Contribution of C3G and other GEFs to liver cancer development and progression

Abstract

Primary liver cancers constitute the fourth leading cause of cancer mortality worldwide, due to their high morbidity, late diagnosis and lack of effective treatments. Hepatocellular carcinoma (HCC) represents 80% and cholangiocarcinoma (CCA) 15% of liver cancers. Several genetic and epigenetic gene alterations (e.g., TERT, TP53 or CTNNB1) are HCC drivers, although many additional gene alterations contribute to HCC initiation and/or progression. Rho and Ras GTPases have been widely implicated in tumorigenesis and their activators (GEFs) have recently emerged as putative key players in liver cancer. The Ras GEF, C3G (RAPGEF1), a GEF mainly for Rap proteins, has recently been uncovered as a relevant gene in HCC. Its upregulation promotes tumor growth, although a decrease in C3G levels favors migration/invasion and lung metastasis. Rap1A/1B/2A/2B are overexpressed in HCC tumors, but their effects are controversial and not equivalent to those of C3G. The C3G partner, CRKL, is also overexpressed in HCC, promoting proliferation, migration and invasion. Various Rho GEFs are also deregulated in liver cancer. Tiam1 and Tiam2 expression is upregulated in HCC, promoting proliferation, migration and metastasis. In addition, ARHGEF-10L/9/19/39 are overexpressed in HCC tumors, facilitating migration, invasion, metastasis and proliferation. Another Rho GEF, Vav2, is also involved in metastasis. Little is known about the participation of these GEFs and GTPases in CCA. However, analysis of cancer databases uncovered deregulations or genetic alterations in several of these genes, in both CCA and HCC. Hence, GEFs function appear essential for liver homeostasis, although future studies are needed to define their precise function in liver cancer.

Keywords

Liver cancer / C3G / RAPGEF1 / Rap / Ras GEFs / Rho GEFs / CRK / hepatocarcinoma / cholangiocarcinoma

References

[1] Bray F,Soerjomataram I,Torre LA.Global cancer statistics 2018: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries.CA Cancer J Clin2018;68:394-424

[2] Craig AJ,Garcia-Lezana T,Villanueva A.Tumour evolution in hepatocellular carcinoma.Nat Rev Gastroenterol Hepatol2020;17:139-52

[3] Banales JM,Lamarca A.Cholangiocarcinoma 2020: the next horizon in mechanisms and management.Nat Rev Gastroenterol Hepatol2020;17:557-88 PMCID:PMC7447603

[4] Wang G,Liang N.Oncogenic driver genes and tumor microenvironment determine the type of liver cancer.2020;11:313 PMCID:PMC7198508

[5] Arechederra M,Yim A.Hypermethylation of gene body CpG islands predicts high dosage of functional oncogenes in liver cancer.Nat Commun2018;9:3164 PMCID:PMC6082886

[6] Colicelli J.Human RAS superfamily proteins and related GTPases.Sci STKE2004;2004:RE13 PMCID:PMC2828947

[7] Vetter IR.The guanine nucleotide-binding switch in three dimensions.Science2001;294:1299-304

[8] Cherfils J.Regulation of small GTPases by GEFs, GAPs, and GDIs.Physiol Rev2013;93:269-309

[9] Li S,Counter CM.A model for RAS mutation patterns in cancers: finding the sweet spot.Nat Rev Cancer2018;18:767-77

[10] Kazanietz MG.The Rac GTPase in Cancer: From Old Concepts to New Paradigms.Cancer Res2017;77:5445-51 PMCID:PMC5645227

[11] Casado-Medrano V,Lopez-Haber C.The role of Rac in tumor susceptibility and disease progression: from biochemistry to the clinic.Biochem Soc Trans2018;46:1003-12 PMCID:PMC6261428

[12] Bustelo XR.RHO GTPases in cancer: known facts, open questions, and therapeutic challenges.Biochem Soc Trans2018;46:741-60

