Girdin interaction with vimentin induces EMT and promotes the growth and metastasis of pancreatic ductal adenocarcinoma

Wang,Wulin; Chen,Hao; Gao,Wenjie; Wang,Sheng; Wu,Kai; Lu,Chen; Luo,Xiagang; Li,Lianhong; Yu,Chunzhao

doi:10.3892/or.2020.7615

Girdin interaction with vimentin induces EMT and promotes the growth and metastasis of pancreatic ductal adenocarcinoma

Authors:
- Wulin Wang
- Hao Chen
- Wenjie Gao
- Sheng Wang
- Kai Wu
- Chen Lu
- Xiagang Luo
- Lianhong Li
- Chunzhao Yu
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Affiliations: Department of General Surgery, Second Affiliated Hospital, Nanjing Medical University, Nanjing, Jiangsu 210000, P.R. China, Department of Hepatobiliary and Pancreatic Surgery, The Affiliated Suqian Hospital of Xuzhou Medical University, Suqian, Jiangsu 223800, P.R. China, Department of Gastrointestinal Surgery, Second Affiliated Hospital of Changzhou, Nanjing Medical University, Changzhou, Jiangsu 213000, P.R. China
Published online on: May 19, 2020 https://doi.org/10.3892/or.2020.7615
Pages: 637-649
Copyright: © Wang et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Pancreatic ductal adenocarcinoma (PDAC) is a highly malignant cancer of the digestive tract that has a high potential for metastasis and a poor prognosis. Girdin was first reported in 2005 as an actin‑binding protein and was designated as Akt‑phosphorylation enhancer (APE); thus, Girdin has been revealed to have an important role in regulating cancer development. There is additional evidence indicating that Girdin is associated with cell proliferation, migration, invasion and survival in certain cancers. However, the potential mechanisms involving Girdin and mobility in pancreatic cancer have not been elucidated. In the present study, it was revealed that Girdin was highly expressed in pancreatic cancer tissue and was associated with tumor grade. The present study, to the best of our knowledge, is the first aimed at investigating the unknown role of Girdin in PDAC metastasis. A short hairpin RNA for Girdin (sh‑Girdin) was successfully constructed with recombinant adenoviral vectors to suppress the expression of Girdin in pancreatic cancer cell lines (PANC‑1 and BXPC‑3). The silencing efficiency of the Girdin shRNA was determined by RT‑qPCR and western blot analysis, and decreased Girdin expression in the cytoplasm was revealed by immunofluorescence detection. Then, sulforhodamine B (SRB) and colony formation assays were used to confirm that the knockdown of Girdin inhibited proliferation in vitro, and Transwell assays were used to examine the influence of Girdin knockdown on cellular mobility. Animal experiments also confirmed that silencing the expression of Girdin in pancreatic cancer cells inhibited the growth and metastasis of pancreatic cancer in vivo. Transforming growth factor‑β (TGF‑β) is a common inducer of epithelial‑mesenchymal transition (EMT) and can effectively induce EMT in PDAC. Notably, TGF‑β‑treated cells exhibited changes in the classic biological markers of EMT. The expression of E‑cadherin, a marker of the epithelial phenotype, increased, and the expression of N‑cadherin and vimentin, markers of the interstitial phenotype, decreased in response to sh‑Girdin. According to these experiments, Girdin may affect pancreatic cancer progression and development by interacting with vimentin. Therefore, there is evidence indicating that Girdin could be designated as a prognostic biological indicator and a candidate therapeutic target for pancreatic cancer.

