【摘要】：目的:探討轉(zhuǎn)化生長(zhǎng)因子(transforming growth factor-β,TGF-β1)在食管癌中的作用及其對(duì)上皮間質(zhì)轉(zhuǎn)化(Epithelial-Mesenchymal Transition,EMT)的調(diào)控。方法:利用TGF-β1受體抑制劑LY2109761處理食管癌Eca109細(xì)胞,根據(jù)濃度梯度(0、0.1、1、10μm/ml)分為四組;慢病毒轉(zhuǎn)染TGF-β1干擾RNA(siRNA)處理食管癌Eca109細(xì)胞,分為3組:實(shí)驗(yàn)組(病毒穩(wěn)定TGF-β1 si RNA轉(zhuǎn)染Eca109細(xì)胞)、陰性對(duì)照組(轉(zhuǎn)染無(wú)關(guān)序列Eca109細(xì)胞)、空白對(duì)照組(正常未經(jīng)任何處理的Eca109細(xì)胞),采用熒光定量PCR(Quantitative Realtime-PCR,qRT-PCR)技術(shù)檢測(cè)TGF-β1及上皮間質(zhì)轉(zhuǎn)化(EMT)相關(guān)指標(biāo)E-鈣粘蛋白(E-cadherin)、波形蛋白(Vimentin)、鋅指轉(zhuǎn)錄因子(Snail)在mRNA水平表達(dá)情況;Western blot技術(shù)檢測(cè)E-cadherin、Vimentin蛋白水平的表達(dá);同時(shí)通過(guò)細(xì)胞增殖實(shí)驗(yàn)(MTT)、細(xì)胞劃痕實(shí)驗(yàn)及Transwell法分別檢測(cè)細(xì)胞增殖、遷移、侵襲的變化情況。結(jié)果:通過(guò)TGF-β1受體抑制劑(0、0.1、1、10μm/ml)處理Eca109細(xì)胞后,在mRNA水平TGF-β1表達(dá)逐漸降低,E-cadherin表達(dá)逐漸升高,Vimentin、Snail表達(dá)逐漸降低(P0.05);在蛋白水平,E-cadherin表達(dá)逐漸升高,Vimentin表達(dá)逐漸降低(P0.05);此外,細(xì)胞增殖和遷移能力逐漸降低(P0.05)。通過(guò)慢病毒轉(zhuǎn)染TGF-β1 si RNA后發(fā)現(xiàn),實(shí)驗(yàn)組與空白對(duì)照組和陰性對(duì)照組相比,TGF-β1 mRNA表達(dá)降低(P0.05),E-cadherin表達(dá)增高,Vimentin表達(dá)降低;同時(shí)實(shí)驗(yàn)組細(xì)胞增殖、遷移和侵襲能力均降低(P0.05)。結(jié)論:TGF-β1可能通過(guò)上皮間質(zhì)轉(zhuǎn)化促進(jìn)食管癌細(xì)胞的增殖、遷移和侵襲。
[Abstract]:Aim: to investigate the role of transforming growth factor (transforming growth factor- 尾 1 (TGF- 尾 1) in esophageal carcinoma and its regulation on Epithelial-Mesenchymal transition. Methods: esophageal carcinoma Eca109 cells were treated with TGF- 尾 1 receptor inhibitor (LY2109761) and divided into four groups according to the concentration gradient (0 0. 1 渭 m/ml), lentivirus transfected with TGF- 尾 1 interfered with RNA (siRNA) to treat esophageal cancer Eca109 cells. TGF- 尾 1 was divided into three groups: experimental group (viral stable TGF- 尾 1si RNA transfected Eca109 cells), negative control group (transfection of unrelated Eca109 cells) and blank control group (normal untreated Eca109 cells). TGF- 尾 1 was detected by fluorescence quantitative PCR (Quantitative real time PCR qRT-PCR. Ecadherin (E-cadherin) and (Vimentin), zinc finger transcription factor (Snail) (Snail) were detected by Western blot. At the same time, the cell proliferation, migration and invasion were detected by (MTT), cell scratch assay and Transwell assay. Results: after treated with TGF- 尾 1 receptor inhibitor (0 0. 1 m/ml), the expression of TGF- 尾 1 decreased gradually at the mRNA level and decreased gradually at the mRNA level (P0.05), while at the protein level, the expression of E-cadherin decreased gradually (P0.05), and the expression of Vimentin decreased gradually at the protein level (P0.05). The ability of cell proliferation and migration decreased gradually (P0.05). After lentivirus transfection of TGF- 尾 1si RNA, it was found that the expression of TGF- 尾 1 mRNA in the experimental group was lower than that in the blank control group and the negative control group (P0.05), while the expression of Vimentin in the experimental group was lower than that in the control group (P0.05), and the ability of cell proliferation, migration and invasion in the experimental group was decreased (P0.05). Conclusion TGF- 尾 1 may promote the proliferation, migration and invasion of esophageal carcinoma cells through epithelial interstitial transformation.
【學(xué)位授予單位】：新疆醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2016
【分類(lèi)號(hào)】：R735.1

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