MiR-23b-3p靶向調(diào)控EBF3影響食管癌生長(zhǎng)的機(jī)制研究
發(fā)布時(shí)間:2021-11-14 04:52
研究背景食管癌(Esophageal Cancer)是消化道最常見(jiàn)的惡性腫瘤之一,其全球發(fā)病率和死亡率分別占第8位和第6位;主要有腺癌(Esophageal Adenocarcinoma,EAC)和鱗狀細(xì)胞癌(Esophageal Squamous Cell Carcinoma,ESCC)兩種病理組織類型。在我國(guó),ESCC占到90%以上,手術(shù)+放化療綜合治療后的5年生存率仍非常低。因此,深入闡明食管癌發(fā)生發(fā)展的分子機(jī)制,對(duì)早期診斷、發(fā)現(xiàn)新的治療靶點(diǎn),改善預(yù)后具有重要意義。MicroRNA(miRNAs)是一類真核生物內(nèi)源性非編碼小分子RNAs,能負(fù)性調(diào)控下游靶基因,參與多種腫瘤的發(fā)生發(fā)展。MiR-23b-3p在胃癌、結(jié)直腸癌、軟骨肉瘤、甲狀腺濾泡癌、乳腺癌、神經(jīng)膠質(zhì)瘤、急性髓系白血病等多種腫瘤中表達(dá)異常,是非小細(xì)胞肺癌的非侵入性預(yù)后標(biāo)志物,與大腸癌耐藥密切相關(guān)。但miR-23b-3p在食管癌中的表達(dá)與作用機(jī)制未見(jiàn)報(bào)道。研究目的探討miR-23b-3p及其靶基因早期B細(xì)胞因子3(early B-cellfactor 3,EBF3)在食管癌發(fā)生發(fā)展中的作用及其相關(guān)機(jī)制。研究方法1.實(shí)時(shí)熒光...
【文章來(lái)源】:南方醫(yī)科大學(xué)廣東省
【文章頁(yè)數(shù)】:117 頁(yè)
【學(xué)位級(jí)別】:博士
【部分圖文】:
圖1-2MTT法檢測(cè)細(xì)胞增殖
A?B??圖1-2MTT法檢測(cè)細(xì)胞增殖。TE1?(A圖)和TE10?(B圖)細(xì)胞株轉(zhuǎn)染anti-miR-NC??和anti-miR-23b-3p后在第24h,48h,?72h檢測(cè)細(xì)胞增殖情況(與對(duì)照組相比,*P<0.05,??**P<0.01)??Figurel-2?MTT?assay?was?performed?for?cell?proliferation.?TE1?(FigureA)?and?TE10??(FigureB)?cells?transfected?with?anti-miR-NC?or?anti-miR-23b-3p?were?harvested?and?counted??after?24h,?48h.?72h?(Compared?with?the?control?group,?*P?<?0.05,?**P?<?0.01)??3.1.3下調(diào)miR-23b-3p表達(dá)能抑制ESCC細(xì)胞侵襲和轉(zhuǎn)移??MTT實(shí)驗(yàn)證實(shí)抑制miR-23b-3p表達(dá)能顯著抑制ESCC細(xì)胞增殖,那么??miR-23b-3p能影響ESCC細(xì)胞的體外侵襲和遷移能力嗎??Transwell實(shí)驗(yàn)結(jié)果顯??示,在轉(zhuǎn)染miR-23b-3p抑制劑后,TE1和TE10細(xì)胞株中發(fā)生侵襲和遷移的細(xì)??胞數(shù)量明顯少于轉(zhuǎn)染anti-miR-NC的對(duì)照組,差異具有統(tǒng)計(jì)學(xué)意義(圖1-3?A、??B,圖1-4?A、B),說(shuō)明腫瘤細(xì)胞的侵襲轉(zhuǎn)移能力受到了抑制。結(jié)合MTT實(shí)驗(yàn)??結(jié)果
?TE10??B??圖1-3下調(diào)miR-23b-3p表達(dá)對(duì)細(xì)胞侵襲能力的影響。TE1和TE10細(xì)胞株轉(zhuǎn)染??anti-miR-NC和anti-miR-23b-3p?24h后,進(jìn)行Transwell小室侵襲實(shí)驗(yàn)。隨機(jī)挑選5個(gè)視野,??放大100倍進(jìn)行觀察,并對(duì)發(fā)生侵襲轉(zhuǎn)移的細(xì)胞進(jìn)行計(jì)數(shù)。A圖所示為鏡下觀,B圖所示??為發(fā)生侵襲轉(zhuǎn)移的細(xì)胞計(jì)數(shù)(與對(duì)照組相比,叩<0.05,?**P<?0.01)??Figurel-3?Effect?of?down-regulated?miR-23b-3p?on?cell?invasion.?Transwell?invasion?assay??was?carried?out?in?ESCC?cell?lines?transfected?with?anti-miR-NC?and?anti-iniR-23b-3p.?Cells?in??five?random?fields?of?view?at?10〇x?magnification?were?observed?and?the?migrated?cells?were??counted.?(Compared?with?the?control?group,?*P<0.05,?**P?<0.01)??14??
