基因芯片技术用于鼻咽癌及其放疗敏感性的研究
发布时间：2018-07-31 19:41
【摘要】：鼻咽癌是一种上皮源性恶性肿瘤,绝大多数属于低分化鳞状细胞癌,恶性程度高,早期易发生淋巴结和全身远处转移,具有相当高的死亡率。在一些地区,特别是中国南部地区,鼻咽癌的发生非常普遍,其发生率是世界其他地区的10-30倍。已知的鼻咽癌的病因主要包括基因的易感性,环境中的致癌的化学物质,以及EB病毒的感染。放疗是鼻咽癌治疗的主要方法,而放疗后5年的生存率只有50-60%。在大多数病例中放疗抑制性成为阻碍治疗的一个重大障碍。所以找出放疗敏感型和抑制型之间的差异,提前预测放疗抑制型的病人,做出新的治疗方案能为鼻咽癌的治疗提供必要的帮助。 我们利用能够检测14112个人类基因的cDNA芯片,对20例鼻咽癌病人组织样本与15例鼻咽慢性炎症病人组织样本进行表达谱分析比较。我们用ScanArray4000对芯片进行扫描,GenePix Pro3.0软件被用于芯片型号强度的分析和筛选。在完成芯片数据的归一化及筛选修补处理之后,利用SAM软件及Cluster软件进行差异基因的分析及聚类分析。 表达谱芯片的分析结果显示,9个基因(q0.01)至少在17例(85%)鼻咽癌组织中有2倍差异表达,其中8个下调基因(TAOK3, SLC16A2, PRB4, AMY2B, B3GALT4, MSMB, RPS27, CR2),1个上调基因(MXLIP)。为了进一步研究,我们选用CR2和MXLIP在另外50例低分化鼻咽癌组织中进行real-time RT-PCR验证。通过另外3例鼻咽慢性炎症组织的表达谱比较,real-time RT-PCR结果与芯片实验结果相同,CR2在41例(82%)鼻咽癌组织中下调,MXLIP在42例(84%)鼻咽癌组织中上调。 随后,我们将这20例鼻咽癌病例对放疗敏感性进行分类,有8例鼻咽癌敏感型的病人组织样本与12例鼻咽癌放疗抑制型的病人组织样本。通过对这两组病人的表达谱数据进行比较,我们筛选出了111条在鼻咽癌放疗抑制型组织和敏感型组织中差异表达的基因,其中包括108条在抑制型中上调基因(ZNF608,PIZEO2, CSF1R等)和3条在抑制型中下调基因(ATP2C1, MUDENG, OLA1)。为了验证芯片结果的可靠性,我们应用定量RT-PCR的方法,在另外17例鼻咽癌病人样本中检测了差异表达基因ZNF608以及CSF1R的表达情况,RT-PCR结果与芯片结果吻合。 我们将这些差异表达基因输入GenMAPP软件进行生物学通路分析。在通路分析中,我们发现了9类26条生物学通路与鼻咽癌的放疗抑制性有关。大部分靶基因富集与细胞离子平衡,细胞因子及免疫反应,体液免疫,细胞增殖,受体蛋白信号途径等通路。这里,我们认为鼻咽癌的放疗抑制性可能主要是由于细胞内钙离子的变化引起的。它能在放疗时抑制细胞凋亡,促进DNA修复达到拯救癌细胞的作用。另外,各种细胞内源性或者外源性因素引起的细胞增殖起到在治疗时维持肿瘤的大小的作用。
[Abstract]:Nasopharyngeal carcinoma (NPC) is an epithelial-derived malignant tumor, most of which belong to poorly differentiated squamous cell carcinoma. Nasopharyngeal carcinoma is common in some regions, especially in southern China, and is 10-30 times higher than in other parts of the world. Known causes of nasopharyngeal carcinoma include genetic susceptibility, carcinogenic chemicals in the environment, and Epstein-Barr virus infection. Radiotherapy is the main treatment for nasopharyngeal carcinoma, but the 5-year survival rate after radiotherapy is only 50-60. In most cases, the inhibition of radiotherapy is a major obstacle to treatment. Therefore, to find out the difference between the sensitive and inhibitory types of radiotherapy, to predict the patients with the type of radiotherapy in advance, and to make a new treatment plan can provide necessary help for the treatment of nasopharyngeal carcinoma. Using cDNA microarray which can detect 14112 human genes, we analyzed and compared the expression profiles of 20 nasopharyngeal carcinoma tissue samples and 15 nasopharyngeal chronic inflammation tissue samples. We use ScanArray4000 to scan the chip. GenePix Pro3.0 software is used to analyze and screen the model strength of the chip. After the normalization of chip data and the screening and mending, the differential gene analysis and cluster analysis were carried out by using SAM and Cluster software. The results of microarray analysis showed that 9 genes (q0.01) were twice differentially expressed in at least 17 cases (85%) of nasopharyngeal carcinoma tissues, 8 down-regulated genes (TAOK3, SLC16A2, PRB4, AMY2B, B3GALT4, MSMBR, RPS27, CR2), and 1 up-regulated gene (MXLIP). For further study, we selected CR2 and MXLIP to perform real-time RT-PCR validation in another 50 cases of poorly differentiated nasopharyngeal carcinoma. The results of real-time RT-PCR were the same as those of microarray in 41 cases (82%) of nasopharyngeal carcinoma. The down-regulation of MXLIP was up-regulated in 42 cases (84%) of nasopharyngeal carcinoma. Subsequently, we classified the radiosensitivity of 20 patients with nasopharyngeal carcinoma (NPC), including 8 patients with nasopharyngeal carcinoma (NPC) sensitive type and 12 patients with nasopharyngeal carcinoma (NPC). By comparing the expression profile data of the two groups of patients, we screened 111 differentially expressed genes in tumor suppressive and sensitive nasopharyngeal carcinoma tissues. There were 108 up-regulated genes (ZNF608, PIZEO _ 2, CSF1R, etc.) and 3 down-regulated genes (ATP2C1, MUDENG, OLA1). In order to verify the reliability of the microarray results, we used quantitative RT-PCR method to detect the differential expression gene ZNF608 and the expression of CSF1R in 17 patients with nasopharyngeal carcinoma. The results of RT-PCR were in agreement with the results of the microarray. We input these differentially expressed genes into GenMAPP software for biological pathway analysis. In the pathway analysis, we found that 9 types of 26 biological pathways were associated with radiotherapy inhibition in nasopharyngeal carcinoma. Most of the target gene enrichment is associated with cellular ion balance, cytokine and immunoreaction, humoral immunity, cell proliferation, receptor protein signaling pathway, and so on. Here, we suggest that the inhibition of radiotherapy in nasopharyngeal carcinoma may be mainly due to changes in intracellular calcium. It can inhibit cell apoptosis during radiotherapy and promote DNA repair to save cancer cells. In addition, cell proliferation caused by various cellular endogenous or exogenous factors plays a role in maintaining tumor size during treatment.
