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目的:了解RPMS1基因在鼻咽癌组织中的表达特点,并明确该基因在上皮细胞中表达的亚细胞区域,为进一步研究其生物学功能提供依据.方法:提取鼻咽癌组织及各细胞系的DNA和总RNA,以巢式PCR扩增W片段来检测Ep-ste in-Barr病毒(EB病毒)的感染情况,RT-PCR检测RPMS1基因在鼻咽癌组织、细胞株中的表达,并克隆入pEGFP-C2真核表达载体,通过脂质体转染技术将其介导入人胚肾上皮(HEK293)细胞,以激光共聚焦显微镜观察其蛋白表达的亚细胞区域.结果:除B JAB细胞株外,所有标本中均检测到EB病毒W片段;RPMS1 mRNA在鼻咽癌组织中的表达率为83.3%(45/54),在鼻咽非癌组织中则仅为4%(1/25),二者有显著性差异(P<0.001),鼻咽癌外周血淋巴细胞中未检测到RPMS1表达,RPMS1在SUNE1细胞中的表达高于Raji细胞,但不表达于B95.8细胞;RPMS1可稳定表达于HEK293细胞,其编码蛋白的表达以胞核为主.结论:EB病毒RPMS1基因在高发区鼻咽癌组织中存在高水平转录,在体外可稳定表达于上皮细胞的胞核,提示RPMS1编码蛋白可能作为核转录因子通过调控细胞增殖相关的信息传递而参与上皮癌变过程.
OBJECTIVE: To investigate the expression of RPMS1 gene in nasopharyngeal carcinoma (NPC) and to identify the subcellular region of the gene in epithelial cells for further study of its biological function.Methods: Nasopharyngeal carcinoma tissues and cell lines The DNA and total RNA were amplified by nested PCR to detect the infection of Ep-ste in-Barr virus (EBV). The expression of RPMS1 gene in nasopharyngeal carcinoma tissues and cell lines was detected by RT-PCR And then cloned into pEGFP-C2 eukaryotic expression vector and transfected into human embryonic kidney epithelial (HEK293) cells by lipofectamine 2000. The expression of subcellular domain was observed by laser confocal microscopy.Results: The EBV W fragment was detected in all the samples except for the cell lines. The expression of RPMS1 mRNA in nasopharyngeal carcinoma was 83.3% (45/54) and only 4% in nasopharyngeal non-cancerous tissues (1 / (P <0.001). No RPMS1 expression was detected in peripheral blood lymphocytes of NPC patients. The expression of RPMS1 in SUNE1 cells was higher than that in Raji cells but not in B95.8 cells. RPMS1 was stably expressed in HEK293 cells and the expression of its encoded protein was mainly nucleus.Conclusion: Nasopharyngeal carcinoma in the region of the high presence of transcription, stably expressed in the nucleus of epithelial cells in vitro, suggesting that RPMS1 encoded protein may act as a nuclear transcription factor by regulating cell proliferation-related information transmission involved in epithelial carcinogenesis.