本文選題：下咽腫瘤　切入點(diǎn)：細(xì)胞低氧　出處：《中華臨床醫(yī)師雜志(電子版)》2016年22期

【摘要】：目的觀察缺氧環(huán)境下咽癌Fadu細(xì)胞間質(zhì)化表型的改變與Postn蛋白表達(dá)的相關(guān)性。方法采用25μmol/L氯化鈷(Co Cl2)化學(xué)缺氧方法缺氧24 h,模擬Fadu缺氧環(huán)境。采用免疫熒光法檢測25μmol/L Co Cl2作用下,Postn蛋白的表達(dá),觀察上皮間質(zhì)化(EMT)相關(guān)蛋白的表達(dá)變化。通過構(gòu)建postn si RNA真核表達(dá)質(zhì)粒,瞬時轉(zhuǎn)染Fadu細(xì)胞,觀察Postn沉默對Fadu間質(zhì)化蛋白標(biāo)記表達(dá)的影響,采用劃痕實驗觀察Postn沉默對Fadu細(xì)胞遷移能力。結(jié)果免疫熒光結(jié)果顯示,25μmol/L Co Cl2作用細(xì)胞24 h,Postn表達(dá)明顯高于正常培養(yǎng)組,Vimentin、Snail和Twist蛋白表達(dá)也明顯高于對照組,表明低氧環(huán)境可促進(jìn)Fadu細(xì)胞增殖和提高細(xì)胞間質(zhì)化程度。采用Postn si RNA真核表達(dá)載體轉(zhuǎn)染Fadu細(xì)胞,免疫熒光和實時定量PCR結(jié)果顯示,構(gòu)建的si RNA表達(dá)載體的基因沉默效率為75.43%,轉(zhuǎn)染陽性的Fadu細(xì)胞Vimentin。劃痕實驗證實,Postn基因干擾轉(zhuǎn)染陽性組,細(xì)胞遷移率與轉(zhuǎn)染Control質(zhì)粒的對照組相比明顯降低(P0.01)。結(jié)論 Postn蛋白高表達(dá)與Fadu細(xì)胞低氧環(huán)境下的細(xì)胞增殖和間質(zhì)化有關(guān),沉默Postn基因能夠明顯抑制Fadu細(xì)胞的間質(zhì)化和遷移浸潤能力。
[Abstract]:Objective to observe the correlation between the interstitial phenotype of pharyngeal carcinoma Fadu cells and the expression of Postn protein under anoxic environment. Methods 25 渭 mol/L Cobalt Chloride Co Cl2) chemical anoxic method was used to anoxic for 24 h and simulated Fadu anoxic environment. Immunofluorescence assay was used to detect the expression of Postn protein. Expression of Postn protein in the presence of 25 渭 mol/L Co Cl2. To observe the expression of EMT-related proteins in epithelial mesenchymal cells, the eukaryotic expression plasmid of postn si RNA was constructed and transfected into Fadu cells, and the effect of Postn silencing on the expression of Fadu interstitial protein markers was observed. The effect of Postn silencing on the migration of Fadu cells was observed by scratch test. Results Immunofluorescence results showed that the expression of Postn and Twist protein was significantly higher than that in normal culture group after treated with 25 渭 mol/L Co Cl2 for 24 h. The results showed that hypoxic environment could promote the proliferation of Fadu cells and increase the degree of interstitial cells. Postn si RNA eukaryotic expression vector was used to transfect Fadu cells. The results of immunofluorescence and real-time quantitative PCR showed that, The gene silencing efficiency of the constructed si RNA expression vector was 75.43. The positive Fadu cells were confirmed by scratch test. Conclusion the high expression of Postn protein is related to the proliferation and interstitial of Fadu cells in hypoxic environment. Silencing Postn gene can significantly inhibit the interstitial and migration ability of Fadu cells.
【作者單位】： 吉林大學(xué)第一醫(yī)院耳鼻咽喉頭頸外科;
【基金】：吉林省衛(wèi)計委科技項目(2009Z021)
【分類號】：R739.63

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