發(fā)布時(shí)間：2018-01-17 01:12

  本文關(guān)鍵詞：槲皮素對(duì)缺氧誘導(dǎo)的人肝癌細(xì)胞上皮間質(zhì)轉(zhuǎn)化影響的初步研究　出處：《安徽醫(yī)科大學(xué)》2017年碩士論文　論文類型：學(xué)位論文

  更多相關(guān)文章： 槲皮素 缺氧誘導(dǎo)因子 上皮間質(zhì)轉(zhuǎn)化 肝癌

【摘要】：目的:上皮間質(zhì)轉(zhuǎn)化(epithelial-mesenchymaltransition,EMT)是指在特定的生理和病理狀態(tài)下上皮細(xì)胞向間質(zhì)細(xì)胞轉(zhuǎn)化的現(xiàn)象。腫瘤細(xì)胞發(fā)生侵襲和遷移的主要機(jī)制之一即腫瘤細(xì)胞發(fā)生了上皮間質(zhì)轉(zhuǎn)化。誘導(dǎo)細(xì)胞發(fā)生上皮間質(zhì)轉(zhuǎn)化的因素有很多,缺氧也被認(rèn)為是其重要的誘導(dǎo)因素之一。槲皮素(Quercetin,Que)是廣泛存在于植物中的天然黃酮類化合物,具有抗炎、抗氧化和抗腫瘤等作用,已被證實(shí)可抑制多種腫瘤細(xì)胞的上皮間質(zhì)轉(zhuǎn)化進(jìn)而抑制腫瘤細(xì)胞的侵襲和遷移。但大量的實(shí)驗(yàn)研究?jī)H證明了槲皮素在常氧條件下對(duì)腫瘤細(xì)胞上皮間質(zhì)轉(zhuǎn)化的抑制效應(yīng),目前尚無(wú)研究報(bào)道槲皮素在缺氧條件下對(duì)腫瘤細(xì)胞上皮間質(zhì)轉(zhuǎn)化的影響。本研究通過(guò)體外細(xì)胞實(shí)驗(yàn)探討槲皮素對(duì)缺氧誘導(dǎo)的人肝癌Hep G2細(xì)胞上皮間質(zhì)轉(zhuǎn)化的影響及其可能的機(jī)制。方法:體外培養(yǎng)人肝癌Hep G2細(xì)胞,以100umol/L的氯化鈷(Cobaltous chloride,Co Cl2)作用于細(xì)胞建立缺氧模型,將對(duì)數(shù)生長(zhǎng)期的Hep G2細(xì)胞分為四組:對(duì)照組、Co Cl2組、Que組、Co Cl2+Que組,采用噻唑藍(lán)比色法(MTT)檢測(cè)不同濃度的槲皮素在常氧和缺氧條件下作用48h、72h后肝癌細(xì)胞的活力變化,并計(jì)算出相應(yīng)條件下槲皮素對(duì)肝癌細(xì)胞的半數(shù)抑制濃度;采用細(xì)胞劃痕實(shí)驗(yàn)檢測(cè)槲皮素作用后四組肝癌細(xì)胞遷移的距離并比較其差異;采用q PCR和Western blotting法分別檢測(cè)槲皮素作用后四組Hep G2細(xì)胞中缺氧誘導(dǎo)因子1α(hypoxia inducible factor-1alpha,HIF-1α)和上皮間質(zhì)轉(zhuǎn)化相關(guān)標(biāo)志因子Snail、E-cadherin和vimentin的m RNA及蛋白的表達(dá)水平。結(jié)果:MTT結(jié)果顯示槲皮素可顯著抑制Hep G2細(xì)胞的增殖,且在一定的濃度范圍內(nèi)其抑制作用存在劑量依賴性。槲皮素在常氧、缺氧條件下作用48h和72h的IC50值分別為69.25±3.02umol/L、56.78±1.99umol/L和48.76±2.63umol/L、41.07±2.51umol/L。細(xì)胞劃痕實(shí)驗(yàn)結(jié)果顯示槲皮素能明顯減緩常氧和缺氧條件下Hep G2細(xì)胞的遷移能力。q PCR實(shí)驗(yàn)結(jié)果顯示槲皮素可使Snail、vimentin的m RNA表達(dá)降低和E-cadherin的m RNA表達(dá)增加,而對(duì)HIF-1α的m RNA表達(dá)無(wú)影響。Western blotting實(shí)驗(yàn)結(jié)果顯示槲皮素可下調(diào)HIF-1α、Snail和vimentin的蛋白表達(dá)和上調(diào)E-cadherin的蛋白表達(dá)。結(jié)論:槲皮素能顯著抑制缺氧誘導(dǎo)的人肝癌Hep G2細(xì)胞的上皮間質(zhì)轉(zhuǎn)化,其機(jī)制可能與槲皮素抑制缺氧誘導(dǎo)的人肝癌細(xì)胞中HIF-1α蛋白的累積有關(guān)。
[Abstract]:Objective: to study the epithelial-mesenchymal transition. EMT). The transformation of epithelial cells into interstitial cells in certain physiological and pathological conditions. One of the main mechanisms of invasion and migration of tumor cells, I. E. epithelial interstitial transformation of tumor cells, induces the occurrence of epithelial cells. There are many factors involved in mesenchymal transformation. Quercetin Quercetinine (Quercetin) is a natural flavonoid widely found in plants and has anti-inflammatory properties. Antioxidation and anti-tumor effects. It has been proved that quercetin can inhibit the epithelial interstitial transformation of many tumor cells and thus inhibit the invasion and migration of tumor cells. However, a large number of experimental studies have only proved that quercetin inhibits the epithelial interstitial transformation of tumor cells under normoxic conditions. Effects. There