Skip to main content
Key Specialties
Cardiology Diabetology Endocrinology Gastroenterology Geriatrics and Gerontology Gynecology and Obstetrics Hematology Infectious Disease Internal Medicine Nephrology Neurology Oncology Pediatrics Respiratory Medicine Rheumatology Surgery
Congress Coverage
2024 ACC Congress 2023 ESMO Congress 2023 EASD Congress 2023 ERS Congress 2023 ESC Congress
Clinical Collections
Advances in Alzheimer’s

At a glance: The STEP trials

A round-up of the STEP phase 3 clinical trials evaluating semaglutide for weight loss in people with overweight or obesity.
ADVANCED SEARCH
Log in
Top
BMC Cancer
Published in: BMC Cancer 1/2014

Open Access 01-12-2014 | Research article

Increased diacylglycerol kinase ζ expression in human metastatic colon cancer cells augments Rho GTPase activity and contributes to enhanced invasion

Authors: Kun Cai, Kirk Mulatz, Ryan Ard, Thanh Nguyen, Stephen H Gee

Published in: BMC Cancer | Issue 1/2014

Login to get access

Abstract

Background

Unraveling the signaling pathways responsible for the establishment of a metastatic phenotype in carcinoma cells is critically important for understanding the pathology of cancer. The acquisition of cell motility is a key property of metastatic tumor cells and is a prerequisite for invasion. Rho GTPases regulate actin cytoskeleton reorganization and the cellular responses required for cell motility and invasion. Diacylglycerol kinase ζ (DGKζ), an enzyme that phosphorylates diacylglycerol to yield phosphatidic acid, regulates the activity of the Rho GTPases Rac1 and RhoA. DGKζ mRNA is highly expressed in several different colon cancer cell lines, as well as in colon cancer tissue relative to normal colonic epithelium, and thus may contribute to the metastatic process.

Methods

To investigate potential roles of DGKζ in cancer metastasis, a cellular, isogenic model of human colorectal cancer metastatic transition was used. DGKζ protein levels, Rac1 and RhoA activity, and PAK phosphorylation were measured in the non-metastatic SW480 adenocarcinoma cell line and its highly metastatic variant, the SW620 line. The effect of DGKζ silencing on Rho GTPase activity and invasion through Matrigel-coated Transwell inserts was studied in SW620 cells. Invasiveness was also measured in PC-3 prostate cancer and MDA-MB-231 breast cancer cells depleted of DGKζ.

Results

DGKζ protein levels were elevated approximately 3-fold in SW620 cells compared to SW480 cells. There was a concomitant increase in active Rac1 in SW620 cells, as well as substantial increases in the expression and phosphorylation of the Rac1 effector PAK1. Similarly, RhoA activity and expression were increased in SW620 cells. Knockdown of DGKζ expression in SW620 cells by shRNA-mediated silencing significantly reduced Rac1 and RhoA activity and attenuated the invasiveness of SW620 cells in vitro. DGKζ silencing in highly metastatic MDA-MB-231 breast cancer cells and PC-3 prostate cancer cells also significantly attenuated their invasiveness.

