Abstract
Ovarian cancer is the most lethal gynecological malignancy and molecular mechanisms of its progression and metastasis are not completely understood. Some members of GAB (GRB2-associated binding) protein family have been reported to be involved in tumor cell proliferation and metastasis in various cancer types. In the present study, we analyzed the expression of GAB proteins (GAB1, GAB2 and GAB3) in ovarian cancer compared to normal ovarian tissue, in terms of tumor stage, tumor grade and histological type. Differential expression analyses performed in R programming environment using multiple transcriptome datasets (n = 1449) showed that GAB1 expression is decreased in ovarian cancer independently of tumor stage, grade and histotype. Unlike GAB1, expression of GAB2 and GAB3 are increased from early stage to late stage and from low grade to high grade in epithelial ovarian cancer. GAB2 and GAB3 also showed histotype-dependent expression. GAB3 was computed as a top gene whose expression most significantly changed between tumor cells from primary tumor, metastases and ascites. High expression of GAB2 and GAB3 was shown to be associated with shorter progression-free survival in ovarian cancer. This study shows that GAB2 and GAB3 can be important regulators of tumor progression and metastasis in ovarian cancer.
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Transcriptomics datasets used in the present study are available from Gene Expression Omnibus (GEO) and TCGA-OV (The Cancer Genome Atlas – Ovarian Cancer), and their accession IDs were given in the text. Most expression data can be accessed in R programming environment using curatedOvarianData Bioconductor data package. Protein datasets used in the study and information on how to access them were given in Materials and Methods section.
References
Allaire, JJ, Xie, Y, McPherson, J, Luraschi, J, Ushey, K, Atkins, A, Wickham, H, Cheng, J, Chang, W and Iannone, R (2020). Rmarkdown: dynamic documents for R. R package version 2.2. URL https://rmarkdown.rstudio.com
Auguie, B (2017). gridExtra: miscellaneous functions for “grid” graphics. R package version 2.3. https://CRAN.R-project.org/package=gridExtra
Barretina J, Caponigro G, Stransky N, Venkatesan K, Margolin AA, Kim S, Wilson CJ, Lehár J, Kryukov GV, Sonkin D, Reddy A, Liu M, Murray L, Berger MF, Monahan JE, Morais P, Meltzer J, Korejwa A, Jané-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, Jagtap K, Jones MD, Wang L, Hatton C, Palescandolo E, Gupta S, Mahan S, Sougnez C, Onofrio RC, Liefeld T, MacConaill L, Winckler W, Reich M, Li N, Mesirov JP, Gabriel SB, Getz G, Ardlie K, Chan V, Myer VE, Weber BL, Porter J, Warmuth M, Finan P, Harris JL, Meyerson M, Golub TR, Morrissey MP, Sellers WR, Schlegel R, Garraway LA (2012) The Cancer cell line encyclopedia enables predictive modelling of anticancer drug sensitivity. Nature 483(7391):603–607. https://doi.org/10.1038/nature11003
Bentires-Alj M, Gil SG, Chan R, Wang ZC, Wang Y, Imanaka N, Harris LN, Richardson A, Neel BG, Gu H (2006) A role for the scaffolding adapter GAB2 in breast cancer. Nat Med 12(1):114–121. https://doi.org/10.1038/nm1341
Berkel C, Cacan E (2020) In silico analysis of DYNLL1 expression in ovarian cancer chemoresistance. Cell Biol Int, Advance online publication 44:1598–1605. https://doi.org/10.1002/cbin.11352