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1	Owens DK, Davidson KW, Krist AH, Barry MJ, Cabana M, Caughey AB, Curry SJ, Doubeni CA, Epling JW, Kubik M, et al: Screening for pancreatic cancer. JAMA. 322:4382019. View Article : Google Scholar : PubMed/NCBI
2	Bray F, Ferlay J, Soerjomataram I, Siegel RL, Torre LA and Jemal A: Global cancer statistics 2018: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries. CA Cancer J Clin. 68:394–424. 2018. View Article : Google Scholar : PubMed/NCBI
3	Moffat GT, Epstein AS and O'Reilly EM: Pancreatic cancer-A disease in need: Optimizing and integrating supportive care. Cancer. 125:3927–3935. 2019. View Article : Google Scholar : PubMed/NCBI
4	Rahib L, Fleshman JM, Matrisian LM and Berlin JD: Evaluation of pancreatic cancer clinical trials and benchmarks for clinically meaningful future trials: A systematic review. JAMA Oncol. 2:1209–1216. 2016. View Article : Google Scholar : PubMed/NCBI
5	Rahib L, Smith BD, Aizenberg R, Rosenzweig AB, Fleshman JM and Matrisian LM: Projecting cancer incidence and deaths to 2030: The unexpected burden of thyroid, liver, and pancreas cancers in the United States. Cancer Res. 74:2913–2921. 2014. View Article : Google Scholar : PubMed/NCBI
6	Shi Y, Gao W, Lytle NK, Huang P, Yuan X, Dann AM, Ridinger-Saison M, Del Giorno KE, Antal CE, Liang G, et al: Targeting LIF-mediated paracrine in- teraction for pancreatic cancer therapy and monitoring. Nature. 569:131–135. 2019. View Article : Google Scholar : PubMed/NCBI
7	Fox RG, Lytle NK, Jaquish DV, Park FD, Ito T, Bajaj J, Claire S, Koechlein CS, Zimdahl B, Yano M, et al: Image based detection and targeting of therapy resistance in pancreatic adenocarcinoma. Nature. 534:407–411. 2016. View Article : Google Scholar : PubMed/NCBI
8	Karmazanovsky G, Fedorov V, Kubyshkin V and Kotchatkov A: Pancreatic head cancer: Accuracy of CT in determination of resectability. Abdom Imaging. 30:488–500. 2005. View Article : Google Scholar : PubMed/NCBI
9	Strobel O, Neoptolemos J, Jager D and Buchler MW: Optimizing the outcomes of pancreatic cancer surgery. Nat Rev Clin Oncol. 16:11–26. 2019. View Article : Google Scholar : PubMed/NCBI
10	Amrutkar M and Gladhaug I: Pancreatic cancer chemoresistance to gemcita-bine. Cancers. 9:1572017. View Article : Google Scholar
11	Feng H, Zhao X, Guo Q, Feng Y, Ma M, Guo W, Dong X, Deng C, Li C, Song X, et al: Autophagy resists EMT process to maintain retinal pigment epithelium homeostasis. Int J Biol Sci. 15:507–521. 2019. View Article : Google Scholar : PubMed/NCBI
12	Pastushenko I and Blanpain C: EMT transition states during tumor pro-gression and metastasis. Trends Cell Biol. 29:212–226. 2019. View Article : Google Scholar : PubMed/NCBI
13	Rhim AD, Mirek ET, Aiello NM, Maitra A, Bailey JM, McAllister F, Reichert M, Beatty GL, Rustgi AK, Vonderheide RH, et al: EMT and dissemination precede pancreatic tumor formation. Cell. 148:349–361. 2012. View Article : Google Scholar : PubMed/NCBI
14	De Craene B and Berx G: Regulatory networks defining EMT during cancer initiation and progression. Nat Rev Cancer. 13:97–110. 2013. View Article : Google Scholar : PubMed/NCBI
15	Costanza B, Rademaker G, Tiamiou A, De Tullio P, Leenders J, Blomme A, Bellier J, Bianchi E, Turtoi A, Delvenne P, et al: Transforming growth factor beta-induced, an extracellular matrix interacting protein, enhances glycolysis and promotes pancreatic cancer cell migration. Int J Cancer. 145:1570–1584. 2019. View Article : Google Scholar : PubMed/NCBI
16	Zhang N, Liu Y, Wang Y, Zhao M, Tu L and Luo F: Decitabine reverses TGF-β1-induced epithelial-mesenchymal transition in non-small-cell lung cancer by regulating miR-200/ZEB axis. Drug Des Devel Ther. 11:969–983. 2017. View Article : Google Scholar : PubMed/NCBI
