Down-regulation of GEP100 causes increase in E-cadherin levels and inhibits pancreatic cancer cell invasion.
Down-regulation of GEP100 causes increase in E-cadherin levels and inhibits pancreatic cancer cell invasion.
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DOI：
10.1371/journal.pone.0037854
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发表时间：
2012
期刊：
影响因子：
3.7
通讯作者：
Jia LT
中科院分区：
文献类型：
--
作者：
Xie CG;Wei SM;Chen JM;Xu XF;Cai JT;Chen QY;Jia LT
侵袭和转移是胰腺癌死亡的主要原因，识别特异性参与肿瘤侵袭的信号分子具有重要意义。本研究旨在探讨GEP 100在胰腺癌细胞侵袭转移中的作用及其分子机制。将GEP 100 shRNA载体转染PaTu 8988细胞，建立稳定的GEP 100敲低细胞系，并经嘌呤霉素筛选。通过qRT-PCR和Western blot检测基因表达。采用Matrigel侵袭实验检测癌细胞的体外侵袭能力。通过脾注射指定的细胞系，然后切除脾来确定体内肝转移。免疫荧光法检测E-cadherin在细胞内的定位。我们发现GEP 100蛋白的表达水平与一组6种不同的人胰腺癌细胞系的侵袭能力密切相关。通过Matrigel侵袭测定，PaTu 8988细胞中GEP 100的下调显著降低侵袭活性，而不影响迁移、侵袭和存活力。GEP 100 cDNA的过表达挽救了被抑制的侵袭活性。体内实验表明，GEP 100稳定敲减的PaTu 8988细胞肝转移能力明显降低。此外，上皮样形态学变化，模仿间充质上皮转化（MET）诱导GEP 100下调。E-cadherin蛋白表达增加2-3倍，并向细胞间重新分布，而E-cadherin mRNA表达无明显变化。出乎意料的是，Slug的mRNA通过GEP 100敲低而增加。这些结果为GEP 100通过调节E-cadherin的表达和MET的过程在胰腺癌侵袭中发挥重要作用提供了重要证据，提示其有可能成为胰腺癌潜在的治疗靶点。
Invasion and metastasis are major reasons for pancreatic cancer death and identifying signaling molecules that are specifically used in tumor invasion is of great significance. The purpose of this study was to elucidate the role of GEP100 in pancreatic cancer cell invasion and metastasis and the corresponding molecular mechanism. Stable cell lines with GEP100 knocked-down were established by transfecting GEP100 shRNA vector into PaTu8988 cells and selected by puromycin. qRT-PCR and Western blot were performed to detect gene expression. Matrigel-invasion assay was used to detect cancer cell invasion in vitro. Liver metastasis in vivo was determined by splenic injection of indicated cell lines followed by spleen resection. Immunofluorescence study was used to detect the intracellular localization of E-cadherin. We found that the expression level of GEP100 protein was closely related to the invasive ability of a panel of 6 different human pancreatic cancer cell lines. Down-regulation of GEP100 in PaTu8988 cells significantly decreased invasive activity by Matrigel invasion assay, without affecting migration, invasion and viability. The inhibited invasive activity was rescued by over-expression of GEP100 cDNA. In vivo study showed that liver metastasis was significantly decreased in the PaTu8988 cells with GEP100 stably knocked-down. In addition, an epithelial-like morphological change, mimicking a mesenchymal to epithelial transition (MET) was induced by GEP100 down-regulation. The expression of E-cadherin protein was increased 2–3 folds accompanied by its redistribution to the cell-cell contacts, while no obvious changes were observed for E-cadherin mRNA. Unexpectedly, the mRNA of Slug was increased by GEP100 knock-down. These findings provided important evidence that GEP100 plays a significant role in pancreatic cancer invasion through regulating the expression of E-cadherin and the process of MET, indicating the possibility of it becoming a potential therapeutic target against pancreatic cancer.
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影响因子：
254.7
作者：
Jemal, Ahmedin;Siegel, Rebecca;Thun, Michael J.
通讯作者：
Thun, Michael J.
影响因子：
3.7
作者：
Hashimoto A;Hashimoto S;Ando R;Noda K;Ogawa E;Kotani H;Hirose M;Menju T;Morishige M;Manabe T;Toda Y;Ishida S;Sabe H
通讯作者：
Sabe H
影响因子：
37.3
作者：
Keleg, Shereen;Buchler, Peter;Ludwig, Roman;Buchler, Markus W;Friess, Helmut
通讯作者：
Friess, Helmut
影响因子：
2.9
作者：
Deer EL;González-Hernández J;Coursen JD;Shea JE;Ngatia J;Scaife CL;Firpo MA;Mulvihill SJ
通讯作者：
Mulvihill SJ
影响因子：
4.5
作者：
Dunphy, Jillian L.;Ye, Keqiang;Casanova, James E.
通讯作者：
Casanova, James E.