Role of SV40 integration site at chromosomal interval 1q21.1 in immortalized CRL2504 cells.
Role of SV40 integration site at chromosomal interval 1q21.1 in immortalized CRL2504 cells.
复制标题
永生化 CRL2504 细胞中染色体间隔 1q21.1 的 SV40 整合位点的作用。
DOI：
10.1158/0008-5472.can-09-1003
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发表时间：
2009
期刊：
影响因子：
11.2
通讯作者：
Athwal,RaghbirS
中科院分区：
文献类型：
--
作者：
Liu,Jinglan;Kaur,Gurpreet;Zhawar,VikramjitK;Zimonjic,DrazenB;Popescu,NicholasC;Kandpal,RajP;Athwal,RaghbirS
我们应用功能性基因转移策略来显示病毒整合位点在细胞永生化中的重要性。在病毒转化的人细胞中，SV40的大肿瘤抗原能够通过隔离肿瘤抑制蛋白pRB和p53来延长细胞寿命。虽然SV40大T抗原是必不可少的，但它不足以使细胞永生，这表明需要细胞基因的额外改变才能实现无限增殖。我们在这里表明，SV40整合对人类染色体间隔1q21.1的破坏可能是细胞永生的重要步骤。一个150 kb的细菌人工染色体（BAC）克隆，RP364B14，对应于CRL2504细胞的病毒整合位点的转移，恢复了它们的不朽表型。有趣的是，CRL2504细胞的BAC转移克隆显示出β-半乳糖苷酶活性显示的衰老特征或M30 CytoDEATH抗体阳性染色显示的凋亡特征。SV40在1q21.1位点整合在表皮分化复合体（EDC）基因附近，导致作为EDC一部分的聚丝蛋白（FLG）基因下调。FLGgene在BAC转移、衰老和凋亡克隆中表达升高。我们的研究结果表明，SV40对原生基因组序列的破坏可能通过调节染色质结构来改变衰老和凋亡相关基因的表达。这些研究表明，鉴定人类癌症病毒整合位点附近的基因可能有助于开发新的诊断和治疗策略。[癌症研究2009；69 (19): 7819 - 25)
We have applied a functional gene transfer strategy to show the importance of viral integration site in cellular immortalization. The large tumor antigen of SV40 is capable of extending the cellular life span by sequestering tumor suppressor proteins pRB and p53 in virus-transformed human cells. Although SV40 large T antigen is essential, it is not sufficient for cellular immortalization, suggesting that additional alterations in cellular genes are required to attain infinite proliferation. We show here that the disruption of human chromosomal interval at 1q21.1 by SV40 integration can be an essential step for cellular immortalization. The transfer of a 150-kb bacterial artificial chromosome (BAC) clone, RP364B14, corresponding to viral integration site in CRL2504 cells, reverted their immortal phenotype. Interestingly, the BAC transfer clones of CRL2504 cells displayed characteristics of either senescence as shown by β-galactosidase activity or apoptosis as revealed by positive staining with M30 CytoDEATH antibody. The SV40 integration at 1q21.1, in the vicinity of epidermal differentiation complex (EDC) genes, resulted in the down-regulation of thefilaggrin(FLG) gene that is part of theEDC.FLGgene expression was increased in BAC transfer senescent and apoptotic clones. Our results suggest that the disruption of native genomic sequence by SV40 may alter expression of genes involved in senescence and apoptosis by modulating chromatin structure. These studies imply that identification of genes located in the vicinity of viral integration sites in human cancers may be helpful in developing new diagnostic and therapeutic strategies. [Cancer Res 2009;69(19):7819–25]