[13] Sahai E.RHO-GTPases and cancer.Nat Rev Cancer2002;2:133-42

[14] Reuther GW.The Ras branch of small GTPases: Ras family members don't fall far from the tree.Curr Opin Cell Biol2000;12:157-65

[15] Bos JL,Wittinghofer A.GEFs and GAPs: critical elements in the control of small G proteins.Cell2007;129:865-77

[16] Rebhun JF,Quilliam LA.Identification and characterization of a new family of guanine nucleotide exchange factors for the ras-related GTPase Ral.J Biol Chem2000;275:13406-10

[17] Ferro E.RalGDS family members couple Ras to Ral signalling and that's not all.Cell Signal2010;22:1804-10

[18] Ridley AJ.Rho GTPases and actin dynamics in membrane protrusions and vesicle trafficking.Trends Cell Biol2006;16:522-5229

[19] Rossman KL,Sondek J.GEF means go: turning on RHO GTPases with guanine nucleotide-exchange factors.Nat Rev Mol Cell Biol2005;6:167-80

[20] Cote JF.GEF what?.Trends Cell Biol2007;17:383-93 PMCID:PMC2887429

[21] Carabias A,de Cima S.Mechanisms of autoregulation of C3G, activator of the GTPase Rap1, and its catalytic deregulation in lymphomas.Sci Signal2020;13:eabb7075

[22] Vigil D,Rossman KL.Ras superfamily GEFs and GAPs: validated and tractable targets for cancer therapy?.Nat Rev Cancer2010;10:842-57 PMCID:PMC3124093

[23] Sequera C,Guerrero C.How Rap and its GEFs control liver physiology and cancer development. C3G alterations in human hepatocarcinoma.Hepat Oncol2018;5:HEP05 PMCID:PMC6168044

[24] Spolitu S,Dai W.Hepatic Glucagon Signaling Regulates PCSK9 and Low-Density Lipoprotein Cholesterol.Circ Res2019;124:38-51 PMCID:PMC6457439

[25] Sakata H,Taylor WG.A Rho-specific exchange factor Ect2 is induced from S to M phases in regenerating mouse liver.Hepatology2000;32:193-9

[26] Bang BR,Seo GY,Kang YJ.The protein tyrosine kinase SYK regulates the alternative p38 activation in liver during acute liver inflammation.Sci Rep2019;9:17838 PMCID:PMC6882802

[27] Welch HC.Regulation and function of P-Rex family Rac-GEFs.Small GTPases2015;6:49-70 PMCID:PMC4601503

[28] Harnois T,Rioux A,Kitzis A.Differential interaction and activation of Rho family GTPases by p210bcr-abl and p190bcr-abl.Oncogene2003;22:6445-54

[29] Bodemann BO.Ral GTPases and cancer: linchpin support of the tumorigenic platform.Nat Rev Cancer2008;8:133-40

[30] Cook DR,Der CJ.Rho guanine nucleotide exchange factors: regulators of Rho GTPase activity in development and disease.Oncogene2014;33:4021-35 PMCID:PMC4875565

[31] Gotoh T,Nakamura S.Identification of Rap1 as a target for the Crk SH3 domain-binding guanine nucleotide-releasing factor C3G.Mol Cell Biol1995;15:6746-53 PMCID:PMC230928

[32] Gotoh T,Tokuda M.Activation of R-Ras by Ras-Guanine Nucleotide-releasing Factor*.J Cancer Res Clin Oncol1997;272:18602-7

[33] Guerrero C,Fernández-Medarde A.C3G-mediated suppression of oncogene-induced focus formation in fibroblasts involves inhibition of ERK activation, cyclin A expression and alterations of anchorage-independent growth.Oncogene2004;23:4885-93

[34] Gutiérrez-Uzquiza A,Molina I.C3G down-regulates p38 MAPK activity in response to stress by Rap-1 independent mechanisms: involvement in cell death.Cell Signal2010;22:533-42