is no study on the effect of quercetin on epithelial mesenchymal transformation of tumor cells under hypoxia. In this study, we studied the effects of quercetin on hypoxia induced Hep of human hepatocellular carcinoma in vitro. Effects of epithelial mesenchymal transformation in G2 cells and its possible mechanisms methods: human hepatoma Hep G2 cells were cultured in vitro. The model of hypoxia was established by the action of Cobaltous chloride Co Cl2 (100 umol / L) on the cells. The Hep G2 cells in logarithmic growth phase were divided into four groups: control group, Co Cl2 group, que group, Co Cl2 Que group. Thiazolyl blue colorimetric assay (MTT) was used to detect the changes of the viability of hepatoma cells treated with quercetin at different concentrations for 48 h or 72 h under normoxic and hypoxic conditions. The inhibition concentration of quercetin on hepatoma cells was calculated. Cell scratch assay was used to detect the migration distance of liver cancer cells in the four groups after quercetin treatment and the difference was compared. Q PCR and Western blotting methods were used to detect hypoxia inducible factor-1 偽 (HIF-1 偽) in four groups of Hep G2 cells after quercetin treatment. Hypoxia inducible factor-1alpha. HIF-1 偽 and Snail. Results the results showed that quercetin could significantly inhibit the proliferation of Hep G2 cells. The inhibitory effect of quercetin was in a dose-dependent manner in a dose-dependent manner. The IC50 values of quercetin exposed to normoxic oxygen, hypoxia for 48h and 72h were 69.25 鹵3.02 umol/ L, respectively. 56.78 鹵1.99umoll / L and 48.76 鹵2.63umol/ L respectively. 41.07 鹵2.51umol / L. Cell scratch test showed that quercetin could significantly decrease the migration ability of Hep G2 cells under normoxic and hypoxic conditions. Q. PCR results showed that quercetin could make Snail. The expression of m RNA in vimentin was decreased and the expression of m RNA in E-cadherin was increased. The results of Western blotting assay showed that quercetin could down-regulate HIF-1 偽. The protein expression of Snail and vimentin and up-regulation of E-cadherin protein were up-regulated. Conclusion: quercetin can significantly inhibit hypoxia induced Hep of human hepatocellular carcinoma. Epithelium mesenchymal transformation of G2 cells. The mechanism may be related to the inhibitory effect of quercetin on the accumulation of HIF-1 偽 protein in human hepatoma cells induced by hypoxia.
【學(xué)位授予單位】：安徽醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：R735.7

【參考文獻(xiàn)】

相關(guān)期刊論文 前1條

1 Ammar Natalwala;Robert Spychal;Chris Tselepis;;Epithelial-mesenchymal transition mediated tumourigenesis in the gastrointestinal tract[J];World Journal of Gastroenterology;2008年24期

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本文編號(hào)：1435640