Conclusion

Elevated DGKζ expression contributes to increased Rho GTPase activation and the enhanced motility of metastatic cancer cells. These findings warrant further investigation of the clinical relevance of DGKζ upregulation in colon and other cancers. Interfering with DGKζ function could provide a means of inhibiting invasion and metastasis.
Appendix
Available only for authorised users
Literature
1.
Siegel R, Naishadham D, Jemal A: Cancer statistics, 2012. CA Cancer J Clin. 2012, 62: 10-29. 10.3322/caac.20138.CrossRefPubMed
2.
Fearon ER, Vogelstein B: A genetic model for colorectal tumorigenesis. Cell. 1990, 61: 759-767. 10.1016/0092-8674(90)90186-I.CrossRefPubMed
3.
Kinzler KW, Vogelstein B: Lessons from hereditary colorectal cancer. Cell. 1996, 87: 159-170. 10.1016/S0092-8674(00)81333-1.CrossRefPubMed
4.
Vogelstein B, Fearon ER, Hamilton SR, Kern SE, Preisinger AC, Leppert M, Nakamura Y, White R, Smits AM, Bos JL: Genetic alterations during colorectal-tumor development. N Engl J Med. 1988, 319: 525-532. 10.1056/NEJM198809013190901.CrossRefPubMed
5.
Provenzani A, Fronza R, Loreni F, Pascale A, Amadio M, Quattrone A: Global alterations in mRNA polysomal recruitment in a cell model of colorectal cancer progression to metastasis. Carcinogenesis. 2006, 27: 1323-1333. 10.1093/carcin/bgi377.CrossRefPubMed
6.
Hanahan D, Weinberg RA: The hallmarks of cancer. Cell. 2000, 100: 57-70. 10.1016/S0092-8674(00)81683-9.CrossRefPubMed
7.
Thiery JP, Sleeman JP: Complex networks orchestrate epithelial-mesenchymal transitions. Nat Rev Mol Cell Biol. 2006, 7: 131-142. 10.1038/nrm1835.CrossRefPubMed
8.
Yilmaz M, Christofori G: EMT, the cytoskeleton, and cancer cell invasion. Cancer Metastasis Rev. 2009, 28: 15-33. 10.1007/s10555-008-9169-0.CrossRefPubMed
9.
Hall A: Rho GTPases and the Actin Cytoskeleton. Science. 1998, 279: 509-514. 10.1126/science.279.5350.509.CrossRefPubMed
10.
Ridley AJ, Paterson HF, Johnston CL, Diekmann D, Hall A: The small GTP-binding protein rac regulates growth factor-induced membrane ruffling. Cell. 1992, 70: 401-410. 10.1016/0092-8674(92)90164-8.CrossRefPubMed
11.
Hall A: The cytoskeleton and cancer. Cancer Metastasis Rev. 2009, 28: 5-14. 10.1007/s10555-008-9166-3.CrossRefPubMed
12.
Raftopoulou M, Hall A: Cell migration: Rho GTPases lead the way. Dev Biol. 2004, 265: 23-32. 10.1016/j.ydbio.2003.06.003.CrossRefPubMed
13.
Gomez del Pulgar T, Benitah SA, Valeron PF, Espina C, Lacal JC: Rho GTPase expression in tumourigenesis: evidence for a significant link. Bioessays. 2005, 27: 602-613. 10.1002/bies.20238.CrossRefPubMed
14.
Clark EA, Golub TR, Lander ES, Hynes RO: Genomic analysis of metastasis reveals an essential role for RhoC. Nature. 2000, 406: 532-535. 10.1038/35020106.CrossRefPubMed
15.
Keely PJ, Westwick JK, Whitehead IP, Der CJ, Parise LV: Cdc42 and Rac1 induce integrin-mediated cell motility and invasiveness through PI(3)K. Nature. 1997, 390: 632-636. 10.1038/37656.CrossRefPubMed
16.
Knight-Krajewski S, Welsh CF, Liu Y, Lyons LS, Faysal JM, Yang ES, Burnstein KL: Deregulation of the Rho GTPase, Rac1, suppresses cyclin-dependent kinase inhibitor p21(CIP1) levels in androgen-independent human prostate cancer cells. Oncogene. 2004, 23: 5513-5522. 10.1038/sj.onc.1207708.CrossRefPubMed
17.
Kurisu S, Suetsugu S, Yamazaki D, Yamaguchi H, Takenawa T: Rac-WAVE2 signaling is involved in the invasive and metastatic phenotypes of murine melanoma cells. Oncogene. 2005, 24: 1309-1319. 10.1038/sj.onc.1208177.CrossRefPubMed
