NFBD1在鼻咽癌组织中的表达及抑制NFBD1的表达对鼻咽癌细胞化疗敏感性影响的研究

发布时间：2018-04-09 23:44

本文选题：NFBD1　切入点：鼻咽癌　出处：《重庆医科大学》2012年硕士论文

【摘要】：目的：了解NFBD1在鼻咽癌组织中mRNA及蛋白质的表达水平，及蛋白在细胞内的分布，并研究siRNA介导的鼻咽癌细胞株中NFBD1的表达抑制后，对鼻咽癌细胞株化疗敏感性的影响。方法：收集临床已确诊的鼻咽癌组织与正常鼻咽部组织，提取组织的总RNA，通过RT-PCR法检测NFBD1基因在鼻咽癌组织中的mRNA表达水平；病理科取确诊鼻咽癌组织的石蜡切片，及收集非鼻咽癌组织的处理后的石蜡切片，免疫组化法检测NFBD1蛋白的表达水平及NFBD1蛋白在鼻咽癌细胞内的分布情况。培养鼻咽癌细胞株HNE-1和CNE-1，将NFBD1siRNA导入鼻咽癌细胞株，分别采用RT-PCR和Western-blot法从mRNA水平及蛋白质水平检测siRNA对鼻咽癌细胞株中NFBD1表达的抑制效果；NFBD1表达受抑制后，给予化疗药（顺铂）处理各组细胞，24小时后MTT法分析鼻咽癌细胞对化疗药物敏感性的变化。结果：鼻咽癌组织中NFBD1基因的RNA及蛋白质表达水平均明显高于正常的鼻咽部组织，鼻咽癌组织中NFBD1蛋白在胞核与胞质均有表达，而在正常鼻咽部组织中，NFBD1蛋白主要分布于细胞质中，胞核中表达量低。NFBD1siRNA转染鼻咽癌细胞后，NFBD1的表达水平明显降低；MTT结果分析表明，，NFBD1表达受抑制后，能显著提高鼻咽癌细胞株对化疗药物的敏感性（P0.05）。结论：NFBD1在鼻咽癌细胞中呈现高表达，其蛋白主要分布于胞核与细胞浆中，细胞DNA受到损失后，该蛋白向胞核移动。通鼻咽癌细胞中NFBD1表达受抑制后，可显著提高鼻咽癌细胞株对化疗药物顺铂的敏感性。
[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.
【学位授予单位】：重庆医科大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R739.63

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