[35] Shakyawar DK,Ramadhas A,Radha V.C3G shows regulated nucleocytoplasmic exchange and represses histone modifications associated with euchromatin.Mol Biol Cell2017;28:984-95 PMCID:PMC5385946

[36] Shakyawar DK,Radha V.C3G dynamically associates with nuclear speckles and regulates mRNA splicing.Mol Biol Cell2018;29:1111-24 PMCID:PMC5921577

[37] Radha V,Dayma K.Signalling to actin: role of C3G, a multitasking guanine-nucleotide-exchange factor.Biosci Rep2011;31:231-44

[38] Ohba Y,Ogura A.Requirement for C3G-dependent Rap1 activation for cell adhesion and embryogenesis.EMBO J2001;20:3333-41 PMCID:PMC125518

[39] Ortiz-Rivero S,Hernandez-Cano L.C3G, through its GEF activity, induces megakaryocytic differentiation and proplatelet formation.Cell Commun Signal2018;16:101 PMCID:PMC6299959

[40] Guerrero C,Rojas JM,Esteban LM.Transformation suppressor activity of C3G is independent of its CDC25-homology domain.Oncogene1998;16:613-24

[41] Okino K,Nakayama H.Inactivation of Crk SH3 domain-binding guanine nucleotide-releasing factor (C3G) in cervical squamous cell carcinoma.Int J Gynecol Cancer2006;16:763-71

[42] Priego N,Sequera C.C3G knock-down enhances migration and invasion by increasing Rap1-mediated p38alpha activation, while it impairs tumor growth through p38alpha-independent mechanisms.Oncotarget2016;7:45060-78 PMCID:PMC5216706

[43] Dayma K.Cytoskeletal remodeling by C3G to induce neurite-like extensions and inhibit motility in highly invasive breast carcinoma cells.Biochim Biophys Acta2011;1813:456-65

[44] Hirata T,Koizumi K.Amplification, up-regulation and over-expression of C3G (CRK SH3 domain-binding guanine nucleotide-releasing factor) in non-small cell lung cancers.J Hum Genet2004;49:290-5

[45] Gutierrez-Berzal J,Martin-Encabo S.Characterization of p87C3G, a novel, truncated C3G isoform that is overexpressed in chronic myeloid leukemia and interacts with Bcr-Abl.Exp Cell Res2006;312:938-48

[46] Sequera C,Manzano S.C3G Is Upregulated in Hepatocarcinoma, Contributing to Tumor Growth and Progression and to HGF/MET Pathway Activation.Cancers (Basel)2020;12:2282 PMCID:PMC7463771

[47] Lin Y,Chou C.Rap1-suppressed tumorigenesis is concomitant with the interference in ras effector signaling.FEBS Lett2000;467:184-8

[48] Sheng Y,Chen K.Functional analysis of miR-101-3p and Rap1b involved in hepatitis B virus-related hepatocellular carcinoma pathogenesis.Biochem Cell Biol2014;92:152-62

[49] Zhang L,Yang YS.Knockdown of Rap2B Inhibits the Proliferation and Invasion in Hepatocellular Carcinoma Cells.Oncol Res2017;25:19-27 PMCID:PMC7840814

[50] Tang Z,Chen J.Rap1b enhances the invasion and migration of hepatocellular carcinoma cells by up-regulating Twist 1.Exp Cell Res2018;367:56-64

[51] Schippers M,Post E.Upregulation of Epac-1 in Hepatic Stellate Cells by Prostaglandin E(2) in Liver Fibrosis Is Associated with Reduced Fibrogenesis.J Pharmacol Exp Ther2017;363:126-35

[52] Zhang X,Han F.Sp1-regulated transcription of RasGRP1 promotes hepatocellular carcinoma (HCC) proliferation.Liver Int2018;38:2006-17

[53] Wang XK,Yang CK.Oncogene PLCE1 may be a diagnostic biomarker and prognostic biomarker by influencing cell cycle, proliferation, migration, and invasion ability in hepatocellular carcinoma cell lines.J Cell Physiol2020;235:7003-17