Bonome T, Levine DA, Shih J, Randonovich M, Pise-Masison CA, Bogomolniy F, Ozbun L, Brady J, Barrett JC, Boyd J, Birrer MJ (2008) A gene signature predicting for survival in suboptimally debulked patients with ovarian cancer. Cancer Res 68(13):5478–5486. https://doi.org/10.1158/0008-5472.CAN-07-6595
Brown LA, Kalloger SE, Miller MA, Shih I, McKinney SE, Santos JL, Swenerton K, Spellman PT, Gray J, Gilks CB, Huntsman DG (2008) Amplification of 11q13 in ovarian carcinoma. Genes Chromosom Cancer 47(6):481–489. https://doi.org/10.1002/gcc.20549
Cacan E (2016) Histone Deacetylase-1-mediated suppression of FAS in Chemoresistant ovarian Cancer cells. Anticancer Res 36(6):2819–2826
Cacan E, Ali MW, Boyd NH, Hooks SB, Greer SF (2014) Inhibition of HDAC1 and DNMT1 modulate RGS10 expression and decrease ovarian cancer chemoresistance. PLoS One 9(1):e87455. https://doi.org/10.1371/journal.pone.0087455
Cancer Genome Atlas Research Network (2011) Integrated genomic analyses of ovarian carcinoma. Nature 474(7353):609–615. https://doi.org/10.1038/nature10166
Chen D, Cao L, Wang X (2019) MPZL1 promotes tumor cell proliferation and migration via activation of Src kinase in ovarian cancer. Oncol Rep 42(2):679–687. https://doi.org/10.3892/or.2019.7199
Davis S, Meltzer PS (2007) GEOquery: a bridge between the gene expression omnibus (GEO) and BioConductor. Bioinformatics (Oxford, England) 23(14):1846–1847. https://doi.org/10.1093/bioinformatics/btm254
Duckworth C, Zhang L, Carroll SL, Ethier SP, Cheung HW (2016) Overexpression of GAB2 in ovarian cancer cells promotes tumor growth and angiogenesis by upregulating chemokine expression. Oncogene 35(31):4036–4047. https://doi.org/10.1038/onc.2015.472
Dunn GP, Cheung HW, Agarwalla PK, Thomas S, Zektser Y, Karst AM, Boehm JS, Weir BA, Berlin AM, Zou L, Getz G, Liu JF, Hirsch M, Vazquez F, Root DE, Beroukhim R, Drapkin R, Hahn WC (2014) In vivo multiplexed interrogation of amplified genes identifies GAB2 as an ovarian cancer oncogene. Proc Natl Acad Sci U S A 111(3):1102–1107. https://doi.org/10.1073/pnas.1311909111
Eckert MA, Coscia F, Chryplewicz A, Chang JW, Hernandez KM, Pan S, Tienda SM, Nahotko DA, Li G, Blaženović I, Lastra RR, Curtis M, Yamada SD, Perets R, McGregor SM, Andrade J, Fiehn O, Moellering RE, Mann M, Lengyel E (2019) Proteomics reveals NNMT as a master metabolic regulator of cancer-associated fibroblasts. Nature 569(7758):723–728. https://doi.org/10.1038/s41586-019-1173-8
Fan G, Zhang S, Gao Y, Greer PA, Tonks NK (2016) HGF-independent regulation of MET and GAB1 by nonreceptor tyrosine kinase FER potentiates metastasis in ovarian cancer. Genes Dev 30(13):1542–1557. https://doi.org/10.1101/gad.284166.116
Fang Z, Li T, Chen W, Wu D, Qin Y, Liu M, Wu G, He L, Li H, Gu H (2019) Gab2 promotes cancer stem cell like properties and metastatic growth of ovarian cancer via downregulation of miR-200c. Exp Cell Res 382(1):111462. https://doi.org/10.1016/j.yexcr.2019.06.007
Ferlay J, Soerjomataram I, Dikshit R, Eser S, Mathers C, Rebelo M, Parkin DM, Forman D, Bray F (2015) Cancer incidence and mortality worldwide: sources, methods and major patterns in GLOBOCAN 2012. Int J Cancer 136(5):E359–E386. https://doi.org/10.1002/ijc.29210
