低氧诱导的长非编码RNA-NUTF2P3-001通过抑制miR-3923对KRAS的降解促进胰腺癌发展

发布时间：2018-04-21 06:40

本文选题：胰腺癌 + 长非编码RNA　；参考：《华中科技大学》2016年博士论文

【摘要】：目前研究发现长非编码RNA (lncRNAs)参与了包括肿瘤在内的多种疾病的发生发展过程,但是其在胰腺癌中的具体作用以及潜在的机制却仍不明确。本研究发现一在胰腺癌中扮有重要作用的IncRNA (lncRNA-NUTF2P3-001并且揭示了其潜在的作用机制。我们首先通过人体胰腺组织lncRNAs基因芯片筛选,发现了数个表达差异显著的lncRNAs .它们在慢性胰腺炎以及胰腺癌组织中的表达均较正常胰腺组织明显升高,其中lncRNA-NUTF2P3-001的表达水平和臭名昭著的癌基因KRAS mRNA水平呈显著正相关。随后,我们用实时定量逆转录聚合酶链反应方法对随机选出的30例正常胰腺、30例胰腺癌以及10例慢性胰腺炎组织进行了分析以验证组织芯片的结果,并对标本对应的胰腺癌患者进行了随访。结果发现高水平的lncRNA-NUTF2P3-001和较差胰腺癌分级、较晚分期、淋巴侵袭以及远处转移等诸多不良预后息息相关。然后我们在离体实验和在体试验中沉默lncRNA-NUTF2P3-001,结果发现下调lncRNA-NUTF2P3-001可以有效抑制胰腺癌细胞的活力、生长以及侵袭能力,并伴随着KRAS以及下游AKT和ERK主要通路活性的改变。之后,双荧光素酶报告基因试验结果证实了lncRNA-NUTF2P3-001以及KRAS mRNA的3’端非转录区域(UTR)序列和miR-3923存在竞争性结合的关系,并且干扰lncRNA-NUTF2P3-001所引起的肿瘤抑制效果也可以被miR-3923抑制剂消除。因而,实验结果明确miR-3923是lncRNA-NUTF2P3-001对KRAS通路调节的桥梁。最后我们在离体实验中发现,低氧或者二氯化钴处理情况下,HIF-1α可以和lncRNA-NUTF2P3-001启动子区域中的低氧反映元件(HREs)结合进而上调lncRNA.NUTF2P3.001在胰腺癌细胞中的表达。因此,该研究提出一全新的低氧刺激一IncRNAs— miRNAs—KRAS调控通路并且揭示了lncRNA.NUTF2P3.001以及miR-3923对胰腺癌潜在的治疗效果,为胰腺癌治疗提供了新的线索。
[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.

【学位授予单位】：华中科技大学
【学位级别】：博士
【学位授予年份】：2016
【分类号】：R735.9

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