Thompson D A, Belinsky G, Chang T H, Jones D L, Schlegel R, Münger K
Department of Pathology, Harvard Medical School, Boston, Massachusetts 02115, USA.
Oncogene. 1997 Dec 18;15(25):3025-35. doi: 10.1038/sj.onc.1201495.
The E6 and E7 proteins of the high risk human papillomaviruses (HPVs) are consistently expressed in HPV-positive cervical carcinomas. We investigated the ability of HPV-16 E6 and E7 to disrupt mitotic checkpoints in normal diploid human cells. Acute expression of HPV-16 E6, but not HPV-16 E7, decreased the fidelity of multiple checkpoints controlling entry into and exit from mitosis. After irradiation, nearly 50% of cells containing HPV-16 E6 readily entered mitosis as opposed to less than 10% of control cells. Consistent with this, asynchronous populations of cells expressing HPV-16 E6 had increased cdc2-associated histone H1 kinase activity relative to control populations. In addition, HPV-16 E6 increased sensitivity to chemically-induced S-phase premature mitosis and decreased mitotic spindle assembly checkpoint function relative to control populations. HPV-16 E6 mutants with a reduced ability to target p53 for degradation were unable to abrogate mitotic checkpoints, suggesting a possible mechanism by which HPV-16 E6 disrupts mitotic checkpoints. Expression of a mutant p53 gene yielded an intermediate phenotype relative to HPV-16 E6, generating moderate increases in sensitivity to chemically-induced S-phase PCC and mitotic spindle disruption and a heightened propensity to enter mitosis after irradiation.
高危型人乳头瘤病毒(HPV)的E6和E7蛋白在HPV阳性的宫颈癌中持续表达。我们研究了HPV-16 E6和E7破坏正常二倍体人类细胞有丝分裂检查点的能力。HPV-16 E6而非HPV-16 E7的急性表达降低了控制有丝分裂进出的多个检查点的保真度。照射后,近50%含有HPV-16 E6的细胞容易进入有丝分裂,而对照细胞不到10%。与此一致的是,相对于对照群体,表达HPV-16 E6的异步细胞群体具有增加的与cdc2相关的组蛋白H1激酶活性。此外,相对于对照群体,HPV-16 E6增加了对化学诱导的S期早熟有丝分裂的敏感性,并降低了有丝分裂纺锤体组装检查点功能。靶向p53进行降解能力降低的HPV-16 E6突变体无法消除有丝分裂检查点,这表明HPV-16 E6破坏有丝分裂检查点的一种可能机制。突变型p53基因的表达相对于HPV-16 E6产生了中间表型,对化学诱导的S期早熟染色体凝缩和有丝分裂纺锤体破坏的敏感性适度增加,并且在照射后进入有丝分裂的倾向增强。