Increased expression of the large GTPase dynamin 2 potentiates metastatic migration and invasion of pancreatic ductal carcinoma.
Increased expression of the large GTPase dynamin 2 potentiates metastatic migration and invasion of pancreatic ductal carcinoma.
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DOI：
10.1038/onc.2011.329
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发表时间：
2012-03-08
期刊：
影响因子：
8
通讯作者：
McNiven, M. A.
中科院分区：
文献类型：
--
作者：
Eppinga, R. D.;Krueger, E. W.;Weller, S. G.;Zhang, L.;Cao, H.;McNiven, M. A.
胰腺导管肿瘤侵犯局部实质并转移到远处器官。Src介导的酪氨酸激酶信号通路促进胰腺导管腺癌（PDAC）转移，尽管支持这种侵袭性过程的分子机制知之甚少，并代表重要和新的治疗靶点。大的GT3激酶Dynamin 2（Dyn 2），一种Src激酶底物，调节膜细胞骨架动力学，尽管尚未确定这种机械酶是否有助于肿瘤细胞迁移和侵袭。因此，本研究的目的是测试Dyn 2是否在人胰腺肿瘤中上调，并使用体外测定和裸鼠模型确定其在细胞迁移和转移侵袭中的作用。组织学分析显示，85名受试患者中有81%在PDAC组织中Dyn 2升高。为了测试Dyn 2过表达是否改变人胰腺肿瘤细胞的转移特性，产生过表达野生型Dyn 2或已知减弱Dyn 2功能的磷酸化缺陷突变体Dyn 2 Y（231/597）F的BxPC-3细胞的稳定克隆，并分析其迁移能力。重要的是，与表达Dyn 2 Y（231/597）F或单独载体的细胞相比，表达2-3倍水平的Dyn 2的肿瘤细胞以两倍的速率突出板状伪足，在玻璃上和通过transwell室（通过过滤器的细胞多2-3倍）迁移更快（180%）和更远（2.5倍的净距离）。此外，与DMSO处理的细胞相比，siRNA介导的Dyn 2和发动蛋白抑制剂MiTMAB和Dynasore的消耗显著降低了transwell测定中的细胞迁移（>66%）、伤口愈合（>75%）和侵袭（>95%）。为了测试增加的Dyn 2表达所赋予的转移潜力，将BxPC-3克隆细胞系原位植入裸鼠的胰腺中。与仅表达Dyn 2 Y（231/597）F或载体的细胞相比，表达Dyn 2-GFP的细胞在大的远端肿瘤中表现出3倍的增加。最后，来自人类患者的胰腺转移瘤的组织学分析显示，Dyn 2在60%的检查的转移性肿瘤中上调。这些发现是第一个牵连发动蛋白在任何肿瘤的条件，并直接证明了这种机械酶在侵入性细胞迁移的作用。
Pancreatic ductal tumors invade local parenchyma and metastasize to distant organs. Src-mediated tyrosine kinase signaling pathways promote pancreatic ductal adenocarcinoma (PDAC) metastasis, though the molecular mechanisms supporting this invasive process are poorly understood and represent important and novel therapeutic targets. The large GTPase Dynamin2 (Dyn2), a Src-kinase substrate, regulates membrane-cytoskeletal dynamics although it is yet to be defined if this mechanoenzyme contributes to tumor cell migration and invasion. Therefore the goal of this study was to test if Dyn2 is upregulated in human pancreatic tumors and to define its role in cell migration and metastatic invasion using in vitro assays and nude mouse models. Histological analysis showed that 81% of the 85 patients tested had elevated Dyn2 in PDAC tissue. To test if Dyn2 overexpression alters metastatic properties of human pancreatic tumor cells, stable clones of BxPC-3 cells overexpressing either wild-type Dyn2 or a phosphorylation-deficient mutant Dyn2Y(231/597)F known to attenuate Dyn2 function, were generated and analyzed for migratory capacity. Importantly, tumor cells expressing 2-3 fold levels of Dyn2 protruded lamellipodia at twice the rate, migrated faster (180%) and farther (2.5-fold greater net distance) on glass and through transwell chambers (2-3 fold more cells through the filter) compared to cells expressing Dyn2Y(231/597)F or vector alone. Further, siRNA-mediated depletion of Dyn2 and dynamin inhibitors MiTMAB and Dynasore significantly reduced cell migration (>66%), wound healing (>75%) and invasion in transwell assays (>95%) compared to DMSO treated cells. To test the metastatic potential conferred by increased Dyn2 expression, the BxPC-3 clonal cell lines were implanted orthotopically into the pancreas of nude mice. Cells expressing Dyn2-GFP exhibited a 3-fold increase in large distal tumors compared to cells expressing Dyn2Y(231/597)F or vector alone. Finally, histological analysis of pancreatic metastases from human patients revealed that Dyn2 is upregulated in 60% of metastatic tumors examined. These findings are the first to implicate dynamin in any neoplastic condition and to directly demonstrate a role for this mechanoenzyme in invasive cell migration.
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影响因子：
4.8
作者：
Ahn, S;Maudsley, S;Daaka, Y
通讯作者：
Daaka, Y
DOI：
10.1083/jcb.200904054
发表时间：
2009-11-30
期刊：
The Journal of cell biology
影响因子：
--
作者：
Ezratty EJ;Bertaux C;Marcantonio EE;Gundersen GG
通讯作者：
Gundersen GG
影响因子：
21.3
作者：
Ezratty, EJ;Partridge, MA;Gundersen, GG
通讯作者：
Gundersen, GG
影响因子：
3.5
作者：
Chao, Wei-Ting;Kunz, Jeannette
通讯作者：
Kunz, Jeannette
影响因子：
4.8
作者：
Ahn, S;Kim, J;Daaka, Y
通讯作者：
Daaka, Y