Amplification of chromosome 5p correlates with increased expression of Skp2 in HPV-immortalized keratinocytes
Amplification of chromosome 5p correlates with increased expression of Skp2 in HPV-immortalized keratinocytes
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DOI：
10.1038/sj.onc.1206296
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发表时间：
2003-04-24
期刊：
影响因子：
8
通讯作者：
Stanley, MA
中科院分区：
文献类型：
--
作者：
Dowen, SE;Neutze, DM;Stanley, MA
在体外，致癌的人乳头瘤病毒使原代生殖器角化细胞永活，有证据表明，这些细胞系是检测人乳头瘤病毒诱导的癌变的合适模型。早期体内研究和最近的CGH分析显示，晚期子宫颈癌（CaCx）中存在5p染色体扩增。在本研究中，通过M-FISH分析了一组已建立的cacx衍生细胞时间，以确定复发性核型异常。在携带HR（高危）HPV的13株CaCx细胞系中，观察到5p的扩增。5p被扩增的区域用人类波段特异性颜料确认。F-box蛋白Skp2存在于5p13中，其蛋白在许多晚期癌症中水平升高。携带hpv16的细胞系W12在体外传代时表现出进行性形态异常，最终形成侵袭性表型。利用Western blot和TaqMan定量PCR研究Skp2在这一过程中不同阶段的表达。传代中后期，5p作为同工染色体获得。Skp2的表达增加和其靶蛋白p27的表达减少与该细胞系传代增加相关。
The oncogenic HPVs immortalize primary genital keratinocytes in vitro and there is evidence that such lines represent suitable models to examine HPV-induced carcinogenesis. Early in vivo studies and more recent CGH analyses have revealed amplification of chromosome 5p in advanced stage carcinoma of the uterine cervix (CaCx). In the present study, a panel of established CaCx-derived cell tines were analysed by M-FISH to identify recurrent karyotypic abnormalities. Amplification of 5p was observed in I I of 13 CaCx cell lines harbouring HR (high-risk) HPV. The region of 5p undergoing amplification was confirmed using human band-specific paints. The F-box protein Skp2 is present at 5p13 and its protein is present at increased levels in many cancers of an advanced stage. The HPV16-harbouring cell line W12 shows progressive morphological abnormality with in vitro passage, culminating in an invasive phenotype. The expression of Skp2 at different stages of this progression was investigated utilizing Western blot and TaqMan quantitative PCR. At medium to late passage, gain of 5p as an isochromosome was observed. Increased expression of Skp2 and a reduction in the expression of its target p27 correlated with increasing passage in this line.