Primary human cervical carcinoma cells require human papillomavirus E6 and E7 expression for ongoing proliferation.

Primary human cervical carcinoma cells require human papillomavirus E6 and E7 expression for ongoing proliferation.
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DOI:
10.1016/j.virol.2011.10.012
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发表时间:
2012-01-05
期刊:
影响因子:
3.7
通讯作者:
DiMaio D
DiMaio D
中科院分区:
医学3区
文献类型:
--
作者:
Magaldi TG;Almstead LL;Bellone S;Prevatt EG;Santin AD;DiMaio D

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在已建立的宫颈癌细胞系中,人乳头瘤病毒(HPV)E6和E7癌基因的抑制由于细胞肿瘤抑制途径的重新激活而导致衰老。在这里,我们确定是否持续表达的HPV 16或HPV 18癌基因的增殖所需的原代人宫颈癌细胞在无血清条件下,在低传代次数后,从患者中分离。我们使用表达牛乳头瘤病毒E2蛋白的SV40病毒载体来抑制这些细胞中的E6和E7。为了实现有效的SV40感染和E2基因递送,我们首先将原代宫颈癌细胞与神经节苷脂GM1孵育,神经节苷脂GM1是SV40限制在这些细胞中的细胞表面受体。HPV在原代宫颈癌细胞中的抑制导致它们经历衰老,但E2蛋白对HPV阴性的原代细胞几乎没有影响。这些数据表明,E6和E7依赖性是人宫颈癌细胞的固有特性。
Repression of human papillomavirus (HPV) E6 and E7 oncogenes in established cervical carcinoma cell lines causes senescence due to reactivation of cellular tumor suppressor pathways. Here, we determined whether ongoing expression of HPV16 or HPV18 oncogenes is required for the proliferation of primary human cervical carcinoma cells in serum-free conditions at low passage number after isolation from patients. We used an SV40 viral vector expressing the bovine papillomavirus E2 protein to repress E6 and E7 in these cells. To enable efficient SV40 infection and E2 gene delivery, we first incubated the primary cervical cancer cells with the ganglioside GM1, a cell-surface receptor for SV40 limiting in these cells. Repression of HPV in primary cervical carcinoma cells caused them to undergo senescence, but the E2 protein had little effect on HPV-negative primary cells. These data suggest that E6 and E7 dependence is an inherent property of human cervical cancer cells.
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