LMP-1对人鼻咽癌细胞株CNE1 miRNA表达谱的影响
发布时间：2018-08-05 16:04
【摘要】：目的本研究旨在探讨人鼻咽癌细胞株CNE1、CNE1- LMP1中miRNAs的差异性表达,并预测异常表达的miRNAs可能调控的靶基因。首次探讨LMP-1对鼻咽癌细胞中miRNAs表达谱的影响并对EBV癌基因LMP-1的促癌机理做更深一步的研究,为鼻咽癌的诊断和治疗提供相关的基础数据。 方法培养人鼻咽癌细胞株CNE1和转入了癌基因LMP-1的鼻咽癌细胞株CNE1- LMP1,用免疫组化以及RT-PCR检测CNE1-LMP1中LMP-1的表达,并分别提取细胞总RNA;利用miRNAs芯片技术,检测两种细胞株的miRNAs的表达,对其表达谱进行差异性分析;运用MiRanda、TargetScan、PicTar、DIANA—microT软件预测miRNAs可能调控的靶基因。 结果miRNA在不同的肿瘤细胞中具有特定的表达水平和模式,人们可以通过对人肿瘤组织中的miRNA表达谱与正常的组织表达谱进行对比分析,对不同肿瘤特定的miRNA水平进行鉴定,从而有助于肿瘤的早期诊断和治疗。由于miRNA是基因表达和蛋白质翻译过程中的调节分子,在肿瘤的发生过程中起调控的枢纽作用,因此已有专家预测,把miRNA作为肿瘤生物治疗的靶分子将比编码基因作为靶分子更加有效。LMP-1在EBV潜伏感染的鼻咽癌中有癌基因的特性,在鼻咽癌发生起重要作用。目前在鼻咽癌中EBV编码的LMP-1是否能影响miRNA的表达,从而调节下游靶基因的表达,引起肿瘤细胞生物学行为的改变的研究尚未见报导。我们以miRNAs芯片为平台初步研究了有以及无LMP-1表达的鼻咽癌细胞miRNAs表达谱,在鼻咽癌细胞CNE1和CNE1-LMP1中检出32个存在差异表达的miRNAs。在共同表达的20个分子中,在CNE1-LMP1中表达降低的有8个;hsa-miR-137、hsa-miR-151、hsa-miR-196a、hsa-miR-197、hsa-miR-202、hsa-miR-373、hsa-miR-488和hsa- miR-213;另外12个表达升高,有6个升高达2倍以上:hsa-miR-19b、hsa-miR- 17-3p、hsa-miR-22、hsa-miR-149、hsa-miR-150和hsa-miR-188。对CNE1 -LMP1细胞显著异常表达的miRNAs进行信息学分析,筛选出相应的靶基因。并发现其靶点非常广泛,涉及到癌基因、细胞分化、转录调控、血管生成、信号转导通路等。 结论获得了人鼻咽癌细胞株CNE1、CNE1- LMP1 miRNAs的差异表达谱。其中部分miRNAs可能通过调控其靶基因而参与鼻咽癌的发病机制。
[Abstract]:Objective to investigate the differential expression of miRNAs in human nasopharyngeal carcinoma (NPC) cell line CNE1- LMP1, and to predict the possible target genes regulated by abnormal expression of miRNAs. It is the first time to study the effect of LMP-1 on miRNAs expression profile in nasopharyngeal carcinoma cells and to further study the carcinogenic mechanism of EBV oncogene LMP-1 in order to provide basic data for the diagnosis and treatment of nasopharyngeal carcinoma. Methods Human nasopharyngeal carcinoma cell line CNE1 and nasopharyngeal carcinoma cell line CNE1-LMP1 transfected with oncogene LMP-1 were cultured, the expression of LMP-1 in CNE1-LMP1 was detected by immunohistochemistry and RT-PCR, and the expression of miRNAs was detected by miRNAs microarray technique. The expression profiles were analyzed and the target genes regulated by miRNAs were predicted by MiRanda TargetScang PicTarNA-microT software. Results miRNA had specific expression levels and patterns in different tumor cells. The specific miRNA levels of different tumors could be identified by comparing the expression profiles of miRNA in human tumor tissues with those in normal tissues. It is helpful for early diagnosis and treatment of tumor. Because miRNA is the regulatory molecule in gene expression and protein translation, it plays a pivotal role in the process of tumorigenesis, so it has been predicted by experts. Using miRNA as the target molecule of tumor biotherapy will be more effective than coding gene as target molecule. LMP-1 has the characteristic of oncogene in EBV latent infection nasopharyngeal carcinoma and plays an important role in the occurrence of nasopharyngeal carcinoma. At present, whether the LMP-1 encoded by EBV can affect the expression of miRNA and regulate the expression of downstream target genes in nasopharyngeal carcinoma (NPC) has not been reported. We preliminarily studied the miRNAs expression profiles of NPC cells with and without LMP-1 expression using miRNAs microarray as a platform. 32 differentially expressed miRNAs were detected in CNE1 and CNE1-LMP1 cells. Among the 20 co-expressed molecules, 8 hsa-miR-137nhsa-miR-151nhsa-miR-196aanhsa-miR-197hsa-miR-202 hsa-miR-373hsa-miR-488 and hsa-miR-21313 were down-expressed in CNE1-LMP1, and the other 12 increased in expression, with 6 liters of hsa-miR-19bsa-miR17-3phsa-miR-22hsa-miR-149hsa-miR-150 and hsa-miR-1888. The expression of hsa-miR-137hsa-miR-173psa-miR-173phsa-miR-22hsa-miR-149hsa-miR-150 and hsa-miR-1888. The abnormal expression of miRNAs in CNE1-LMP1 cells was analyzed by informatics, and the target genes were screened out. It is found that the target sites are very extensive, involving oncogene, cell differentiation, transcriptional regulation, angiogenesis, signal transduction pathway and so on. Conclusion the differential expression profile of CNE 1 LMP1 miRNAs in human nasopharyngeal carcinoma cell line CNE 1 was obtained. Some of miRNAs may be involved in the pathogenesis of nasopharyngeal carcinoma by regulating its target gene.