17	Enomoto A, Murakami H, Asai N, Morone N, Watanabe T, Kawai K, Murakumo Y, Usukura J, Kaibuchi K and Takahashi M: Akt/PKB regulates actin organization and cell motility via Girdin/APE. Dev Cell. 9:389–402. 2005. View Article : Google Scholar : PubMed/NCBI
18	Weng L, Enomoto A, Ishida-Takagishi M, Asai N and Takahashi M: Girding for migratory cues: Roles of the Akt substrate Girdin in cancer progression and angiogenesis. Cancer Sci. 101:836–842. 2010. View Article : Google Scholar : PubMed/NCBI
19	Anai M, Shojima N, Katagiri H, Ogihara T, Sakoda H, Onishi Y, Ono H, Fujishiro M, Fukushima Y, Horike N, et al: A novel protein kinase B (PKB)/AKT-binding protein enhances PKB kinase activity and regulates DNA synthesis. J Biol Chem. 280:18525–18535. 2005. View Article : Google Scholar : PubMed/NCBI
20	Jiang P, Enomoto A, Jijiwa M, Kato T, Hasegawa T, Ishida M, Sato T, Asai N, Murakumo Y and Takahashi M: An actin-binding protein Girdin regulates the motility of breast cancer cells. Cancer Res. 68:1310–1318. 2008. View Article : Google Scholar : PubMed/NCBI
21	Wang X, Enomoto A, Weng L, Mizutani Y, Abudureyimu S, Esaki N, Tsuyuki Y, Chen C, Mii S, Asai N, et al: Girdin/GIV regulates collective cancer cell migration by controlling cell adhesion and cytoskeletal organization. Cancer Sci. 109:3643–3656. 2018. View Article : Google Scholar : PubMed/NCBI
22	Natsume A, Kato T, Kinjo S, Enomoto A, Toda H, Shimato S, Ohka F, Motomura K, Kondo Y, Miyata T, et al: Girdin maintains the stemness of glioblastoma stem cells. Oncogene. 31:2715–2724. 2012. View Article : Google Scholar : PubMed/NCBI
23	Yamamura Y, Asai N, Enomoto A, Kato T, Mii S, Kondo Y, Ushida K, Niimi K, Tsunoda N, Nagino M, et al: Akt-Girdin signaling in cancer-associated fibroblasts contributes to tumor progression. Cancer Res. 75:813–823. 2015. View Article : Google Scholar : PubMed/NCBI
24	Choi J, Kim KH, Oh E, Shin YK, Seo J, Kim S, Park S and Choi Y: Girdin protein expression is associated with poor prognosis in patients with invasive breast cancer. Pathology. 49:618–626. 2017. View Article : Google Scholar : PubMed/NCBI
25	Ghosh P, Tie J, Muranyi A, Singh S, Brunhoeber P, Leith K, Bowermaster R, Liao Z, Zhu Y, LaFleur B, et al: Girdin (GIV) expression as a prognostic marker of recurrence in mismatch repair-proficient stage ii colon cancer. Clin Cancer Res. 22:3488–3498. 2016. View Article : Google Scholar : PubMed/NCBI
26	Shibata T, Matsuo Y, Shamoto T, Hirokawa T, Tsuboi K, Takahashi H, Ishiguro H, Kimura M, Takeyama H and Inagaki H: Girdin, a regulator of cell motility, is a potential prognostic marker for esophageal squamous cell carcinoma. Oncol Rep. 29:2127–2132. 2013. View Article : Google Scholar : PubMed/NCBI
27	Gu F, Wang L, He J, Liu X, Zhang H, Li W, Fu L and Ma Y: Girdin, an actin-binding protein, is critical for migration, adhesion, and invasion of human glioblastoma cells. J Neurochem. 131:457–469. 2014. View Article : Google Scholar : PubMed/NCBI
28	Weng L, Han Y, Enomoto A, Kitaura Y, Nagamori S, Kanai Y, Asai N, An J, Takagishi M, Asai M, et al: Negative regulation of amino acid signaling by MAPK-regulated 4F2hc/Girdin complex. PLoS Biol. 16:e20050902018. View Article : Google Scholar : PubMed/NCBI
29	Garcia-Marcos M, Ear J, Farquhar MG and Ghosh P: A GDI (AGS3) and a GEF (GIV) regulate autophagy by balancing G protein activity and growth factor signals. Mol Biol Cell. 22:673–686. 2011. View Article : Google Scholar : PubMed/NCBI
30	Wang S, Lei Y, Cai Z, Ye X, Li L, Luo X and Yu C: Girdin regulates the proliferation and apoptosis of pancreatic cancer cells via the PI3K/Akt signalling pathway. Oncol Rep. 40:599–608. 2018.PubMed/NCBI
31	Livak JK and Schmittgen TD: Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) method. Methods. 25:402–408. 2001. View Article : Google Scholar : PubMed/NCBI
