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目的 :通过转导EB病毒 (EBV)转录的核内小RNA (EBERs)探讨其对舌癌细胞株OSC 19细胞生长特性的影响。方法 :EBERs基因编码区序列是利用聚合酶链反应 (PCR)技术从EB病毒阳性的鼻咽癌 (NPC)细胞株NPC KT细胞中扩增出。为了在细胞内大量表达EBERs,利用分子生物学技术制作了EBERs片段的 4次重复排列 4EBERs和 10次重复排列 10EBERs,分别亚克隆到 pCEP4载体质粒并转染OSC 19细胞。利用North ernBlot法确定细胞内EBERs的表达后 ,对EBERs转染后的OSC 19细胞在培养中的生长特性、软琼脂中的细胞克隆的形成进行了观察。结果 :在培养中大量表达EBERs的OSC 19细胞之间丧失了细胞间的相互连接 ,细胞呈纺锤状 ,并在软琼脂中形成大量细胞克隆。而非表达EBERs的OSC 19细胞没有发现细胞生长特性的改变 ,在软琼脂中没有形成大量细胞克隆。结论 :EBERs的大量表达可以导致OSC 19细胞的分散活性和基质非依赖性 ,提示EBERs在上皮细胞株OSC 19细胞中有致癌作用 ;大量表达EBERs的OSC 19细胞为进一步研究EBERs的作用提供了一个有用的上皮细胞模型。
OBJECTIVE: To investigate the effect of Epstein-Barr virus (EBV) on the growth of human tongue squamous carcinoma cell line OSC 19 by transduction of EBVs. Methods: EBERs gene coding region sequences were amplified from Epstein-Barr virus-positive nasopharyngeal carcinoma (NPC) NPC KT cells by polymerase chain reaction (PCR). In order to express large numbers of EBERs in cells, four EBERs fragment 4EBs and 10EBs were prepared by molecular biology techniques and subcloned into pCEP4 vector and transfected into OSC 19 cells respectively. After determining the expression of intracellular EBERs by North ernBlot method, the growth characteristics of cultured OSC 19 cells after EBERs transfection and the formation of cell colonies in soft agar were observed. Results: Intercellular connections were lost between OSC 19 cells that abundantly expressed EBERs in culture, spindle cells and formed large numbers of cell clones in soft agar. No change of cell growth was observed in OSC 19 cells that did not express EBERs, and a large number of cell clones were not formed in soft agar. Conclusion: The overexpression of EBERs can lead to the decentralization activity and matrix-independent activity of OSC 19 cells, suggesting that EBERs have carcinogenicity in epithelial cell line OSC 19; OSC 19 cells expressing EBERs in large quantities provide a further study of the role of EBERs Useful epithelial models.