%0 Journal Article %A Ying Gan %A Chenghua Cui %A Shengyan Xiang %A Baozhen Zhang %A Dajun Deng %T Effects of P16 DNA Methylation on Proliferation, Senescence, and Lifespan of Human Fibroblasts %D 2018 %R 10.1101/405407 %J bioRxiv %P 405407 %X The aim is to study the effects of P16 DNA methylation on lifespan of normal cells. An expression-controllable pTRIPZ vector expressing P26-specific zinc finger binding protein-based methyltransferase (P16-Dnmt) was used to induce P16 methylation in primary CCD-I8C0 fibroblasts via stable transfection. Long-term dynamic IncuCyte analysis showed that CCD-I8C0 fibroblasts expressing baseline P16-Dnmt continued proliferating until passage-26 in the 53th post-transfection week, while vector control cells stopped proliferating at passage-6 and completely died 2 weeks later. The proliferation rate of baseline P16-Dnmt cells was significantly higher than that of vector control cells. The proportion of P-galactosidase-positive staining cells was significantly decreased in baseline P16-Dnmt cells compared to vector control cells. The P16 expression was lost in baseline P16-Dnmt cells at and after passage-6. The average telomere length in baseline P16-Dnmt cells also gradually decreased. In conclusion, P16 methylation could prevent senescence, promote proliferation, and expand lifespan of human fibroblasts, which may play a role in cancer development.Summary A zinc finger protein-based DNA methyltransferase (P16-Dnmt) expressed at the baseline level could specifically methylate P16 promoter CpG islands. P16 methylation induced by baseline P16-Dnmt could significantly prevent senescence, promote proliferation, and expand lifespan of primary human fibroblasts.DHPLCdenatured high performance liquid chromatographyDnmtDNA methyltransferaseP16-DnmtP26-specific ZFP-DnmtTSStranscription start siteZFPzinc finger protein %U https://www.biorxiv.org/content/biorxiv/early/2018/08/30/405407.full.pdf