18.
Michiels F, Habets GG, Stam JC, van der Kammen RA, Collard JG: A role for Rac in Tiam1-induced membrane ruffling and invasion. Nature. 1995, 375: 338-340. 10.1038/375338a0.CrossRefPubMed
19.
Fritz G, Just I, Kaina B: Rho GTPases are over-expressed in human tumors. Int J Cancer. 1999, 81: 682-687. 10.1002/(SICI)1097-0215(19990531)81:5<682::AID-IJC2>3.0.CO;2-B.CrossRefPubMed
20.
Suwa H, Ohshio G, Imamura T, Watanabe G, Arii S, Imamura M, Narumiya S, Hiai H, Fukumoto M: Overexpression of the rhoC gene correlates with progression of ductal adenocarcinoma of the pancreas. Br J Cancer. 1998, 77: 147-152. 10.1038/bjc.1998.23.CrossRefPubMedPubMedCentral
21.
Fearnhead NS, Britton MP, Bodmer WF: The ABC of APC. Hum Mol Genet. 2001, 10: 721-733. 10.1093/hmg/10.7.721.CrossRefPubMed
22.
Kawasaki Y, Sato R, Akiyama T: Mutated APC and Asef are involved in the migration of colorectal tumour cells. Nat Cell Biol. 2003, 5: 211-215. 10.1038/ncb937.CrossRefPubMed
23.
Lazer G, Katzav S: Guanine nucleotide exchange factors for RhoGTPases: good therapeutic targets for cancer therapy?. Cell Signal. 2011, 23: 969-979. 10.1016/j.cellsig.2010.10.022.CrossRefPubMed
24.
Jaffe AB, Hall A: RHO GTPASES: Biochemistry and Biology. Annu Rev Cell Dev Biol. 2005, 21: 247-269. 10.1146/annurev.cellbio.21.020604.150721.CrossRefPubMed
25.
Dermardirossian C, Bokoch GM: GDIs: central regulatory molecules in Rho GTPase activation. Trends Cell Biol. 2005, 15: 356-363. 10.1016/j.tcb.2005.05.001.CrossRefPubMed
26.
27.
Topham MK, Epand RM: Mammalian diacylglycerol kinases: molecular interactions and biological functions of selected isoforms. Biochim Biophys Acta. 2009, 1790: 416-424. 10.1016/j.bbagen.2009.01.010.CrossRefPubMedPubMedCentral
28.
Abramovici H, Mojtabaie P, Parks RJ, Zhong XP, Koretzky GA, Topham MK, Gee SH: Diacylglycerol kinase zeta regulates actin cytoskeleton reorganization through dissociation of Rac1 from RhoGDI. Mol Biol Cell. 2009, 20: 2049-2059. 10.1091/mbc.E07-12-1248.CrossRefPubMedPubMedCentral
29.
Ard R, Mulatz K, Abramovici H, Maillet JC, Fottinger A, Foley T, Byham MR, Iqbal TA, Yoneda A, Couchman JR, Parks RJ, Gee SH: Diacylglycerol kinase zeta regulates RhoA activation via a kinase-independent scaffolding mechanism. Mol Biol Cell. 2012, 23: 4008-4019. 10.1091/mbc.E12-01-0026.CrossRefPubMedPubMedCentral
30.
Sabates-Bellver J, Van der Flier LG, de Palo M, Cattaneo E, Maake C, Rehrauer H, Laczko E, Kurowski MA, Bujnicki JM, Menigatti M, Luz J, Ranalli TV, Gomes V, Pastorelli A, Faggiani R, Anti M, Jiricny J, Clevers H, Marra G: Transcriptome profile of human colorectal adenomas. Mol Cancer Res. 2007, 5: 1263-1275. 10.1158/1541-7786.MCR-07-0267.CrossRefPubMed
31.
Leibovitz A, Stinson JC, McCombs WB, McCoy CE, Mazur KC, Mabry ND: Classification of human colorectal adenocarcinoma cell lines. Cancer Res. 1976, 36: 4562-4569.PubMed
32.
Abramovici H, Hogan AB, Obagi C, Topham MK, Gee SH: Diacylglycerol kinase-zeta localization in skeletal muscle is regulated by phosphorylation and interaction with syntrophins. Mol Biol Cell. 2003, 14: 4499-4511. 10.1091/mbc.E03-03-0190.CrossRefPubMedPubMedCentral
33.
Topham MK, Bunting M, Zimmerman GA, McIntyre TM, Blackshear PJ, Prescott SM: Protein kinase C regulates the nuclear localization of diacylglycerol kinase-zeta. Nature. 1998, 394: 697-700. 10.1038/29337.CrossRefPubMed