【參考文獻(xiàn)】:
期刊論文
[1]Genomic characterization of esophageal squamous cell carcinoma: Insights from next-generation sequencing[J]. Yasushi Sasaki,Miyuki Tamura,Ryota Koyama,Takafumi Nakagaki,Yasushi Adachi,Takashi Tokino. World Journal of Gastroenterology. 2016(07)
[2]微小RNA-21通過(guò)調(diào)控上皮-間充質(zhì)轉(zhuǎn)化對(duì)膽管癌細(xì)胞侵襲與轉(zhuǎn)移的影響[J]. 劉振,黃強(qiáng),劉臣海,林先盛,謝放,朱成林,邵峰. 中華實(shí)驗(yàn)外科雜志. 2014 (11)
[3]唾液與血漿微小RNA-21對(duì)早期食管癌的診斷價(jià)值[J]. 葉敏華,葉鵬輝,張偉珠,饒珈琦,謝子鈞. 南方醫(yī)科大學(xué)學(xué)報(bào). 2014(06)
[4]唾液中miRNA-144可作為早期診斷食道癌的基因標(biāo)志物[J]. 吳偉東,侯文進(jìn),吳哲凡,王躍軍,易永盛,林葦嘉. 南方醫(yī)科大學(xué)學(xué)報(bào). 2013(12)
[5]肺癌A549細(xì)胞上皮-間質(zhì)轉(zhuǎn)化及其對(duì)微小RNA表達(dá)的影響[J]. 王海兵,張慧君,蘇晉梅,周文勇,王和勇,陳曉峰. 中華腫瘤雜志. 2011 (08)
[6]MicroRNAs, development of Barrett’s esophagus, and progression to esophageal adenocarcinoma[J]. Cameron M Smith,David I Watson,Michael Z Michael,Damian J Hussey. World Journal of Gastroenterology. 2010(05)
[7]早期B細(xì)胞因子3在肝癌中的表達(dá)及對(duì)HepG2細(xì)胞株作用的研究[J]. 王躍國(guó),鞠少卿,王惠民. 中華微生物學(xué)和免疫學(xué)雜志. 2009 (09)
本文編號(hào):3493998
【文章來(lái)源】:南方醫(yī)科大學(xué)廣東省
【文章頁(yè)數(shù)】:117 頁(yè)
【學(xué)位級(jí)別】:博士
【部分圖文】:
圖1-2MTT法檢測(cè)細(xì)胞增殖
A?B??圖1-2MTT法檢測(cè)細(xì)胞增殖。TE1?(A圖)和TE10?(B圖)細(xì)胞株轉(zhuǎn)染anti-miR-NC??和anti-miR-23b-3p后在第24h,48h,?72h檢測(cè)細(xì)胞增殖情況(與對(duì)照組相比,*P<0.05,??**P<0.01)??Figurel-2?MTT?assay?was?performed?for?cell?proliferation.?TE1?(FigureA)?and?TE10??(FigureB)?cells?transfected?with?anti-miR-NC?or?anti-miR-23b-3p?were?harvested?and?counted??after?24h,?48h.?72h?(Compared?with?the?control?group,?*P?<?0.05,?**P?<?0.01)??3.1.3下調(diào)miR-23b-3p表達(dá)能抑制ESCC細(xì)胞侵襲和轉(zhuǎn)移??MTT實(shí)驗(yàn)證實(shí)抑制miR-23b-3p表達(dá)能顯著抑制ESCC細(xì)胞增殖,那么??miR-23b-3p能影響ESCC細(xì)胞的體外侵襲和遷移能力嗎??Transwell實(shí)驗(yàn)結(jié)果顯??示,在轉(zhuǎn)染miR-23b-3p抑制劑后,TE1和TE10細(xì)胞株中發(fā)生侵襲和遷移的細(xì)??胞數(shù)量明顯少于轉(zhuǎn)染anti-miR-NC的對(duì)照組,差異具有統(tǒng)計(jì)學(xué)意義(圖1-3?A、??B,圖1-4?A、B),說(shuō)明腫瘤細(xì)胞的侵襲轉(zhuǎn)移能力受到了抑制。結(jié)合MTT實(shí)驗(yàn)??結(jié)果
