本文選題：胃癌 + 乏氧誘導(dǎo)因子-1α　； 參考：《新疆醫(yī)科大學(xué)》2017年碩士論文

【摘要】：目的:觀察不同缺氧條件下胃腺癌細(xì)胞系BGC-823中HIF-1α、HIF-2α、HIF-1β和VEGF的表達(dá),探討三種HIF因子在低氧條件下調(diào)控胃癌血管生成的作用差異。方法:分別用150、200、250、275μmol/L Co Cl2模擬腫瘤內(nèi)部缺氧,缺氧時間分別為:2h、4h、8h、12h、24h、48h、72h。采用實(shí)時定量聚合酶鏈反應(yīng)(Real-time PCR)和Western blot法檢測HIF-1α、HIF-2α、HIF-1β和VEGF的mRNA及蛋白表達(dá)情況。進(jìn)一步運(yùn)用RNA干擾(RNA interference,RNAi)技術(shù)采用小干擾RNA(small interfering RNA,siRNA)沉默BGC-823細(xì)胞,比較三者沉默VEGF的效果。結(jié)果:1.與常氧時比較,缺氧后HIF-1α、HIF-2α、VEGF的m RNA及蛋白表達(dá)均隨CoCl2濃度增加、時間延長而增加,但三者變化趨勢不一致,HIF-1α在缺氧早期迅速升高,之后快速下降;HIF-2α和VEGF則在缺氧晚期表達(dá)最高,之后保持穩(wěn)定。HIF-1βmRNA和蛋白在常氧或缺氧下表達(dá)均無明顯變化(P0.05)。2.細(xì)胞分別特異性轉(zhuǎn)染HIF-1α、HIF-2α和HIF-1βsiRNA后均能有效沉默各自基因的表達(dá),同時VEGF的表達(dá)水平也隨之降低,且隨HIF-2α的下調(diào)幅度最顯著(P0.05)。結(jié)論:1.BGC-823胃癌細(xì)胞中HIF-1α、HIF-2α和HIF-1β對乏氧刺激表現(xiàn)出不同的應(yīng)答方式,HIF-1α和HIF-2α可能是胃癌適應(yīng)乏氧環(huán)境的主要缺氧誘導(dǎo)因子;2.RNA干擾能抑制三種HIF基因的表達(dá),并不同程度減少下游VEGF的表達(dá),相較于HIF-1α和HIF-1β沉默途徑,切斷HIF-2α可能會更有效阻止胃癌血管生成。
[Abstract]:Objective: to observe the expression of HIF-1 偽 -HIF-2 偽 -hIF-1 尾 and VEGF in gastric adenocarcinoma cell line BGC-823 under different hypoxia conditions, and to explore the difference of three HIF factors in regulating angiogenesis of gastric cancer under hypoxia. Methods: 150200250275 渭 mol/L Co Cl2 was used to simulate the anoxia in the tumor. The anoxia time of the tumor was 12: 2, 4 h, 8 h, 12 h, 24 h, 48 h, 72 h, respectively. Real-time polymerase chain reaction (Real-time PCR) and Western blot were used to detect the expression of mRNA and protein in HIF-1 偽 -HIF-2 偽 HIF-1 尾 and VEGF. RNA interference siRNAs were further used to silence BGC-823 cells. The effect of VEGF silencing was compared among the three groups. The result is 1: 1. The expression of m RNA and protein of HIF-1 偽 -HIF-2 偽 -VEGF increased with the increase of CoCl2 concentration and time after hypoxia compared with that of normoxic hypoxia. However, the changes of HIF-1 偽 increased rapidly at the early stage of hypoxia, and then decreased rapidly. The expression of HIF-2 偽 and VEGF was the highest in the late stage of hypoxia. After that, the expression of. HIF-1 尾 mRNA and protein remained stable under normoxic or hypoxic conditions. There was no significant change in the expression of HIF-1 尾 mRNA and protein under normoxic or hypoxic conditions. The specific transfection of HIF-1 偽 -HIF-2 偽 and HIF-1 尾 siRNA could effectively silence the expression of their respective genes, and the level of VEGF expression also decreased, and the down-regulation of HIF-2 偽 was the most significant (P 0.05). Conclusion HIF-1 偽 HIF-2 偽 and HIF-1 尾 have different responses to hypoxia stimulation in BGC-823 gastric cancer cells. HIF-1 偽 and HIF-2 偽 may be the main hypoxia inducible factors for gastric cancer to adapt to hypoxia. 2. RNA interference can inhibit the expression of three HIF genes. Compared with HIF-1 偽 and HIF-1 尾 silencing pathway, HIF-2 偽 may be more effective in preventing angiogenesis in gastric cancer than in HIF-1 偽 and HIF-1 尾 silencing pathway.
【學(xué)位授予單位】：新疆醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：R735.2

【參考文獻(xiàn)】

相關(guān)期刊論文 前10條

1 董小鋒;張慶梅;陳元元;李夢陽;周燕秋;青燕;劉天奇;;缺氧誘導(dǎo)因子-2α通過血管生成素-2途徑調(diào)控缺氧誘導(dǎo)血管形成的研究[J];中華消化外科雜志;2016年07期

2 任sズ，

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