基因芯片技術(shù)用于鼻咽癌及其放療敏感性的研究
發(fā)布時(shí)間：2018-07-31 19:41
【摘要】：鼻咽癌是一種上皮源性惡性腫瘤,絕大多數(shù)屬于低分化鱗狀細(xì)胞癌,惡性程度高,早期易發(fā)生淋巴結(jié)和全身遠(yuǎn)處轉(zhuǎn)移,具有相當(dāng)高的死亡率。在一些地區(qū),特別是中國南部地區(qū),鼻咽癌的發(fā)生非常普遍,其發(fā)生率是世界其他地區(qū)的10-30倍。已知的鼻咽癌的病因主要包括基因的易感性,環(huán)境中的致癌的化學(xué)物質(zhì),以及EB病毒的感染。放療是鼻咽癌治療的主要方法,而放療后5年的生存率只有50-60%。在大多數(shù)病例中放療抑制性成為阻礙治療的一個(gè)重大障礙。所以找出放療敏感型和抑制型之間的差異,提前預(yù)測(cè)放療抑制型的病人,做出新的治療方案能為鼻咽癌的治療提供必要的幫助。 我們利用能夠檢測(cè)14112個(gè)人類基因的cDNA芯片,對(duì)20例鼻咽癌病人組織樣本與15例鼻咽慢性炎癥病人組織樣本進(jìn)行表達(dá)譜分析比較。我們用ScanArray4000對(duì)芯片進(jìn)行掃描,GenePix Pro3.0軟件被用于芯片型號(hào)強(qiáng)度的分析和篩選。在完成芯片數(shù)據(jù)的歸一化及篩選修補(bǔ)處理之后,利用SAM軟件及Cluster軟件進(jìn)行差異基因的分析及聚類分析。 表達(dá)譜芯片的分析結(jié)果顯示,9個(gè)基因(q0.01)至少在17例(85%)鼻咽癌組織中有2倍差異表達(dá),其中8個(gè)下調(diào)基因(TAOK3, SLC16A2, PRB4, AMY2B, B3GALT4, MSMB, RPS27, CR2),1個(gè)上調(diào)基因(MXLIP)。為了進(jìn)一步研究,我們選用CR2和MXLIP在另外50例低分化鼻咽癌組織中進(jìn)行real-time RT-PCR驗(yàn)證。通過另外3例鼻咽慢性炎癥組織的表達(dá)譜比較,real-time RT-PCR結(jié)果與芯片實(shí)驗(yàn)結(jié)果相同,CR2在41例(82%)鼻咽癌組織中下調(diào),MXLIP在42例(84%)鼻咽癌組織中上調(diào)。 隨后,我們將這20例鼻咽癌病例對(duì)放療敏感性進(jìn)行分類,有8例鼻咽癌敏感型的病人組織樣本與12例鼻咽癌放療抑制型的病人組織樣本。通過對(duì)這兩組病人的表達(dá)譜數(shù)據(jù)進(jìn)行比較,我們篩選出了111條在鼻咽癌放療抑制型組織和敏感型組織中差異表達(dá)的基因,其中包括108條在抑制型中上調(diào)基因(ZNF608,PIZEO2, CSF1R等)和3條在抑制型中下調(diào)基因(ATP2C1, MUDENG, OLA1)。為了驗(yàn)證芯片結(jié)果的可靠性,我們應(yīng)用定量RT-PCR的方法,在另外17例鼻咽癌病人樣本中檢測(cè)了差異表達(dá)基因ZNF608以及CSF1R的表達(dá)情況,RT-PCR結(jié)果與芯片結(jié)果吻合。 我們將這些差異表達(dá)基因輸入GenMAPP軟件進(jìn)行生物學(xué)通路分析。在通路分析中,我們發(fā)現(xiàn)了9類26條生物學(xué)通路與鼻咽癌的放療抑制性有關(guān)。大部分靶基因富集與細(xì)胞離子平衡,細(xì)胞因子及免疫反應(yīng),體液免疫,細(xì)胞增殖,受體蛋白信號(hào)途徑等通路。這里,我們認(rèn)為鼻咽癌的放療抑制性可能主要是由于細(xì)胞內(nèi)鈣離子的變化引起的。它能在放療時(shí)抑制細(xì)胞凋亡,促進(jìn)DNA修復(fù)達(dá)到拯救癌細(xì)胞的作用。另外,各種細(xì)胞內(nèi)源性或者外源性因素引起的細(xì)胞增殖起到在治療時(shí)維持腫瘤的大小的作用。
[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.
【學(xué)位授予單位】：復(fù)旦大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2011
【分類號(hào)】：R739.63
本文編號(hào)：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.
【學(xué)位授予單位】：復(fù)旦大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2011
【分類號(hào)】：R739.63
【參考文獻(xiàn)】
相關(guān)期刊論文 前2條
1 賀玉香;仲萍萍;嚴(yán)珊珊;劉莉;史弘流;曾木圣;夏云飛;;DNA-PK的活性與鼻咽癌細(xì)胞株CNE1/CNE2放射敏感性的關(guān)系(英文)[J];生理學(xué)報(bào);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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