[54] Kang X,Qi X.Screening and identification of key genes between liver hepatocellular carcinoma (LIHC) and cholangiocarcinoma (CHOL) by bioinformatic analysis.Medicine (Baltimore)2020;99:e23563 PMCID:PMC7738106

[55] Shirasaki T,Shimakami T.MicroRNA-27a regulates lipid metabolism and inhibits hepatitis C virus replication in human hepatoma cells.J Virol2013;87:5270-86 PMCID:PMC3624309

[56] Calvisi DF,Conner EA.Inactivation of Ras GTPase-activating proteins promotes unrestrained activity of wild-type Ras in human liver cancer.J Hepatol2011;54:311-9 PMCID:PMC3031080

[57] Birge RB,Inagaki F.Crk and CrkL adaptor proteins: networks for physiological and pathological signaling.Cell Communication and Signaling2009;7:13 PMCID:PMC2689226

[58] Rodrigues SP,Chan G.CrkI and CrkII function as key signaling integrators for migration and invasion of cancer cells.Mol Cancer Res2005;3:183-94

[59] Guo C,Sun MZ.The role of CT10 regulation of kinase-like in cancer.Future Oncol2014;10:2687-97

[60] Srinivasan S.Increased Soluble CrkL in Serum of Breast Cancer Patients Is Associated with Advanced Disease.Cancers (Basel)2019;11 PMCID:PMC6679112

[61] Liu CH,Chau GY.Analysis of protein-protein interactions in cross-talk pathways reveals CRKL protein as a novel prognostic marker in hepatocellular carcinoma.Mol Cell Proteomics2013;12:1335-49 PMCID:PMC3650343

[62] Guo C,Zhang Q,Sun M-Z.miR-429 suppresses tumor migration and invasion by targeting CRKL in hepatocellular carcinoma via inhibiting Raf/MEK/ERK pathway and epithelial-mesenchymal transition.Scientific Reports2018;8:2375 PMCID:PMC5799248

[63] Ren Y,Li J.The long noncoding RNA PCAT-1 links the microRNA miR-215 to oncogene CRKL-mediated signaling in hepatocellular carcinoma.J Biol Chem2017;292:17939-49 PMCID:PMC5663891

[64] Guo C,Zhao D.A novel ETV6-miR-429-CRKL regulatory circuitry contributes to aggressiveness of hepatocellular carcinoma.J Exp Clin Cancer Res2020;39:70 PMCID:PMC7178969

[65] Abdul S,Wang J,Sun MZ.Bidirectional interaction of lncRNA AFAP1-AS1 and CRKL accelerates the proliferative and metastatic abilities of hepatocarcinoma cells.J Adv Res2020;24:121-30 PMCID:PMC7139140

[66] Lin Q,Guo C,Chen X.CRKL overexpression suppresses in vitro proliferation, invasion and migration of murine hepatocarcinoma Hca-P cells.Biomed Pharmacother2015;69:11-7

[67] Zhou Z,Guo C,Liu S.CRKII overexpression promotes the in vitro proliferation, migration and invasion potential of murine hepatocarcinoma Hca-P cells.Oncology letters2019;17:5169-74 PMCID:PMC6507350

[68] Sun X,Wang J.Annexin A5 regulates hepatocarcinoma malignancy via CRKI/II-DOCK180-RAC1 integrin and MEK-ERK pathways.2018;9:637 PMCID:PMC5970249

[69] Hodge RG.Regulating Rho GTPases and their regulators.Nature Reviews Molecular Cell Biology2016;17:496-510

[70] Otsuki Y,Yoshii S,Nakaya K.Tumor metastasis suppressor nm23H1 regulates Rac1 GTPase by interaction with Tiam1.Proc Natl Acad Sci U S A2001;98:4385-90 PMCID:PMC31844

