In vivo silencing of Reptin blocks the progression of human hepatocellular carcinoma in xenografts and is associated with replicative senescence
In vivo silencing of Reptin blocks the progression of human hepatocellular carcinoma in xenografts and is associated with replicative senescence
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DOI：
10.1016/j.jhep.2009.12.029
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发表时间：
2010-05-01
影响因子：
25.7
通讯作者：
Rosenbaum, Jean
中科院分区：
文献类型：
--
作者：
Menard, Ludovic;Taras, Daniele;Rosenbaum, Jean
背景和目标：我们先前发现Reptin在肝细胞癌（HCC）中过表达，并且用siRNA体外耗尽Reptin导致HCC细胞生长停滞和凋亡。在这里，我们问是否在体内靶向Reptin在建立tumors.Methods的治疗效果：我们使用慢病毒载体构建HuH 7和Hep 3B细胞系与强力霉素（Dox）依赖性表达Reptin（R2）或对照shRNA（GL 2）。将细胞皮下注射到免疫缺陷小鼠中，当肿瘤体积达到250 mm 3时给予Dox（3）。结果：在体外，GL 2- Dox、GL 2 + Dox和R2 - Dox细胞的生长难以区分，而R2 + Dox细胞在Dox处理后4天停止生长。生长减少与细胞凋亡增加和复制衰老的证据有关，如酸性β-半乳糖苷酶染色和衰老相关异染色质灶的存在所示。在异种移植小鼠中，R2 + Dox肿瘤生长停滞，甚至随着长期治疗而消退，而GL 2- Dox、GL 2 + Dox和R2 - Dox肿瘤则稳定进展。肿瘤进展的阻断与衰老的诱导和细胞增殖减少有关。结论：在体内Reptin耗竭导致肿瘤生长停滞。Reptin可能是HCC治疗的一个有价值的靶点。(C)2010年欧洲肝脏研究协会。Elsevier B. V.出版，保留所有权利。
Background & Aims: We previously showed that Reptin is overexpressed in hepatocellular carcinoma (HCC), and that in vitro depletion of Reptin with siRNAs led to HCC cell growth arrest and apoptosis. Here, we asked whether in vivo targeting of Reptin in established tumours had a therapeutic effect.Methods: We used lentiviral vectors to construct HuH7 and Hep3B cell lines with doxycycline (Dox)-dependent expression of Reptin (R2) or control shRNA (GL2). Cells were injected subcutaneously into immunodeficient mice, and Dox was given when tumours reached a volume of 250 mm(3).Results: In vitro, the growth of GL2 - Dox, GL2 + Dox, and R2 - Dox cells was undistinguishable whereas that of R2 + Dox cells stopped 4 days after Dox treatment. The growth decrease was associated with increased apoptosis, and evidence of replicative senescence, as shown by staining for acid p-galactosidase and the presence of senescence-associated heterochromatin foci. In xenografted mice, R2 + Dox tumour growth stagnated or even regressed with prolonged treatment in contrast with the GL2 - Dox, GL2 + Dox, and R2 - Dox tumours that progressed steadily. The blockage of tumour progression was associated with the induction of senescence and reduced cell proliferation.Conclusions: In vivo Reptin depletion leads to tumour growth arrest. Reptin may prove a valuable target in HCC. (C) 2010 European Association for the Study of the Liver. Published by Elsevier B.V. All rights reserved.