32	Wang C, Qiao C, Wang R and Zhou W: KiSS-1-mediated suppression of the invasive ability of human pancreatic carcinoma cells is not dependent on the level of KiSS-1 receptor GPR54. Mol Med Rep. 13:123–129. 2016. View Article : Google Scholar : PubMed/NCBI
33	Si W, Liu X, Wei R, Zhang Y, Zhao Y, Cui L and Hong T: MTA2-mediated inhibition of PTEN leads to pancreatic ductal adenocarcinoma carcinogenicity. Cell Death Dis. 10:2062019. View Article : Google Scholar : PubMed/NCBI
34	Huang J, Mei H, Tang Z, Li J, Zhang X, Lu Y, Huang F, Jin Q and Wang Z: Triple-amiRNA VEGFRs inhibition in pancreatic cancer improves the effificacy of chemotherapy through EMT regulation. J Cont Rele. 245:1–14. 2017. View Article : Google Scholar
35	Aiello NM, Brabletz T, Kang Y, Nieto MA, Weinberg RA and Stanger BZ: Upholding a role for EMT in pancreatic cancer metastasis. Nature. 547:E7–E8. 2017. View Article : Google Scholar : PubMed/NCBI
36	Braitsch CM, Azizoglu DB, Htike Y, Barlow HR, Schnell U, Chaney CP, Carroll TJ, Stanger BZ and Cleaver O: LATS1/2 suppress NFκB and aberrant EMT initiation to permit pancreatic progenitor differentiation. PLoS Biol. 17:e30003822019. View Article : Google Scholar : PubMed/NCBI
37	Lekka K, Tzitzi E, Giakoustidis A, Papadopoulos V and Giakoustidis D: Contemporary management of borderline resectable pancreatic ductal adenocarcinoma. Ann Hepatobiliary Pancreat Surg. 23:972019. View Article : Google Scholar : PubMed/NCBI
38	Bu JQ and Chen F: TGF-β1 promotes cells invasion and migration by inducing epithelial mesenchymal transformation in oral squamous cell carcinoma. Eur Rev Med Pharmacol Sci. 21:2137–2144. 2017.PubMed/NCBI
39	Zhang Y, Li JH, Yuan QG, Cao G and Yang WB: Upregulation of LASP2 inhibits pancreatic cancer cell migration and invasion through suppressing TGF-β-induced EMT. J Cell Biochem. 120:13651–13657. 2019. View Article : Google Scholar : PubMed/NCBI
40	Mann KM, Ying H, Juan J, Jenkins NA and Copeland NG: KRAS-related proteins in pancreatic cancer. Pharmacol Ther. 168:29–42. 2016. View Article : Google Scholar : PubMed/NCBI
41	Song M, Bode AM, Dong Z and Lee MH: AKT as a therapeutic target for cancer. Cancer Res. 79:1019–1031. 2019. View Article : Google Scholar : PubMed/NCBI
42	Martini M, De Santis MC, Braccini L, Gulluni F and Hirsch E: PI3K/AKT signaling pathway and cancer: An updated review. Ann Med. 46:372–383. 2014. View Article : Google Scholar : PubMed/NCBI
43	Yamamoto S, Tomita Y, Hoshida Y, Morooka T, Nagano H, Dono K, Umeshita K, Sakon M, Ishikawa O, Ohigashi H, et al: Prognostic significance of activated Akt expression in pancreatic ductal adenocarcinoma. Clin Cancer Res. 10:2846–2850. 2004. View Article : Google Scholar : PubMed/NCBI
44	Stoll V, Calleja V, Vassaux G, Downward J and Lemoine NR: Dominant negative inhibitors of signalling through the phosphoinositol 3-kinase pathway for gene therapy of pancreatic cancer. Gut. 54:109–116. 2005. View Article : Google Scholar : PubMed/NCBI
45	Ng SSW, Tsao MS, Chow S and Hedley DW: Inhibition of phosphatidylinositide 3-kinase enhances gemcitabine-induced apoptosis in human pancreatic cancer cells. Cancer Res. 60:5451–5455. 2000.PubMed/NCBI
46	Ni W, Fang Y, Tong L, Tong Z, Yi F, Qiu J, Wang R and Tong X: Girdin regulates the migration and invasion of glioma cells via the PI3K-Akt signaling pathway. Mol Med Rep. 12:5086–5092. 2015. View Article : Google Scholar : PubMed/NCBI
47	Mittal Y, Pavlova Y, Garcia-Marcos M and Ghosh P: Src homology domain 2-containing protein-tyrosine phosphatase-1 (SHP-1) binds and dephosphorylates G(alpha)-interacting, vesicle-associated protein (GIV)/Girdin and attenuates the GIV-phosphatidylinositol 3-kinase (PI3K)-Akt signaling pathway. J Biol Chem. 286:32404–32415. 2011. View Article : Google Scholar : PubMed/NCBI