[71] Malliri A.Role of Rho-family proteins in cell adhesion and cancer.Curr Opin Cell Biol2003;15:583-9

[72] Malliri A,Clark K,Michiels F.Mice deficient in the Rac activator Tiam1 are resistant to Ras-induced skin tumours.Nature2002;417:867-71

[73] Strumane K,van der Valk M.Tiam1-deficiency impairs mammary tumor formation in MMTV-c-neu but not in MMTV-c-myc mice.J Cancer Res Clin Oncol2009;135:69-80

[74] Izumi D,Ureta E,Baba H.TIAM1 promotes chemoresistance and tumor invasiveness in colorectal cancer.2019;10:267 PMCID:PMC6425043

[75] Ding Y,Wang S.Overexpression of Tiam1 in hepatocellular carcinomas predicts poor prognosis of HCC patients.Int J Cancer2009;124:653-8

[76] Yang W,Liu X,Yang W.Up-regulation of Tiam1 and Rac1 correlates with poor prognosis in hepatocellular carcinoma.Jpn J Clin Oncol2010;40:1053-9

[77] Huang J,Guan J.Tiam1 is associated with hepatocellular carcinoma metastasis.Int J Cancer2013;132:90-100

[78] Wang W,Huang DB.Tumor-associated mesenchymal stem cells promote hepatocellular carcinoma metastasis via a DNM3OS/KDM6B/TIAM1 axis.Cancer Lett2021;503:19-31

[79] Zhang Y,Li Q.Histone methyltransferase SETDB1 promotes cells proliferation and migration by interacting withTiam1 in hepatocellular carcinoma.BMC Cancer2018;18:539 PMCID:PMC5941371

[80] Chen G,Liu C,Yuan D.MicroRNA-377 suppresses cell proliferation and invasion by inhibiting TIAM1 expression in hepatocellular carcinoma.PLoS One2015;10:e0117714 PMCID:PMC4349803

[81] Liu Y,Huang J.MiR-141 suppresses the migration and invasion of HCC cells by targeting Tiam1.PLoS One2014;9:e88393 PMCID:PMC3923786

[82] Cheng W,Zuo Z.Biological effects of RNAi targeted inhibiting Tiam1 gene expression on cholangiocarcinoma cells.Int J Clin Exp Pathol2015;8:15511-26 PMCID:PMC4730034

[83] Chen JS,Leu YW,Sun HS.Expression of T-cell lymphoma invasion and metastasis 2 (TIAM2) promotes proliferation and invasion of liver cancer.Int J Cancer2012;130:1302-13

[84] Yen WH,Hung JJ,Chen JS.Sp1-mediated ectopic expression of T-cell lymphoma invasion and metastasis 2 in hepatocellular carcinoma.Cancer Med2016;5:465-77 PMCID:PMC4799941

[85] Tang J,Xu B,Ma Z.ARHGEF10L contributes to liver tumorigenesis through RhoA-ROCK1 signaling and the epithelial-mesenchymal transition.Exp Cell Res2019;374:46-68

[86] Schmidt A.Guanine nucleotide exchange factors for Rho GTPases: turning on the switch.Genes Dev2002;16:1587-609

[87] Gao J.Expression of Rho Guanine Nucleotide Exchange Factor 39 (ARHGEF39) and Its Prognostic Significance in Hepatocellular Carcinoma.Med Sci Monit2019;25:7826-35 PMCID:PMC6820342

[88] Wang H,Wang Y,Cai B.C9orf100, a new member of the Dbl-family guanine nucleotide exchange factors, promotes cell proliferation and migration in hepatocellular carcinoma.Mol Med Rep2012;5:1169-74

[89] Zhou J,Peng C,Wang W.miR-503 regulates metastatic function through Rho guanine nucleotide exchanger factor 19 in hepatocellular carcinoma.J Surg Res2014;188:129-36

[90] Chen L,Yuan YF.CHD1L promotes hepatocellular carcinoma progression and metastasis in mice and is associated with these processes in human patients.J Clin Invest2010;120:1178-91 PMCID:PMC2846051