34.
Yakubchyk Y, Abramovici H, Maillet JC, Daher E, Obagi C, Parks RJ, Topham MK, Gee SH: Regulation of neurite outgrowth in N1E-115 cells through PDZ-mediated recruitment of diacylglycerol kinase zeta. Mol Cell Biol. 2005, 25: 7289-7302. 10.1128/MCB.25.16.7289-7302.2005.CrossRefPubMedPubMedCentral
35.
Sells MA, Pfaff A, Chernoff J: Temporal and spatial distribution of activated Pak1 in fibroblasts. J Cell Biol. 2000, 151: 1449-1458. 10.1083/jcb.151.7.1449.CrossRefPubMedPubMedCentral
36.
Sander EE, van Delft S, ten Klooster JP, Reid T, van der Kammen RA, Michiels F, Collard JG: Matrix-dependent Tiam1/Rac signaling in epithelial cells promotes either cell-cell adhesion or cell migration and is regulated by phosphatidylinositol 3-kinase. J Cell Biol. 1998, 143: 1385-1398. 10.1083/jcb.143.5.1385.CrossRefPubMedPubMedCentral
37.
Bokoch GM: Biology of the p21-activated kinases. Annu Rev Biochem. 2003, 72: 743-781. 10.1146/annurev.biochem.72.121801.161742.CrossRefPubMed
38.
Kim HR, Wheeler MA, Wilson CM, Iida J, Eng D, Simpson MA, McCarthy JB, Bullard KM: Hyaluronan facilitates invasion of colon carcinoma cells in vitro via interaction with CD44. Can Res. 2004, 64: 4569-4576. 10.1158/0008-5472.CAN-04-0202.CrossRef
39.
40.
Zhu XL, Liang L, Ding YQ: Overexpression of FMNL2 is closely related to metastasis of colorectal cancer. Int J Colorectal Dis. 2008, 23: 1041-1047. 10.1007/s00384-008-0520-2.CrossRefPubMed
41.
Chianale F, Cutrupi S, Rainero E, Baldanzi G, Porporato PE, Traini S, Filigheddu N, Gnocchi VF, Santoro MM, Parolini O, van Blitterswijk WJ, Sinigaglia F, Graziani A: Diacylglycerol kinase-alpha mediates hepatocyte growth factor-induced epithelial cell scatter by regulating Rac activation and membrane ruffling. Mol Biol Cell. 2007, 18: 4859-4871. 10.1091/mbc.E07-02-0177.CrossRefPubMedPubMedCentral
42.
Chianale F, Rainero E, Cianflone C, Bettio V, Pighini A, Porporato PE, Filigheddu N, Serini G, Sinigaglia F, Baldanzi G, Graziani A: Diacylglycerol kinase alpha mediates HGF-induced Rac activation and membrane ruffling by regulating atypical PKC and RhoGDI. Proc Natl Acad Sci USA. 2010, 107: 4182-4187. 10.1073/pnas.0908326107.CrossRefPubMedPubMedCentral
43.
Tsushima S, Kai M, Yamada K, Imai S, Houkin K, Kanoh H, Sakane F: Diacylglycerol kinase gamma serves as an upstream suppressor of Rac1 and lamellipodium formation. J Biol Chem. 2004, 279: 28603-28613. 10.1074/jbc.M314031200.CrossRefPubMed
44.
Friedl P, Wolf K: Tumour-cell invasion and migration: diversity and escape mechanisms. Nat Rev Cancer. 2003, 3: 362-374. 10.1038/nrc1075.CrossRefPubMed
45.
Kamai T, Tsujii T, Arai K, Takagi K, Asami H, Ito Y, Oshima H: Significant association of Rho/ROCK pathway with invasion and metastasis of bladder cancer. Clin Cancer Res. 2003, 9: 2632-2641.PubMed
46.
Takami Y, Higashi M, Kumagai S, Kuo PC, Kawana H, Koda K, Miyazaki M, Harigaya K: The activity of RhoA is correlated with lymph node metastasis in human colorectal cancer. Dig Dis Sci. 2008, 53: 467-473. 10.1007/s10620-007-9887-0.CrossRefPubMed
47.
van Golen KL, Davies S, Wu ZF, Wang Y, Bucana CD, Root H, Chandrasekharappa S, Strawderman M, Ethier SP, Merajver SD: A novel putative low-affinity insulin-like growth factor-binding protein, LIBC (lost in inflammatory breast cancer), and RhoC GTPase correlate with the inflammatory breast cancer phenotype. Clin Cancer Res. 1999, 5: 2511-2519.PubMed
48.