【学位授予单位】：桂林医学院
【学位级别】：硕士
【学位授予年份】：2010
【分类号】：R739.63
本文编号：2166289
[Abstract]:Objective to investigate the differential expression of miRNAs in human nasopharyngeal carcinoma (NPC) cell line CNE1- LMP1, and to predict the possible target genes regulated by abnormal expression of miRNAs. It is the first time to study the effect of LMP-1 on miRNAs expression profile in nasopharyngeal carcinoma cells and to further study the carcinogenic mechanism of EBV oncogene LMP-1 in order to provide basic data for the diagnosis and treatment of nasopharyngeal carcinoma. Methods Human nasopharyngeal carcinoma cell line CNE1 and nasopharyngeal carcinoma cell line CNE1-LMP1 transfected with oncogene LMP-1 were cultured, the expression of LMP-1 in CNE1-LMP1 was detected by immunohistochemistry and RT-PCR, and the expression of miRNAs was detected by miRNAs microarray technique. The expression profiles were analyzed and the target genes regulated by miRNAs were predicted by MiRanda TargetScang PicTarNA-microT software. Results miRNA had specific expression levels and patterns in different tumor cells. The specific miRNA levels of different tumors could be identified by comparing the expression profiles of miRNA in human tumor tissues with those in normal tissues. It is helpful for early diagnosis and treatment of tumor. Because miRNA is the regulatory molecule in gene expression and protein translation, it plays a pivotal role in the process of tumorigenesis, so it has been predicted by experts. Using miRNA as the target molecule of tumor biotherapy will be more effective than coding gene as target molecule. LMP-1 has the characteristic of oncogene in EBV latent infection nasopharyngeal carcinoma and plays an important role in the occurrence of nasopharyngeal carcinoma. At present, whether the LMP-1 encoded by EBV can affect the expression of miRNA and regulate the expression of downstream target genes in nasopharyngeal carcinoma (NPC) has not been reported. We preliminarily studied the miRNAs expression profiles of NPC cells with and without LMP-1 expression using miRNAs microarray as a platform. 32 differentially expressed miRNAs were detected in CNE1 and CNE1-LMP1 cells. Among the 20 co-expressed molecules, 8 hsa-miR-137nhsa-miR-151nhsa-miR-196aanhsa-miR-197hsa-miR-202 hsa-miR-373hsa-miR-488 and hsa-miR-21313 were down-expressed in CNE1-LMP1, and the other 12 increased in expression, with 6 liters of hsa-miR-19bsa-miR17-3phsa-miR-22hsa-miR-149hsa-miR-150 and hsa-miR-1888. The expression of hsa-miR-137hsa-miR-173psa-miR-173phsa-miR-22hsa-miR-149hsa-miR-150 and hsa-miR-1888. The abnormal expression of miRNAs in CNE1-LMP1 cells was analyzed by informatics, and the target genes were screened out. It is found that the target sites are very extensive, involving oncogene, cell differentiation, transcriptional regulation, angiogenesis, signal transduction pathway and so on. Conclusion the differential expression profile of CNE 1 LMP1 miRNAs in human nasopharyngeal carcinoma cell line CNE 1 was obtained. Some of miRNAs may be involved in the pathogenesis of nasopharyngeal carcinoma by regulating its target gene.
【学位授予单位】：桂林医学院
【学位级别】：硕士
【学位授予年份】：2010
【分类号】：R739.63
【引证文献】
相关硕士学位论文 前1条
1 徐涛;microRNA-224通过API-5调节乳腺癌细胞多西紫杉醇耐药性的作用机制的研究[D];南京医科大学;2014年
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