本文選題：胰腺癌 + 長非編碼RNA　； 參考：《華中科技大學(xué)》2016年博士論文

【摘要】：目前研究發(fā)現(xiàn)長非編碼RNA (lncRNAs)參與了包括腫瘤在內(nèi)的多種疾病的發(fā)生發(fā)展過程,但是其在胰腺癌中的具體作用以及潛在的機(jī)制卻仍不明確。本研究發(fā)現(xiàn)一在胰腺癌中扮有重要作用的IncRNA (lncRNA-NUTF2P3-001并且揭示了其潛在的作用機(jī)制。我們首先通過人體胰腺組織lncRNAs基因芯片篩選,發(fā)現(xiàn)了數(shù)個表達(dá)差異顯著的lncRNAs .它們在慢性胰腺炎以及胰腺癌組織中的表達(dá)均較正常胰腺組織明顯升高,其中l(wèi)ncRNA-NUTF2P3-001的表達(dá)水平和臭名昭著的癌基因KRAS mRNA水平呈顯著正相關(guān)。隨后,我們用實時定量逆轉(zhuǎn)錄聚合酶鏈反應(yīng)方法對隨機(jī)選出的30例正常胰腺、30例胰腺癌以及10例慢性胰腺炎組織進(jìn)行了分析以驗證組織芯片的結(jié)果,并對標(biāo)本對應(yīng)的胰腺癌患者進(jìn)行了隨訪。結(jié)果發(fā)現(xiàn)高水平的lncRNA-NUTF2P3-001和較差胰腺癌分級、較晚分期、淋巴侵襲以及遠(yuǎn)處轉(zhuǎn)移等諸多不良預(yù)后息息相關(guān)。然后我們在離體實驗和在體試驗中沉默lncRNA-NUTF2P3-001,結(jié)果發(fā)現(xiàn)下調(diào)lncRNA-NUTF2P3-001可以有效抑制胰腺癌細(xì)胞的活力、生長以及侵襲能力,并伴隨著KRAS以及下游AKT和ERK主要通路活性的改變。之后,雙熒光素酶報告基因試驗結(jié)果證實了lncRNA-NUTF2P3-001以及KRAS mRNA的3’端非轉(zhuǎn)錄區(qū)域(UTR)序列和miR-3923存在競爭性結(jié)合的關(guān)系,并且干擾lncRNA-NUTF2P3-001所引起的腫瘤抑制效果也可以被miR-3923抑制劑消除。因而,實驗結(jié)果明確miR-3923是lncRNA-NUTF2P3-001對KRAS通路調(diào)節(jié)的橋梁。最后我們在離體實驗中發(fā)現(xiàn),低氧或者二氯化鈷處理情況下,HIF-1α可以和lncRNA-NUTF2P3-001啟動子區(qū)域中的低氧反映元件(HREs)結(jié)合進(jìn)而上調(diào)lncRNA.NUTF2P3.001在胰腺癌細(xì)胞中的表達(dá)。因此,該研究提出一全新的低氧刺激一IncRNAs— miRNAs—KRAS調(diào)控通路并且揭示了lncRNA.NUTF2P3.001以及miR-3923對胰腺癌潛在的治療效果,為胰腺癌治療提供了新的線索。
[Abstract]:Current studies have found that long non coded RNA (lncRNAs) is involved in the development of a variety of diseases, including tumors, but its specific role in pancreatic cancer and the underlying mechanisms are still unclear. This study found a IncRNA (lncRNA-NUTF2P3-001, which plays an important role in pancreatic cancer (lncRNA-NUTF2P3-001) and reveals its potential role. We first screened the human pancreatic tissue lncRNAs gene chip and found several lncRNAs with significant differences in expression. Their expression in chronic pancreatitis and pancreatic cancer tissues was significantly higher than that in normal pancreatic tissue. The expression level of lncRNA-NUTF2P3-001 and the level of the infamous oncogene KRAS mRNA were significantly positive. Subsequently, we used real time quantitative reverse transcriptase polymerase chain reaction to analyze 30 randomly selected normal pancreas, 30 cases of pancreatic cancer and 10 cases of chronic pancreatitis to verify the results of the tissue microarray and follow up the corresponding pancreatic cancer patients. The results were found to be high level of lncRNA-NUTF2P3-001 and poor. Pancreatic cancer classification, late stages, lymphatic invasion and distant metastasis are closely related. Then we silenced lncRNA-NUTF2P3-001 in vitro and in vivo tests. The results showed that down-regulation of lncRNA-NUTF2P3-001 could effectively inhibit the vitality, growth and invasion of pancreatic cancer cells, accompanied by KRAS and downstream AKT. After the changes in the activity of the main pathway of ERK, the results of the double luciferase reporter gene test confirmed the competitive binding of the 3 'terminal non transcriptional region (UTR) sequence of the lncRNA-NUTF2P3-001 and the KRAS mRNA, and the inhibition effect of the swelling of the tumor caused by the interference of lncRNA-NUTF2P3-001 can also be eliminated by the miR-3923 inhibitors. The results showed that miR-3923 was the bridge of the lncRNA-NUTF2P3-001 regulation of the KRAS pathway. Finally, in the isolated experiment, we found that HIF-1 alpha could be combined with the low oxygen reflection element (HREs) in the lncRNA-NUTF2P3-001 promoter region to increase the table of the lncRNA.NUTF2P3.001 in the pancreatic cancer cells in the case of hypoxia or two cobalt chloride treatment. So, the study proposed a new hypoxic stimulation of a IncRNAs - miRNAs - KRAS regulatory pathway and revealed the potential therapeutic effects of lncRNA.NUTF2P3.001 and miR-3923 on pancreatic cancer, providing new clues for the treatment of pancreatic cancer.

【學(xué)位授予單位】：華中科技大學(xué)
【學(xué)位級別】：博士
【學(xué)位授予年份】：2016
【分類號】：R735.9

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1 李想;低氧誘導(dǎo)的長非編碼RNA-NUTF2P3-001通過抑制miR-3923對KRAS的降解促進(jìn)胰腺癌發(fā)展[D];華中科技大學(xué);2016年

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