LMP-1對(duì)人鼻咽癌細(xì)胞株CNE1 miRNA表達(dá)譜的影響
發(fā)布時(shí)間：2018-08-05 16:04
【摘要】：目的本研究旨在探討人鼻咽癌細(xì)胞株CNE1、CNE1- LMP1中miRNAs的差異性表達(dá),并預(yù)測(cè)異常表達(dá)的miRNAs可能調(diào)控的靶基因。首次探討LMP-1對(duì)鼻咽癌細(xì)胞中miRNAs表達(dá)譜的影響并對(duì)EBV癌基因LMP-1的促癌機(jī)理做更深一步的研究,為鼻咽癌的診斷和治療提供相關(guān)的基礎(chǔ)數(shù)據(jù)。 方法培養(yǎng)人鼻咽癌細(xì)胞株CNE1和轉(zhuǎn)入了癌基因LMP-1的鼻咽癌細(xì)胞株CNE1- LMP1,用免疫組化以及RT-PCR檢測(cè)CNE1-LMP1中LMP-1的表達(dá),并分別提取細(xì)胞總RNA;利用miRNAs芯片技術(shù),檢測(cè)兩種細(xì)胞株的miRNAs的表達(dá),對(duì)其表達(dá)譜進(jìn)行差異性分析;運(yùn)用MiRanda、TargetScan、PicTar、DIANA—microT軟件預(yù)測(cè)miRNAs可能調(diào)控的靶基因。 結(jié)果miRNA在不同的腫瘤細(xì)胞中具有特定的表達(dá)水平和模式,人們可以通過對(duì)人腫瘤組織中的miRNA表達(dá)譜與正常的組織表達(dá)譜進(jìn)行對(duì)比分析,對(duì)不同腫瘤特定的miRNA水平進(jìn)行鑒定,從而有助于腫瘤的早期診斷和治療。由于miRNA是基因表達(dá)和蛋白質(zhì)翻譯過程中的調(diào)節(jié)分子,在腫瘤的發(fā)生過程中起調(diào)控的樞紐作用,因此已有專家預(yù)測(cè),把miRNA作為腫瘤生物治療的靶分子將比編碼基因作為靶分子更加有效。LMP-1在EBV潛伏感染的鼻咽癌中有癌基因的特性,在鼻咽癌發(fā)生起重要作用。目前在鼻咽癌中EBV編碼的LMP-1是否能影響miRNA的表達(dá),從而調(diào)節(jié)下游靶基因的表達(dá),引起腫瘤細(xì)胞生物學(xué)行為的改變的研究尚未見報(bào)導(dǎo)。我們以miRNAs芯片為平臺(tái)初步研究了有以及無LMP-1表達(dá)的鼻咽癌細(xì)胞miRNAs表達(dá)譜,在鼻咽癌細(xì)胞CNE1和CNE1-LMP1中檢出32個(gè)存在差異表達(dá)的miRNAs。在共同表達(dá)的20個(gè)分子中,在CNE1-LMP1中表達(dá)降低的有8個(gè);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個(gè)表達(dá)升高,有6個(gè)升高達(dá)2倍以上:hsa-miR-19b、hsa-miR- 17-3p、hsa-miR-22、hsa-miR-149、hsa-miR-150和hsa-miR-188。對(duì)CNE1 -LMP1細(xì)胞顯著異常表達(dá)的miRNAs進(jìn)行信息學(xué)分析,篩選出相應(yīng)的靶基因。并發(fā)現(xiàn)其靶點(diǎn)非常廣泛,涉及到癌基因、細(xì)胞分化、轉(zhuǎn)錄調(diào)控、血管生成、信號(hào)轉(zhuǎn)導(dǎo)通路等。 結(jié)論獲得了人鼻咽癌細(xì)胞株CNE1、CNE1- LMP1 miRNAs的差異表達(dá)譜。其中部分miRNAs可能通過調(diào)控其靶基因而參與鼻咽癌的發(fā)病機(jī)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.
【學(xué)位授予單位】：桂林醫(yī)學(xué)院
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2010
【分類號(hào)】：R739.63
本文編號(hào)：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.
【學(xué)位授予單位】：桂林醫(yī)學(xué)院
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2010
【分類號(hào)】：R739.63
【引證文獻(xiàn)】
相關(guān)碩士學(xué)位論文 前1條
1 徐濤;microRNA-224通過API-5調(diào)節(jié)乳腺癌細(xì)胞多西紫杉醇耐藥性的作用機(jī)制的研究[D];南京醫(yī)科大學(xué);2014年
，本文編號(hào)：2166289
本文鏈接：http://sikaile.net/yixuelunwen/yank/2166289.html
最近更新
教材專著
