Boccardo Enrique, Noya Francisco, Broker Thomas R, Chow Louise T, Villa Luisa L
Ludwig Institute for Cancer Research, 1509-010 São Paulo, SP, Brazil.
Virology. 2004 Oct 25;328(2):233-43. doi: 10.1016/j.virol.2004.07.026.
The proinflammatory cytokine tumor necrosis factor-alpha (TNF-alpha) inhibits normal keratinocytes proliferation. However, many human papillomavirus (HPV)-immortalized or transformed cell lines are resistant to TNF-alpha antiproliferative effect. The present study analyzes the effects of TNF-alpha on organotypic cultures of primary human keratinocytes (PHKs) that express HPV-18 oncogenes. Raft cultures prepared with PHKs acutely transfected with HPV-18 whole genome or infected with recombinant retroviruses containing only E6/E7 or E7 were treated with 2 nM TNF-alpha. While BrdU incorporation into basal/parabasal cells of normal PHKs cultures was markedly inhibited by TNF-alpha cultures transfected with HPV-18 whole genome showed proliferation in all cell strata. Furthermore, BrdU incorporation into cultures expressing E6/E7 or E7 was not significantly reduced, indicating that E7 alone confers partial resistance to TNF-alpha. Besides, TNF-alpha treatment did not alter p16ink4a, p21cip1, p27kip1, or cyclin E levels, but did reduce cyclin A and PCNA levels in sensitive cells.
促炎细胞因子肿瘤坏死因子-α（TNF-α）可抑制正常角质形成细胞的增殖。然而，许多人乳头瘤病毒（HPV）永生化或转化的细胞系对TNF-α的抗增殖作用具有抗性。本研究分析了TNF-α对表达HPV-18癌基因的原代人角质形成细胞（PHK）器官型培养物的影响。用HPV-18全基因组急性转染或感染仅含E6/E7或E7的重组逆转录病毒的PHK制备的筏式培养物，用2 nM TNF-α处理。虽然TNF-α显著抑制正常PHK培养物的基底/副基底细胞中BrdU的掺入，但转染HPV-18全基因组的培养物在所有细胞层均显示增殖。此外，掺入表达E6/E7或E7的培养物中的BrdU没有显著减少，表明单独的E7赋予对TNF-α的部分抗性。此外，TNF-α处理未改变p16ink4a、p21cip1、p27kip1或细胞周期蛋白E的水平，但确实降低了敏感细胞中细胞周期蛋白A和增殖细胞核抗原（PCNA）的水平。