【学位授予单位】：复旦大学
【学位级别】：硕士
【学位授予年份】：2011
【分类号】：R739.63
本文编号：2156630
[Abstract]:Nasopharyngeal carcinoma (NPC) is an epithelial-derived malignant tumor, most of which belong to poorly differentiated squamous cell carcinoma. Nasopharyngeal carcinoma is common in some regions, especially in southern China, and is 10-30 times higher than in other parts of the world. Known causes of nasopharyngeal carcinoma include genetic susceptibility, carcinogenic chemicals in the environment, and Epstein-Barr virus infection. Radiotherapy is the main treatment for nasopharyngeal carcinoma, but the 5-year survival rate after radiotherapy is only 50-60. In most cases, the inhibition of radiotherapy is a major obstacle to treatment. Therefore, to find out the difference between the sensitive and inhibitory types of radiotherapy, to predict the patients with the type of radiotherapy in advance, and to make a new treatment plan can provide necessary help for the treatment of nasopharyngeal carcinoma. Using cDNA microarray which can detect 14112 human genes, we analyzed and compared the expression profiles of 20 nasopharyngeal carcinoma tissue samples and 15 nasopharyngeal chronic inflammation tissue samples. We use ScanArray4000 to scan the chip. GenePix Pro3.0 software is used to analyze and screen the model strength of the chip. After the normalization of chip data and the screening and mending, the differential gene analysis and cluster analysis were carried out by using SAM and Cluster software. The results of microarray analysis showed that 9 genes (q0.01) were twice differentially expressed in at least 17 cases (85%) of nasopharyngeal carcinoma tissues, 8 down-regulated genes (TAOK3, SLC16A2, PRB4, AMY2B, B3GALT4, MSMBR, RPS27, CR2), and 1 up-regulated gene (MXLIP). For further study, we selected CR2 and MXLIP to perform real-time RT-PCR validation in another 50 cases of poorly differentiated nasopharyngeal carcinoma. The results of real-time RT-PCR were the same as those of microarray in 41 cases (82%) of nasopharyngeal carcinoma. The down-regulation of MXLIP was up-regulated in 42 cases (84%) of nasopharyngeal carcinoma. Subsequently, we classified the radiosensitivity of 20 patients with nasopharyngeal carcinoma (NPC), including 8 patients with nasopharyngeal carcinoma (NPC) sensitive type and 12 patients with nasopharyngeal carcinoma (NPC). By comparing the expression profile data of the two groups of patients, we screened 111 differentially expressed genes in tumor suppressive and sensitive nasopharyngeal carcinoma tissues. There were 108 up-regulated genes (ZNF608, PIZEO _ 2, CSF1R, etc.) and 3 down-regulated genes (ATP2C1, MUDENG, OLA1). In order to verify the reliability of the microarray results, we used quantitative RT-PCR method to detect the differential expression gene ZNF608 and the expression of CSF1R in 17 patients with nasopharyngeal carcinoma. The results of RT-PCR were in agreement with the results of the microarray. We input these differentially expressed genes into GenMAPP software for biological pathway analysis. In the pathway analysis, we found that 9 types of 26 biological pathways were associated with radiotherapy inhibition in nasopharyngeal carcinoma. Most of the target gene enrichment is associated with cellular ion balance, cytokine and immunoreaction, humoral immunity, cell proliferation, receptor protein signaling pathway, and so on. Here, we suggest that the inhibition of radiotherapy in nasopharyngeal carcinoma may be mainly due to changes in intracellular calcium. It can inhibit cell apoptosis during radiotherapy and promote DNA repair to save cancer cells. In addition, cell proliferation caused by various cellular endogenous or exogenous factors plays a role in maintaining tumor size during treatment.
【学位授予单位】：复旦大学
【学位级别】：硕士
【学位授予年份】：2011
【分类号】：R739.63
【参考文献】
相关期刊论文 前2条
1 贺玉香;仲萍萍;严珊珊;刘莉;史弘流;曾木圣;夏云飞;;DNA-PK的活性与鼻咽癌细胞株CNE1/CNE2放射敏感性的关系(英文)[J];生理学报;2007年04期
2 James T Taylor;Jonathan E Pottle;Kevin Lee;Alun R Wang;Stephenie G Yi;Jennifer A S Scruggs;Suresh S Sikka;;Calcium signaling and T-type calcium channels in cancer cell cycling[J];World Journal of Gastroenterology;2008年32期
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