(zhuǎn)移的調(diào)節(jié)機(jī)制。方法:1)使用TargetScan靶基因預(yù)測(cè)數(shù)據(jù)庫(kù)檢索miR-711與CD44 3'UTR的結(jié)合位點(diǎn),mi R-711 mimics與CD44 3'UTR質(zhì)粒共轉(zhuǎn)染293T細(xì)胞,熒光素酶報(bào)告基因?qū)嶒?yàn)檢測(cè)熒光素酶活性,驗(yàn)證miR-711對(duì)CD44的靶向調(diào)控作用。2)通過脂質(zhì)體轉(zhuǎn)染法將miR-711過表達(dá)(miR-711 mimics)、miR-711抑制表達(dá)(miR-711 inhibitors)、miRNA空質(zhì)粒(miRNA-NC)分別轉(zhuǎn)染人胃癌細(xì)胞株MGC-803及SGC-7901,以miRNA空質(zhì)粒轉(zhuǎn)染組作為陰性對(duì)照組,以空白轉(zhuǎn)染組作為空白對(duì)照組,轉(zhuǎn)染48小時(shí)后在熒光顯微鏡下觀察轉(zhuǎn)染效率,Western Blot實(shí)驗(yàn)檢測(cè)各組CD44蛋白的表達(dá)量。3)通過脂質(zhì)體轉(zhuǎn)染將CD44干擾質(zhì)粒(沉默組)、CD44空質(zhì)粒轉(zhuǎn)染人胃癌細(xì)胞株MGC-803及SGC-7901,以CD44空質(zhì)粒作為陰性對(duì)照組,以空白轉(zhuǎn)染組作為空白對(duì)照組,轉(zhuǎn)染48小時(shí)后在熒光顯微鏡下觀察轉(zhuǎn)染效率,Western Blot實(shí)驗(yàn)檢測(cè)各組CD44蛋白的表達(dá)量及EMT相關(guān)分子標(biāo)志蛋白E-cadherin、Vimentin的表達(dá)量。結(jié)果:1)生物信息學(xué)分析結(jié)果顯示miR-711與CD44 3'UTR之間存在結(jié)合位點(diǎn);熒光素酶報(bào)告基因?qū)嶒?yàn)結(jié)果顯示,與對(duì)照組相比,miR-711 mimics與CD44 3'UTR熒光素酶報(bào)告質(zhì)粒共轉(zhuǎn)染細(xì)胞后,相對(duì)熒光素酶活性減少,差異有統(tǒng)計(jì)學(xué)意義(p0.05)。2)人胃癌細(xì)胞株MGC-803、SGC-7901瞬時(shí)轉(zhuǎn)染miR-711過表達(dá)及抑制表達(dá)質(zhì)粒后,Western Blot結(jié)果均顯示,與miR-711 inhibitors組、陰性對(duì)照組及空白對(duì)照組相比,miR-711 mimics組CD44蛋白表達(dá)量減少,差異有統(tǒng)計(jì)學(xué)意義(p0.05);與陰性對(duì)照組及空白對(duì)照組相比,miR-711inhibitors組CD44蛋白表達(dá)量增多,差異有統(tǒng)計(jì)學(xué)意義(p0.05);陰性對(duì)照組與空白對(duì)照組相比,差異無(wú)統(tǒng)計(jì)學(xué)意義(p0.05)。3)人胃癌細(xì)胞株MGC-803、SGC-7901瞬時(shí)轉(zhuǎn)染CD44干擾質(zhì)粒后,Western Blot結(jié)果均顯示,與陰性組及空白組相比,沉默組CD44蛋白表達(dá)量減少,間質(zhì)標(biāo)志物Vimentin蛋白表達(dá)量減少,上皮標(biāo)志物E-cadherin蛋白表達(dá)量增多,差異有統(tǒng)計(jì)學(xué)意義(p0.05),陰性組與空白組相比,差異無(wú)統(tǒng)計(jì)學(xué)意義(p0.05)。結(jié)論:1、CD44為miR-711的靶基因之一;2、miR-711可通過下調(diào)CD44的表達(dá)抑制人胃癌細(xì)胞株MGC-803、SGC-7901發(fā)生EMT。
[Abstract]:Objective: Based on previous studies, further study the relationship between the role of miR-711 in regulation of CD44 and EMT in gastric cancer cells, revealed miR-711 regulating the invasion mechanism of metastasis of gastric cancer. Methods: 1) using the TargetScan target genes predicted binding sites of miR-711 and CD44 3'UTR database, MI R-711 mimics and CD44 3'UTR plasmids were transfected into 293T cells. Luciferase reporter assay of luciferase activity, verification of miR-711 targeted regulation of CD44.2) by liposome transfection over expression of miR-711 (miR-711 mimics), miR-711 (miR-711 inhibitors), inhibit the expression of miRNA empty plasmid (miRNA-NC) were transfected into human gastric cancer cell lines MGC-803 and SGC-7901, to miRNA empty plasmid group as negative control group, the blank transfection group as the control group, after 48 hours of transfection efficiency of transfection was observed under fluorescence microscopy, The expression of.3 protein was detected by CD44 Western Blot in each experiment) by liposome transfection of CD44 plasmid CD44 (silent group), empty plasmid were transfected into human gastric cancer cell lines MGC-803 and SGC-7901, with CD44 empty plasmid as a negative control group, the blank transfection group as the control group, after 48 hours of transfection efficiency of transfection was observed under fluorescence under the microscope, each symbol protein E-cadherin detection of CD44 protein expression and Western Blot experiment of EMT related molecules, the expression of Vimentin. Results: 1) analysis showed the presence of binding sites between miR-711 and CD44 3'UTR bioinformatics; luciferase reporter assay results showed that compared with the control group, miR-711 mimics and CD44 3'UTR luciferase reporter plasmid co transfection, luciferase activity decreased, the difference was statistically significant (P0.05).2) in human gastric cancer cell line MGC-803, SGC-7901 transfected miR-711 Overexpression and inhibition of the expression plasmid, Western Blot showed that, miR-711 and inhibitors group, compared with the negative control group and blank control group, miR-711 mimics group, CD44 protein expression decreased, the difference was statistically significant (P0.05); compared with the negative control group and blank control group, miR-711inhibitors group increased the protein expression of CD44, a statistically significant difference (P0.05); compared with the negative control group and blank control group, the difference was not statistically significant (P0.05).3) in human gastric cancer cell line MGC-803, SGC-7901 transient transfection of CD44 plasmid, Western Blot showed that, compared with the negative group and blank group, silent group CD44 protein expression decreased, mesenchymal marker Vimentin protein expression decreased epithelial marker expression of E-cadherin protein increased, the difference was statistically significant (P0.05), compared with the negative group and blank control group, the difference was not statistically significant (P0.05). Conclusion: 1 CD44 is one of the target genes of miR-711; 2, miR-711 can inhibit the human gastric cancer cell line MGC-803 by downregulating the expression of CD44, and SGC-7901 can occur EMT.

