Sumoylation in p27kip1 via RanBP2 promotes cancer cell growth in cholangiocarcinoma cell line QBC939.
Sumoylation in p27kip1 via RanBP2 promotes cancer cell growth in cholangiocarcinoma cell line QBC939.
复制标题
RanBP2 对 p27kip1 的 Sumoylation 促进胆管癌细胞系 QBC939 中的癌细胞生长
DOI：
10.1186/s12867-017-0100-5
复制
发表时间：
2017-09-07
影响因子：
--
通讯作者：
Luo J
中科院分区：
文献类型：
--
作者：
Yang J;Liu Y;Wang B;Lan H;Liu Y;Chen F;Zhang J;Luo J
背景胆管细胞癌是一种5年生存率低、对常规治疗反应差的致命性疾病。此前，我们发现p27kip1核浆易位对胆管癌细胞系QBC939具有增殖潜能，这一过程是由CRM-1介导的。结果在本研究中，我们探讨了p27kip1在核浆转位中的作用及其与QBC939细胞增殖的关系。首先，我们确定K73是p27kip1中的苏莫化位点。通过利用质粒FLAG-p27kip1、HA-RanBP2、GST-RanBP2和His-p27kip1以及免疫沉淀实验，我们验证了p27kip1可以作为QBC939中RanBP2的总甲基化靶点。此外，我们还证实了RAMBP2的S在促进p27kip1核浆转位中的作用，并且发现RAMBP2的S功能依赖于RAMP-1。然而，在p27kip1易位过程中，K73R突变的p27kip1不能被CRM-1和RanBP2识别，提示在这一过程中，RanBP2和CRM-1需要通过K73位点对p27kip1进行总合作用。表型上，RanBP2或CRM-1的过表达可以部分挽救MTS和EDU实验中p27kip1过表达所带来的抗增殖作用。我们首次发现并验证了p27kip1上的K73总甲基化位点，该位点在p27kip1核浆转位过程中对RanBP2和CRM-1起关键作用。结论靶向抑制p27kip1总合作用有望成为根治胆管癌发生和复发的潜在治疗靶点。
BackgroundCholangiocarcinoma is one of the deadly disease with poor 5-year survival and poor response to conventional therapies. Previously, we found that p27kip1 nuclear-cytoplasmic translocation confers proliferation potential to cholangiocarcinoma cell line QBC939 and this process is mediated by crm-1. However, no other post-transcriptional regulation was found in this process including sumoylation in cholangiocarcinoma.ResultsIn this study, we explored the role of sumoylation in the nuclear-cytoplasmic translocation of p27kip1 and its involvement of QBC939 cells’ proliferation. First, we identified K73 as the sumoylation site in p27kip1. By utilizing plasmid flag-p27kip1, HA-RanBP2, GST-RanBP2 and His-p27kip1 and immunoprecipitation assay, we validated that p27kip1 can serve as the sumoylation target of RanBP2 in QBC939. Furthermore, we confirmed crm-1’s role in promoting nuclear-cytoplasmic translocation of p27kip1 and found that RanBP2’s function relies on crm-1. However, K73R mutated p27kip1 can’t be identified by crm-1 or RanBP2 in p27kip1 translocation process, suggesting sumoylation of p27kip1 via K73 site is necessary in this process by RanBP2 and crm-1. Phenotypically, the overexpression of either RanBP2 or crm-1 can partially rescue the anti-proliferative effect brought by p27kip1 overexpression in both the MTS and EdU assay. For the first time, we identified and validated the K73 sumoylation site in p27kip1, which is critical to RanBP2 and crm-1 in p27kip1 nuclear-cytoplasmic translocation process.ConclusionTaken together, targeted inhibition of sumoylation of p27kip1 may serve as a potentially potent therapeutic target in the eradication of cholangiocarcinoma development and relapses.
登录
查看更多内容
影响因子：
14.9
作者：
Xue, Yu;Zhou, Fengfeng;Fu, Chuanhai;Xu, Ying;Yao, Xuebiao
通讯作者：
Yao, Xuebiao
影响因子：
16.6
作者：
Bellail, Anita C.;Olson, Jeffrey J.;Hao, Chunhai
通讯作者：
Hao, Chunhai
影响因子：
11.8
作者：
Kaminsky, Rachel;Denison, Carilee;Bening-Abu-Shach, Ulrike;Chisholm, Andrew D.;Gygi, Steven P.;Broday, Limor
通讯作者：
Broday, Limor
影响因子：
5.3
作者：
Larrea, Michelle D.;Liang, Jiyong;Slingerland, Joyce M.
通讯作者：
Slingerland, Joyce M.
影响因子：
8.8
作者：
Ritho, Joan;Arold, Stefan T.;Yeh, Edward T. H.
通讯作者：
Yeh, Edward T. H.