本文選題：NFBD1　切入點：鼻咽癌　出處：《重慶醫(yī)科大學》2012年碩士論文

【摘要】：目的：了解NFBD1在鼻咽癌組織中mRNA及蛋白質(zhì)的表達水平，及蛋白在細胞內(nèi)的分布，并研究siRNA介導的鼻咽癌細胞株中NFBD1的表達抑制后，對鼻咽癌細胞株化療敏感性的影響。 方法：收集臨床已確診的鼻咽癌組織與正常鼻咽部組織，提取組織的總RNA，通過RT-PCR法檢測NFBD1基因在鼻咽癌組織中的mRNA表達水平；病理科取確診鼻咽癌組織的石蠟切片，及收集非鼻咽癌組織的處理后的石蠟切片，免疫組化法檢測NFBD1蛋白的表達水平及NFBD1蛋白在鼻咽癌細胞內(nèi)的分布情況。培養(yǎng)鼻咽癌細胞株HNE-1和CNE-1，將NFBD1siRNA導入鼻咽癌細胞株，分別采用RT-PCR和Western-blot法從mRNA水平及蛋白質(zhì)水平檢測siRNA對鼻咽癌細胞株中NFBD1表達的抑制效果；NFBD1表達受抑制后，給予化療藥（順鉑）處理各組細胞，24小時后MTT法分析鼻咽癌細胞對化療藥物敏感性的變化。 結(jié)果：鼻咽癌組織中NFBD1基因的RNA及蛋白質(zhì)表達水平均明顯高于正常的鼻咽部組織，鼻咽癌組織中NFBD1蛋白在胞核與胞質(zhì)均有表達，而在正常鼻咽部組織中，NFBD1蛋白主要分布于細胞質(zhì)中，胞核中表達量低。NFBD1siRNA轉(zhuǎn)染鼻咽癌細胞后，NFBD1的表達水平明顯降低；MTT結(jié)果分析表明，，NFBD1表達受抑制后，能顯著提高鼻咽癌細胞株對化療藥物的敏感性（P0.05）。 結(jié)論：NFBD1在鼻咽癌細胞中呈現(xiàn)高表達，其蛋白主要分布于胞核與細胞漿中，細胞DNA受到損失后，該蛋白向胞核移動。通鼻咽癌細胞中NFBD1表達受抑制后，可顯著提高鼻咽癌細胞株對化療藥物順鉑的敏感性。
[Abstract]:Aim: to investigate the expression of mRNA and protein in nasopharyngeal carcinoma (NPC) and the distribution of protein in nasopharyngeal carcinoma (NPC) tissues, and to study the effect of siRNA mediated inhibition of NFBD1 expression on the chemosensitivity of nasopharyngeal carcinoma (NPC) cell line.Methods: the total RNAs of nasopharyngeal carcinoma (NPC) and normal nasopharyngeal tissues were collected. The expression of NFBD1 gene in nasopharyngeal carcinoma (NPC) was detected by RT-PCR method, and the paraffin sections of nasopharyngeal carcinoma (NPC) tissues were obtained by pathology.The expression of NFBD1 protein and the distribution of NFBD1 protein in nasopharyngeal carcinoma cells were detected by immunohistochemistry.Nasopharyngeal carcinoma cell lines HNE-1 and CNE-1 were cultured, and NFBD1siRNA was introduced into nasopharyngeal carcinoma cell lines. The inhibitory effect of siRNA on NFBD1 expression in nasopharyngeal carcinoma cell lines was detected by RT-PCR and Western-blot from mRNA level and protein level respectively.After treated with cisplatin for 24 hours, the sensitivity of nasopharyngeal carcinoma cells to chemotherapeutic agents was analyzed by MTT.Results: the expression of RNA and protein of NFBD1 gene in nasopharyngeal carcinoma was significantly higher than that in normal nasopharyngeal tissue, and NFBD1 protein was expressed in nucleus and cytoplasm of nasopharyngeal carcinoma.However, in normal nasopharyngeal tissues, the expression of NFBD1 protein was mainly distributed in the cytoplasm, and the expression level of NFBD1 was significantly decreased after transfection of NFBD1 siRNA into nasopharyngeal carcinoma cells. The results of MTT analysis showed that the expression of NFBD1 was inhibited after transfection of NFBD1 siRNA into nasopharyngeal carcinoma cells.The sensitivity of nasopharyngeal carcinoma cell line to chemotherapeutic agents was significantly increased (P 0.05).Conclusion the protein of the protein mainly distributes in the nucleus and cytoplasm of nasopharyngeal carcinoma cells. After the loss of DNA, the protein moves to the nucleus of nasopharyngeal carcinoma cells.Inhibition of NFBD1 expression in NPC cells significantly increased the sensitivity of NPC cell lines to cisplatin.
【學位授予單位】：重慶醫(yī)科大學
【學位級別】：碩士
【學位授予年份】：2012
【分類號】：R739.63

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相關(guān)期刊論文 前1條

1 卜友泉;楊正梅;蘭歡;鎮(zhèn)磊;劉革力;宋方洲;;siRNA介導的NFBD1表達沉默對HeLa細胞增殖與凋亡的影響[J];中國生物化學與分子生物學報;2008年10期

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