【學(xué)位授予單位】：南華大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2016
【分類號(hào)】：R735.2

【相似文獻(xiàn)】

相關(guān)期刊論文 前10條

1 王聰捷;劉秀香;謝書陽(yáng);宋曉東;呂長(zhǎng)俊;;紫杉醇對(duì)肺泡Ⅱ型上皮細(xì)胞EMT的干預(yù)作用[J];濱州醫(yī)學(xué)院學(xué)報(bào);2013年01期

2 寧亮,羅勇,賀大林;EMT在胚胎發(fā)育及腫瘤發(fā)生過程中的生物學(xué)作用[J];現(xiàn)代泌尿外科雜志;2005年05期

3 姜瑩;李曉燕;何慶南;譚愛斌;周頻;易著文;;白蛋白超載對(duì)人腎小管上皮細(xì)胞EMT及激素耐藥相關(guān)因素的影響[J];中國(guó)現(xiàn)代醫(yī)學(xué)雜志;2010年21期

4 林媛媛;婁遠(yuǎn)蕾;梁昌達(dá);謝淑佩;謝安;;人誘導(dǎo)多能干細(xì)胞自然分化過程中EMT變化研究[J];實(shí)驗(yàn)與檢驗(yàn)醫(yī)學(xué);2013年06期

5 陳進(jìn)宏;蔡端;馬保金;;腫瘤壞死因子相關(guān)凋亡誘導(dǎo)配體通過促進(jìn)死亡相關(guān)蛋白3表達(dá)抑制人胃癌細(xì)胞株的生長(zhǎng)[J];外科理論與實(shí)踐;2006年05期

6 周明利;盧先州;張樹友;;HIF-1α與EMT在腫瘤中的作用及意義研究進(jìn)展[J];中國(guó)普通外科雜志;2012年09期

7 李俊琴;李華;;脂肪酸合成酶在腫瘤EMT中作用機(jī)制的研究進(jìn)展[J];現(xiàn)代腫瘤醫(yī)學(xué);2014年08期

8 王少卿,王緒;奧曲肽對(duì)人胃癌細(xì)胞株抑制作用機(jī)制的實(shí)驗(yàn)研究[J];徐州醫(yī)學(xué)院學(xué)報(bào);2002年04期

