基于全转录组测序技术对人肺腺癌A549细胞顺铂获得性耐药机制的研究
发布时间：2018-08-05 09:23
【摘要】：治疗过程中产生的顺铂耐药现象是影响非小细胞肺癌患者临床化疗的重要因素之一,但其潜在的分子机制尚未完全阐明。本研究旨在发现与顺铂获得性耐药相关的关键基因及信号通路。基于全转录组测序技术,我们对非小细胞肺癌A549细胞及相应的顺铂耐药细胞A549/DDP的差异表达基因谱进行了系统分析。转录组测序共获得1214个差异表达的基因,其中656个基因在A549/DDP细胞中表达上调,588个基因表达下调。基于KEGG数据库的数据挖掘表明,富集最多差异表达基因的信号通路为PI3K/AKT,MAPK,Focal adhesion 与 Actin cytoskeleton regulation,并且这些通路的关键节点基因均存在显著差异表达的情况,表明这些信号通路在调节非小细胞肺癌顺铂耐药方面发挥着关键的作用。除以上几大信号通路外,本研究还发现醛酮还原酶家族l(Aldo-Keto Reductases family 1,AKR1)的三个成员AKR1C1,AKR1C3及AKR1B1同时在A549/DDP细胞中显著上调,且AKR1C1基因是差异表达最为显著的基因之一。功能性实验证实,AKR1C1或AKR1B1酶活抑制剂与顺铂联合使用可显著提高A549/DDP细胞的顺铂敏感性;此外,在A549/DDP细胞中同时敲减AKR1C1和AKR1B1不仅增强了顺铂的细胞毒性,且同时显著增强了顺铂诱导的细胞凋亡作用。以上结果表明,AKR1家族中的AKR1C1和AKR1B1基因在调节A549细胞顺铂耐药中起到了关键的作用。本研究为非小细胞肺癌临床治疗提供了新的分子靶标,在临床样本中验证本研究发现的分子靶标将是我们下一步工作的重点。
[Abstract]:Cisplatin resistance is one of the important factors affecting the clinical chemotherapy of patients with non-small cell lung cancer, but its potential molecular mechanism has not been fully elucidated. This study aims to identify key genes and signaling pathways associated with cisplatin acquired resistance. Based on the whole transcriptome sequencing technique, we systematically analyzed the differentially expressed gene profiles of A549/DDP in A549 cells and cisplatin resistant cells of non-small cell lung cancer (NSCLC). A total of 1214 differentially expressed genes were obtained by transcriptome sequencing, of which 656 genes were up-regulated and down-regulated in A549/DDP cells. Data mining based on KEGG database showed that the signal pathways that enriched the most differentially expressed genes were PI3K / AKTK-MAPKFocal adhesion and Actin cytoskeleton regulation, and the key node genes of these pathways were significantly differentially expressed. These signaling pathways play a key role in regulating cisplatin resistance in non-small cell lung cancer. In addition to these major signal pathways, we also found that AKR1C1AK-R1C3 and AKR1B1, three members of the aldehyde-ketone reductase family (Aldo-Keto Reductases family 1), were up-regulated in A549/DDP cells at the same time, and AKR1C1 gene was one of the most significant differentially expressed genes. Functional experiments showed that the combination of AKR1C1 or AKR1B1 enzyme inhibitor with cisplatin could significantly increase the cisplatin sensitivity of A549/DDP cells, in addition, knockout of AKR1C1 and AKR1B1 in A549/DDP cells not only enhanced the cytotoxicity of cisplatin, but also increased the cytotoxicity of cisplatin. At the same time, the apoptosis induced by cisplatin was significantly enhanced. These results suggest that the AKR1C1 and AKR1B1 genes in the AKR1 family play a key role in regulating cisplatin resistance in A549 cells. This study provides a new molecular target for the clinical treatment of non-small cell lung cancer (NSCLC).
【学位授予单位】：西北大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R734.2
本文编号：2165352
[Abstract]:Cisplatin resistance is one of the important factors affecting the clinical chemotherapy of patients with non-small cell lung cancer, but its potential molecular mechanism has not been fully elucidated. This study aims to identify key genes and signaling pathways associated with cisplatin acquired resistance. Based on the whole transcriptome sequencing technique, we systematically analyzed the differentially expressed gene profiles of A549/DDP in A549 cells and cisplatin resistant cells of non-small cell lung cancer (NSCLC). A total of 1214 differentially expressed genes were obtained by transcriptome sequencing, of which 656 genes were up-regulated and down-regulated in A549/DDP cells. Data mining based on KEGG database showed that the signal pathways that enriched the most differentially expressed genes were PI3K / AKTK-MAPKFocal adhesion and Actin cytoskeleton regulation, and the key node genes of these pathways were significantly differentially expressed. These signaling pathways play a key role in regulating cisplatin resistance in non-small cell lung cancer. In addition to these major signal pathways, we also found that AKR1C1AK-R1C3 and AKR1B1, three members of the aldehyde-ketone reductase family (Aldo-Keto Reductases family 1), were up-regulated in A549/DDP cells at the same time, and AKR1C1 gene was one of the most significant differentially expressed genes. Functional experiments showed that the combination of AKR1C1 or AKR1B1 enzyme inhibitor with cisplatin could significantly increase the cisplatin sensitivity of A549/DDP cells, in addition, knockout of AKR1C1 and AKR1B1 in A549/DDP cells not only enhanced the cytotoxicity of cisplatin, but also increased the cytotoxicity of cisplatin. At the same time, the apoptosis induced by cisplatin was significantly enhanced. These results suggest that the AKR1C1 and AKR1B1 genes in the AKR1 family play a key role in regulating cisplatin resistance in A549 cells. This study provides a new molecular target for the clinical treatment of non-small cell lung cancer (NSCLC).
【学位授予单位】：西北大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R734.2
【参考文献】
相关期刊论文 前1条
1 Valentina Grossi;Alessia Peserico;Tugsan Tezil;Cristiano Simone;;p38α MAPK pathway:A key factor in colorectal cancer therapy and chemoresistance[J];World Journal of Gastroenterology;2014年29期
，本文编号：2165352
本文链接：https://www.wllwen.com/yixuelunwen/zlx/2165352.html