?TE10??B??圖1-3下調(diào)miR-23b-3p表達(dá)對(duì)細(xì)胞侵襲能力的影響。TE1和TE10細(xì)胞株轉(zhuǎn)染??anti-miR-NC和anti-miR-23b-3p?24h后,進(jìn)行Transwell小室侵襲實(shí)驗(yàn)。隨機(jī)挑選5個(gè)視野,??放大100倍進(jìn)行觀察,并對(duì)發(fā)生侵襲轉(zhuǎn)移的細(xì)胞進(jìn)行計(jì)數(shù)。A圖所示為鏡下觀,B圖所示??為發(fā)生侵襲轉(zhuǎn)移的細(xì)胞計(jì)數(shù)(與對(duì)照組相比,叩<0.05,?**P<?0.01)??Figurel-3?Effect?of?down-regulated?miR-23b-3p?on?cell?invasion.?Transwell?invasion?assay??was?carried?out?in?ESCC?cell?lines?transfected?with?anti-miR-NC?and?anti-iniR-23b-3p.?Cells?in??five?random?fields?of?view?at?10〇x?magnification?were?observed?and?the?migrated?cells?were??counted.?(Compared?with?the?control?group,?*P<0.05,?**P?<0.01)??14??
【參考文獻(xiàn)】:
期刊論文
[1]Genomic characterization of esophageal squamous cell carcinoma: Insights from next-generation sequencing[J]. Yasushi Sasaki,Miyuki Tamura,Ryota Koyama,Takafumi Nakagaki,Yasushi Adachi,Takashi Tokino. World Journal of Gastroenterology. 2016(07)
[2]微小RNA-21通過(guò)調(diào)控上皮-間充質(zhì)轉(zhuǎn)化對(duì)膽管癌細(xì)胞侵襲與轉(zhuǎn)移的影響[J]. 劉振,黃強(qiáng),劉臣海,林先盛,謝放,朱成林,邵峰. 中華實(shí)驗(yàn)外科雜志. 2014 (11)
[3]唾液與血漿微小RNA-21對(duì)早期食管癌的診斷價(jià)值[J]. 葉敏華,葉鵬輝,張偉珠,饒珈琦,謝子鈞. 南方醫(yī)科大學(xué)學(xué)報(bào). 2014(06)
[4]唾液中miRNA-144可作為早期診斷食道癌的基因標(biāo)志物[J]. 吳偉東,侯文進(jìn),吳哲凡,王躍軍,易永盛,林葦嘉. 南方醫(yī)科大學(xué)學(xué)報(bào). 2013(12)
[5]肺癌A549細(xì)胞上皮-間質(zhì)轉(zhuǎn)化及其對(duì)微小RNA表達(dá)的影響[J]. 王海兵,張慧君,蘇晉梅,周文勇,王和勇,陳曉峰. 中華腫瘤雜志. 2011 (08)
[6]MicroRNAs, development of Barrett’s esophagus, and progression to esophageal adenocarcinoma[J]. Cameron M Smith,David I Watson,Michael Z Michael,Damian J Hussey. World Journal of Gastroenterology. 2010(05)
[7]早期B細(xì)胞因子3在肝癌中的表達(dá)及對(duì)HepG2細(xì)胞株作用的研究[J]. 王躍國(guó),鞠少卿,王惠民. 中華微生物學(xué)和免疫學(xué)雜志. 2009 (09)
本文編號(hào):3493998
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