9 馬銳,何紅梅,袁媛;大蒜素對(duì)人胃癌細(xì)胞株SGC-7901和BGC-823生長(zhǎng)的影響[J];腫瘤防治雜志;2005年04期

10 陳進(jìn)宏,馬保金,蔡端;重組人腫瘤壞死因子-α和5-氟尿嘧啶通過誘導(dǎo)死亡相關(guān)蛋白3表達(dá)抑制人胃癌細(xì)胞株的生長(zhǎng)[J];上海醫(yī)學(xué);2005年02期

相關(guān)會(huì)議論文 前10條

1 王沈玉;李敏;;人胃癌細(xì)胞株逆轉(zhuǎn)的實(shí)驗(yàn)研究[A];第九屆全國(guó)中西醫(yī)結(jié)合腫瘤學(xué)術(shù)研討會(huì)論文集[C];2002年

2 湯睿;朱正綱;瞿穎;李建芳;紀(jì)玉寶;劉炳亞;燕敏;林言箴;;溫?zé)狍w外對(duì)人胃癌細(xì)胞株生物學(xué)行為的影響[A];第四屆中國(guó)腫瘤學(xué)術(shù)大會(huì)暨第五屆海峽兩岸腫瘤學(xué)術(shù)會(huì)議論文集[C];2006年

3 徐向明;林建江;;EMT和腫瘤進(jìn)展[A];2011年浙江省肛腸外科學(xué)術(shù)大會(huì)暨結(jié)直腸肛門疾病診治新進(jìn)展學(xué)習(xí)班論文匯編[C];2011年

4 薛淑芳;許洪偉;;人胃癌細(xì)胞株MGC-803 FHIT基因轉(zhuǎn)染對(duì)阿霉素敏感性的影響[A];中華醫(yī)學(xué)會(huì)第七次全國(guó)消化病學(xué)術(shù)會(huì)議論文匯編（下冊(cè)）[C];2007年

5 胡曉;韓盛璽;;過氧化物酶體增殖物激活受體γ配體對(duì)胃癌細(xì)胞MCG-803生長(zhǎng)及基因表達(dá)的研究[A];中華醫(yī)學(xué)會(huì)第七次全國(guó)消化病學(xué)術(shù)會(huì)議論文匯編（下冊(cè)）[C];2007年

6 任陳;申洪;;上皮——間質(zhì)轉(zhuǎn)化(EMT)與腫瘤的研究進(jìn)展[A];中華醫(yī)學(xué)會(huì)病理學(xué)分會(huì)2009年學(xué)術(shù)年會(huì)論文匯編[C];2009年

7 舒曉亮;康凱;張穎瓊;王蘭芳;;L-精氨酸調(diào)控人胃癌細(xì)胞株SGC-7901凋亡及機(jī)制研究[A];第四屆全國(guó)中西醫(yī)結(jié)合營(yíng)養(yǎng)學(xué)術(shù)會(huì)議論文資料匯編[C];2013年

8 吳濤;徐惠綿;姜成鋼;于淼;賈蘭玲;;TGF-β_1SiRNA抑制人胃癌細(xì)胞株SGC-7901腹膜轉(zhuǎn)移的實(shí)驗(yàn)研究[A];第四屆中國(guó)腫瘤學(xué)術(shù)大會(huì)暨第五屆海峽兩岸腫瘤學(xué)術(shù)會(huì)議論文集[C];2006年

9 劉輝;左靜;郭建文;劉江惠;張杰英;左連富;;格爾德霉素對(duì)人胃癌細(xì)胞株MGC-803增殖與凋亡的影響[A];第五屆全國(guó)中醫(yī)藥免疫學(xué)術(shù)研討會(huì)——暨環(huán)境·免疫與腫瘤防治綜合交叉會(huì)議論文匯編[C];2009年

10 禹立霞;謝麗;魏嘉;沈潔;錢曉萍;劉寶瑞;;異甘草酸鎂對(duì)人胃癌細(xì)胞株藥物敏感性相關(guān)基因表達(dá)的影響[A];中國(guó)腫瘤內(nèi)科進(jìn)展 中國(guó)腫瘤醫(yī)師教育（2014）[C];2014年

相關(guān)重要報(bào)紙文章 前2條

1 記者 馬曉芳;揭秘華為架構(gòu)變動(dòng)：與EMT并行 董事會(huì)走向臺(tái)前[N];第一財(cái)經(jīng)日?qǐng)?bào);2011年

2 ;愛立信力助愛沙尼亞EMT推出商用HSDPA[N];人民郵電;2006年

，

本文編號(hào)：1394391