HPV 16 YY1位点突变株诱导的永生化细胞系具有部分转化细胞特征

目的 检测ＨＰＶ １６ ＹＹ１位点突变株诱导的包皮角原细胞永生化细胞系的生物学特性。方法 提取永生化细胞株蛋白，以Ｗｅｓｔｅｒｎ ｂｌｏｔ检测细胞内源性ｐ５３蛋白含量；以ＴＲＡＰ法检测细胞中端粒体酶活性；将永生化细胞系接种于含有１０％ＦＣＳ的软琼脂培养基，观测细胞的软琼脂生长能力。结果 Ｗｅｓｔｅｒｎ ｂｌｏｔ检测结果显示４株永生化细胞系中ｐ５３均为阴性；端粒体酶活性均为阳性，并且随着细胞传代活性

The immortalized cell lines induced by hunan papillomavirus type 16 (HPVS 16) YY1 binding-site mu-tants have some characteristics of transformed cells.

Objective To study the biological characteristics of the immortalized cell lines of human keratinocytes from foreskin induced by human papillomavirus type 16 (HPV 16) YY1 binding-site mutants.Methods The cellular extracts of immortalized cell lines were prepared and the cellular endogenous p53 proteins were determined with Western blot.The cellular telomerase activities were analyzed with TRAP The immortalized cells were incubated in a soft-agarose medium containing 10% FCS and the anchorage independent growth abilities of the tested cell lines were evaluated. Results Western blot showed that the endogenous p53 protein in all 4 tested clones were undetectable.The telomerases were clearly identified within the 4 tested cell lines,and showed an increasing activity along with the passages of the cells.All the clones were unable to grow in the soft-agarose medium during the earlier passages,but three of them showed anchorage-independent-growth at the passages of 30-35.Conclusion The immortalized cell slines induced by the HPV 16 YY1 binding-site mutants have active telomerase activity and ability of anchorage-independent growth.