[Transcriptional inhibition of human papilloma virus in cervical carcinoma cells reactivates functions of the tumor suppressor p53].
[Transcriptional inhibition of human papilloma virus in cervical carcinoma cells reactivates functions of the tumor suppressor p53].
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发表时间：
2007-05
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通讯作者：
D. V. Kochetkov;Il'inskaia Gv;P. Komarov;E. Strom;L. Agapova;A. V. Ivanov;A. Budanov;Elena I. Frolova;P. Chumakov
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作者：
D. V. Kochetkov;Il'inskaia Gv;P. Komarov;E. Strom;L. Agapova;A. V. Ivanov;A. Budanov;Elena I. Frolova;P. Chumakov
人类大多数恶性疾病都伴随着肿瘤抑制因子 p53 的失活。肿瘤中p53功能的恢复会导致癌细胞死亡，这可用于癌症治疗。在宫颈癌中，人乳头瘤病毒E6基因的产物促进蛋白酶体系统中p53的加速降解。因此，在宫颈癌细胞中重新激活 p53 的方法之一可能是使用抑制病毒蛋白功能的小分子。通过使用人宫颈癌细胞（携带人乳头状瘤病毒 18 型，HPV-18 的 HeLa 细胞系）作为测试系统，引入了在 p53 依赖性启动子控制下表达 β-半乳糖苷酶的报告构建体，我们对小分子文库进行了筛选，以选择能够重新激活 p53 转录活性的小分子。然后，我们表征了两种最活跃的化合物在 p53 依赖性信号通路状态不同的细胞系中的作用。这两种化合物在表达 HPV-18 的细胞系（较小程度上是 HPV-16）中引起 p53 的特异性激活，并且不会对对照 p53 阴性细胞或 p53 途径未中断的细胞产生任何影响。宫颈癌细胞中 p53 的激活伴随着 p53 依赖性基因 CDKN1 (p21) 的诱导、增殖的抑制以及细胞凋亡的诱导。这两种化合物都能够深度抑制 HPV 基因组的转录，这显然是 E6 蛋白表达下降导致 p53 重新激活的原因。观察到的对正常细胞的低毒性允许将这些化合物视为未来抗癌药物的原型。
Inactivation of tumor suppressor p53 accompanies the majority of malignant diseases in humans. Restoration of p53 functions in tumor results in death of cancer cells, which can be used in cancer therapy. In cervical cancer a product of E6 gene of the human papilloma virus promotes accelerated degradation of p53 in proteasome system. Therefore, one of the approaches to reactivation of p53 in cervical carcinoma cells could be the use of small molecules that inhibit functions of viral proteins. By using as a test system human cervical carcinoma cells (HeLa cell line bearing human papilloma virus type 18, HPV-18) with introduced reporter construct that expresses beta-galactosidase under control of a p53-dependent promoter we carried out screening of a library of small molecules to select small molecules capable of reactivating transcriptional activity of p53. We then characterized the effects of two most active compounds in cell lines that differ in the status of p53-dependent signaling pathway. Both of the compounds caused specific activation of p53 in the cell lines expressing HPV-18, to a lesser extent--HPV-16, and do not cause any effect in control p53 negative cells, or in the cells with undisrupted p53 pathway. Activation of p53 in cervical carcinoma cells was accompanied by the induction of the p53-dependent gene CDKN1 (p21), by inhibition of proliferation, and by the induction of apoptosis. Both of the compounds were capable of deep inhibition of transcription from the HPV genome, which apparently was the cause for p53 reactivation in response to decreased expression of the E6 protein. The observed low toxicity for normal cells allows considering these chemical compounds as prototypes for future anticancer drugs.