microRNA在非小细胞肺癌发生、发展进程中调控机制的研究

发布时间：2018-07-05 06:57

本文选题：非小细胞肺癌 + miR-199a-5p　；参考：《上海大学》2016年博士论文

【摘要】：非小细胞肺癌占肺癌总数的80-85%,并伴有极高的死亡率,早期诊断可以改善这一现状。在多种NSCLC亚型中,经常检测出KRAS基因被激活和抑癌基因LKB1/P53失活。KRAS基因激活同时伴有LKB1/P53缺失的NSCLC恶性程度显著升高。但是,LKB1/P53缺失后的NSCLC特征性miRNA表达谱,以及这些miRNAs在肿瘤中的作用机制尚未系统研究。本研究中,我们通过solexa测序分析了正常小鼠(C57BL/6)、良性肿瘤小鼠模型(KRASG12D)和恶性肿瘤模型(KRASG12D/P53-/-和KRASG12D/LKB1-/-)的肺组织中差异表达的miRNAs。利用生物信息学的方法分析了30个持续性表达上调和24个持续性表达下调miRNAs的靶基因及靶基因富集的信号通路,并进一步分析了这些靶蛋白之间的蛋白互作关系。随后在54种与对照小鼠肺组织相比表达量下调的miRNAs中,选取miR-10a和miR-199a-5p在NSCLC细胞系中作进一步验证。研究发现,miR-199a-5p和miR-10a的表达量在小鼠肺癌组织模型和人临床组织样本中显著降低。研究证明,miR-199a-5p在NSCLC细胞中通过靶向MAP3K11抑制细胞的增殖和周期。miR-10a则通过抑制MAP3K7和WNK3抑制非小细胞肺癌的增殖,促进细胞凋亡。研究一并证实了P53可以在肺腺癌细胞系SPCA-1中调控miR-10a的转录。P53作为重要的转录因子在肿瘤发生进程中扮演重要角色。研究发现,miR-150可以通过靶向P53 mRNA的3'-UTR调控P53的表达,进而影响细胞周期。P53缺失的H1299细胞中过表达P53后,检测发现miR-34a,miR-184,miR-181a和miR-148的表达量显著升高。综上研究结果表明,miR-150,P53以及相关miNRAs在NSCLC中组成网络,调控NSCLC的发生和发展。
[Abstract]:Non-small cell lung cancer (NSCLC) accounts for 80-85% of lung cancer and has a high mortality rate. Early diagnosis can improve the status quo. In many NSCLC subtypes, KRAS gene activation and tumor suppressor gene LKB1 / p53 inactivation. KRAS gene activation accompanied by LKB1 / p53 deletion were frequently detected in NSCLC, and the malignant degree of NSCLC with LKB1 / p53 deletion was significantly increased. However, the characteristic miRNA expression profiles of NSCLC without LKB1 / p53 deletion and the mechanism of these miRNAs in tumor have not been systematically studied. In this study, we sequenced miRNAs differentially expressed in lung tissues of normal mice (C57BL / 6), benign tumor mice (KRASG12D) and malignant tumor models (KRASG12D / P53-r- and KRASG12D / LKB1-r-) by solexa sequencing. A bioinformatics method was used to analyze 30 up-regulated and 24 persistent down-regulated target genes of miRNAs and the signal pathways enriched by target genes, and the protein interaction between these proteins was further analyzed. Then miR-10a and miR-199a-5p were selected for further verification in NSCLC cell line. It was found that the expression of miR-199a-5p and miR-10a decreased significantly in mouse lung cancer tissue model and human clinical tissue samples. The results showed that miR-199a-5p inhibited the proliferation of NSCLC cells by targeting MAP3K11 and the cell cycle. MiR-10a inhibited the proliferation of non-small cell lung cancer by inhibiting MAP3K7 and WNK3 and promoted the apoptosis of NSCLC cells. It has been confirmed that p53 can regulate miR-10a transcription in lung adenocarcinoma cell line SPCA-1. As an important transcription factor, p53 plays an important role in tumorigenesis. It was found that miR-150 could regulate the expression of p53 by targeting p53 mRNA, and then affect the overexpression of p53 in H1299 cells with cell cycle. It was found that the expression of miR-184miR-181a and miR-148 increased significantly in H1299 cells. The results show that the network of NSCLC is composed of P53 and related miNRAs, which regulate the occurrence and development of NSCLC.
【学位授予单位】：上海大学
【学位级别】：博士
【学位授予年份】：2016
【分类号】：R734.2

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