人乳头瘤病毒16型长控制区上YY1结合位点的破坏可增强病毒诱导细胞永生化能力  

REMOVAL OF THE SPECIFIC BINDING SITES FOR YY1 IN HPV 16 LCR ENHANCES THE VIRAL IMMORTALIZATION CAPACITY ON PRIMARY KERATINOCYTES FROM FORESKIN

作　　者：董小平[1] 刘红[2] 周伟[1] 

机构地区：[1]中国预防医学科学院病毒学研究所,北京100052 [2]首都医科大学附属口腔医院,北京100050

出　　处：《病毒学报》1999年第3期216-224,共9页Chinese Journal of Virology

摘　　要：细胞转录调节因子 Ｙ Ｙ１ 可抑制人乳头瘤病毒１６ 型（ Ｈ Ｐ Ｖ １６） 癌基因启动子 Ｐ９７ 的活性， Ｙ Ｙ１ 位点的突变和缺失不仅可诱导 Ｐ９７ 活性增强而且可在全基因组内增强 Ｅ６ 癌基因转录，同时使病毒对啮齿类动物纤维细胞的转化能力增强。为了观测人乳头瘤病毒１６ 型长控制区（ Ｈ Ｐ Ｖ１６ Ｌ Ｃ Ｒ） 序列上 Ｙ Ｙ１ 蛋白特异性结合位点破坏在完整基因组范围内对人原代包皮角源细胞永生化能力的影响，将 Ｈ Ｐ Ｖ １６ Ｙ Ｙ１ 位点突变株和野毒株转染至人原代包皮角源细胞。筛选结果表明，突变株可诱导形成永生化细胞，永生化能力明显高于野毒株。对４ 株永生化细胞系 Ｄ Ｎ Ａ检测发现，均含有呈整合状态的 Ｈ Ｐ Ｖ １６ Ｄ Ｎ Ａ，其中３ 株的 Ｅ１／ Ｅ２ 区域有缺失。 Ｒ Ｎ Ａ 检测显示，４株细胞内均有 Ｅ６／ Ｅ７ ｍ Ｒ Ｎ Ａ 的转录。这表明， Ｈ Ｐ Ｖ １６ Ｌ Ｃ Ｒ 上 Ｙ Ｙ１ 蛋白特异性结合位点的破坏，可在完整基因组范围内增强病毒使人原代包皮角源细胞永生化的能力。Cellular transcription regulator YY1 represses the activity of the viral oncogene promoter P97. Mutations and deletions of the specific binding sites for YY1 could not only increase the P97 activity, but also elevate the transcription of viral E6 in the transient transfection and the transforming ability on the rodent fibroblasts in the context of the whole genome. In order to test the effect of removal of YY1 binding sites on the immortalization capacity on the human primary keratinocytes from foreskin, we transfected both wild-type and YY1 binding site mutated HPV 16 DNAs into the keratinocytes. The selecting results showed that the mutated HPV 16 DNAs were able to, whereas the wild-type HPV 16 DNA failed to, induce the immortalized clones. DNA analysis of 4 immortalized clones revealed that all the 4 clones contained the integrated HPV 16 DNAs with intact E6/E7 sequence and LCR region, while 3 of them had unintact E1 and /or E2 regions. Northern blot and primer extension tests confirmed that viral oncogenes E6 and E7 were consistently transcribed in all the 4 clones. All of these suggest that removal of YY1 binding sites can increase viral immortalization capacity, and might enhance the viral tumourigenicity.

关 键 词：人乳头瘤病毒 细胞转录 调节因子 永生化 癌基因 

分 类 号：R373.9[医药卫生—病原生物学]