[91] Chen J,Zhang X.ECT2 regulates the Rho/ERK signalling axis to promote early recurrence in human hepatocellular carcinoma.J Hepatol2015;62:1287-95

[92] Fang ZQ,Zhang YQ.MiR-490-5p inhibits the metastasis of hepatocellular carcinoma by down-regulating E2F2 and ECT2.J Cell Biochem2018;119:8317-24

[93] Yu Y,Wu P.MiR-490-5p inhibits the stemness of hepatocellular carcinoma cells by targeting ECT2.J Cell Biochem2019;120:967-76

[94] Gao J,Yu X,Zhou F.Upregulated microRNA-194 impairs stemness of cholangiocarcinoma cells through the Rho pathway via inhibition of ECT2.J Cell Biochem2020;121:4239-50

[95] Toksoz D.Novel human oncogene lbc detected by transfection with distinct homology regions to signal transduction products.Oncogene1994;9:621-8

[96] Sterpetti P,Candelaresi C.Cell proliferation and drug resistance in hepatocellular carcinoma are modulated by Rho GTPase signals.Am J Physiol Gastrointest Liver Physiol2006;290:G624-32

[97] Ahronian LG,Chen YW,Klimstra DS.A novel KLF6-Rho GTPase axis regulates hepatocellular carcinoma cell migration and dissemination.Oncogene2016;35:4653-62 PMCID:PMC4985511

[98] Wang R,Li S.MicroRNA-195 suppresses angiogenesis and metastasis of hepatocellular carcinoma by inhibiting the expression of VEGF, VAV2, and CDC42.Hepatology2013;58:642-53

[99] Zeng Y,Hu Z.FGD1 exhibits oncogenic properties in hepatocellular carcinoma through regulating cell morphology, autophagy and mitochondrial function.Biomed Pharmacother2020;125:110029

[100] Martínez-López N,Varela-Rey M.Hepatoma cells from mice deficient in glycine N-methyltransferase have increased RAS signaling and activation of liver kinase B1.Gastroenterology2012;143:787-98.e13 PMCID:PMC3429651

[101] Sun X,Zhang F.Inhibition of NET-1 suppresses proliferation and promotes apoptosis of hepatocellular carcinoma cells by activating the PI3K/AKT signaling pathway.Exp Ther Med2019;17:2334-40 PMCID:PMC6395973

[102] Molee P,Reamtong O.Up-regulation of AKAP13 and MAGT1 on cytoplasmic membrane in progressive hepatocellular carcinoma: a novel target for prognosis.Int J Clin Exp Pathol2015;8:9796-811 PMCID:PMC4637775

[103] Cheng IK,Lai KP.GEF-H1 over-expression in hepatocellular carcinoma promotes cell motility via activation of RhoA signalling.J Pathol2012;228:575-85

[104] Wang B,Qu L,Zhou J.Upregulated TRIO expression correlates with a malignant phenotype in human hepatocellular carcinoma.Tumour Biol2015;36:6901-8

[105] Wong CM,Ching YP,Ng IO.Genetic and epigenetic alterations of DLC-1 gene in hepatocellular carcinoma.Cancer Res2003;63:7646-51

[106] Wong CM,Ching YP.Rho GTPase-activating protein deleted in liver cancer suppresses cell proliferation and invasion in hepatocellular carcinoma.Cancer Res2005;65:8861-8

[107] Ching YP,Chan SF.Deleted in liver cancer (DLC) 2 encodes a RhoGAP protein with growth suppressor function and is underexpressed in hepatocellular carcinoma.J Biol Chem2003;278:10824-30

[108] Xiaorong L,Liyuan Q.Underexpression of deleted in liver cancer 2 (DLC2) is associated with overexpression of RhoA and poor prognosis in hepatocellular carcinoma.BMC Cancer2008;8:205 PMCID:PMC2496915