Wang L, Yang L, Luo Y, Zheng Y: A Novel Strategy for Specifically Down-regulating Individual Rho GTPase Activity in Tumor Cells. J Biol Chem. 2003, 278: 44617-44625. 10.1074/jbc.M308929200.CrossRefPubMed
49.
Espina C, Cespedes MV, Garcia-Cabezas MA, Gomez del Pulgar MT, Boluda A, Oroz LG, Benitah SA, Cejas P, Nistal M, Mangues R, Lacal JC: A critical role for Rac1 in tumor progression of human colorectal adenocarcinoma cells. Am J Pathol. 2008, 172: 156-166. 10.2353/ajpath.2008.070561.CrossRefPubMedPubMedCentral
50.
Walker K, Olson MF: Targeting Ras and Rho GTPases as opportunities for cancer therapeutics. Curr Opin Genet Dev. 2005, 15: 62-68. 10.1016/j.gde.2004.11.001.CrossRefPubMed
51.
Shankavaram UT, Reinhold WC, Nishizuka S, Major S, Morita D, Chary KK, Reimers MA, Scherf U, Kahn A, Dolginow D, Cossman J, Kaldjian EP, Scudiero DA, Petricoin E, Liotta L, Lee JK, Weinstein JN: Transcript and protein expression profiles of the NCI-60 cancer cell panel: an integromic microarray study. Mol Cancer Ther. 2007, 6: 820-832. 10.1158/1535-7163.MCT-06-0650.CrossRefPubMed
52.
Su AI, Welsh JB, Sapinoso LM, Kern SG, Dimitrov P, Lapp H, Schultz PG, Powell SM, Moskaluk CA, Frierson HF, Hampton GM: Molecular classification of human carcinomas by use of gene expression signatures. Can Res. 2001, 61: 7388-7393.
53.
Barretina J, Caponigro G, Stransky N, Venkatesan K, Margolin AA, Kim S, Wilson CJ, Lehar J, Kryukov GV, Sonkin D, Reddy A, Liu M, Murray L, Berger MF, Monahan JE, Morais P, Meltzer J, Korejwa A, Jane-Valbuena J, Mapa FA, Thibault J, Bric-Furlong E, Raman P, Shipway A, Engels IH, Cheng J, Yu GK, Yu J, Aspesi P, de Silva M, et al: The Cancer Cell Line Encyclopedia enables predictive modelling of anticancer drug sensitivity. Nature. 2012, 483: 603-607. 10.1038/nature11003.CrossRefPubMedPubMedCentral
Metadata
Title
Increased diacylglycerol kinase ζ expression in human metastatic colon cancer cells augments Rho GTPase activity and contributes to enhanced invasion
Authors
Kun Cai
Kirk Mulatz
Ryan Ard
Thanh Nguyen
Stephen H Gee
Publication date
01-12-2014
Publisher
BioMed Central
Published in
BMC Cancer / Issue 1/2014
Electronic ISSN: 1471-2407
DOI
https://doi.org/10.1186/1471-2407-14-208

Other articles of this Issue 1/2014

BMC Cancer 1/2014 Go to the issue

Case report

Solitary breast metastasis from myxoid liposarcoma

Research article

Clearing the air: improving smoke-free policy compliance at the national oncology hospital in Armenia

Research article

Integrated genomic study of quadruple-WT GIST (KIT/PDGFRA/SDH/RAS pathway wild-type GIST)

Research article

Incidence and outcome for patients with occult lymph node involvement in T1 and T2 oral squamous cell carcinoma: a prospective study

Reviewer acknowledgement

Reviewer acknowledgement 2013

Technical advance

Comparative evaluation of the new FDA approved THxID™-BRAF test with high resolution melting and sanger sequencing
Webinar | 19-02-2024 | 17:30 (CET)

Keynote webinar | Spotlight on antibody–drug conjugates in cancer

Watch now

Antibody–drug conjugates (ADCs) are novel agents that have shown promise across multiple tumor types. Explore the current landscape of ADCs in breast and lung cancer with our experts, and gain insights into the mechanism of action, key clinical trials data, existing challenges, and future directions.

Dr. Véronique Diéras
Prof. Fabrice Barlesi
Developed by: Springer Medicine
Image Credits