Ganzfried BF, Riester M, Haibe-Kains B, Risch T, Tyekucheva S, Jazic I, Wang XV, Ahmadifar M, Birrer MJ, Parmigiani G, Huttenhower C, Waldron L (2013) curatedOvarianData: clinically annotated data for the ovarian cancer transcriptome. Database : the journal of biological databases and curation, 2013, bat013. https://doi.org/10.1093/database/bat013
Gao Q, Yang Z, Xu S, Li X, Yang X, Jin P, Liu Y, Zhou X, Zhang T, Gong C, Wei X, Liu D, Sun C, Chen G, Hu J, Meng L, Zhou J, Sawada K, Fruscio R, Grunt TW, Wischhusen J, Vargas-Hernández VM, Pothuri B, Coleman RL (2019) Heterotypic CAF-tumor spheroids promote early peritoneal metastatis of ovarian cancer. J Exp Med 216(3):688–703. https://doi.org/10.1084/jem.20180765
Grither WR & Longmore GD (2018) Inhibition of tumor-microenvironment interaction and tumor invasion by small-molecule allosteric inhibitor of DDR2 extracellular domain. PNAS 115(33), E7786–E7794. https://doi.org/10.1073/pnas.1805020115
Gyorffy B, Lánczky A, Szállási Z (2012) Implementing an online tool for genome-wide validation of survival-associated biomarkers in ovarian-cancer using microarray data from 1287 patients. Endocr Relat Cancer 19(2):197–208. https://doi.org/10.1530/ERC-11-0329
Hendrix ND, Wu R, Kuick R, Schwartz DR, Fearon ER, Cho KR (2006) Fibroblast growth factor 9 has oncogenic activity and is a downstream target of Wnt signaling in ovarian endometrioid adenocarcinomas. Cancer Res 66(3):1354–1362. https://doi.org/10.1158/0008-5472.CAN-05-3694
Hoeben A, Martin D, Clement PM, Cools J, Gutkind JS (2013) Role of GRB2-associated binder 1 in epidermal growth factor receptor-induced signaling in head and neck squamous cell carcinoma. Int J Cancer 132(5):1042–1050. https://doi.org/10.1002/ijc.27763
Horst B, Gruvberger-Saal SK, Hopkins BD, Bordone L, Yang Y, Chernoff KA, Uzoma I, Schwipper V, Liebau J, Nowak NJ, Brunner G, Owens D, Rimm DL, Parsons R, Celebi JT (2009) Gab2-mediated signaling promotes melanoma metastasis. Am J Pathol 174(4):1524–1533. https://doi.org/10.2353/ajpath.2009.080543
Hu Z, Li J, Gao Q, Wei S, Yang B (2017) SHP2 overexpression enhances the invasion and metastasis of ovarian cancer in vitro and in vivo. OncoTargets and therapy 10:3881–3891. https://doi.org/10.2147/OTT.S138833
Huang D, Sherman BT, Lempicki RA (2009a) Bioinformatics enrichment tools: paths toward the comprehensive functional analysis of large gene lists. Nucleic Acids Res 37(1):1–13. https://doi.org/10.1093/nar/gkn923
Huang D, Sherman BT, Lempicki RA (2009b) Systematic and integrative analysis of large gene lists using DAVID bioinformatics resources. Nat Protoc 4(1):44–57. https://doi.org/10.1038/nprot.2008.211
Huang Y, Hu Y, Jin Z, Shen Z (2018) LncRNA snaR upregulates GRB2-associated binding protein 2 and promotes proliferation of ovarian carcinoma cells. Biochem Biophys Res Commun 503(3):2028–2032. https://doi.org/10.1016/j.bbrc.2018.07.152
Huber W, Carey VJ, Gentleman R, Anders S, Carlson M, Carvalho BS, Bravo HC, Davis S, Gatto L, Girke T, Gottardo R, Hahne F, Hansen KD, Irizarry RA, Lawrence M, Love MI, MacDonald J, Obenchain V, Oleś AK, Pagès H, Reyes A, Shannon P, Smyth GK, Tenenbaum D, Waldron L, Morgan M (2015) Orchestrating high-throughput genomic analysis with Bioconductor. Nat Methods 12(2):115–121. https://doi.org/10.1038/nmeth.3252
Jia P, Li F, Gu W, Zhang W, Cai Y (2017) Gab3 overexpression in human glioma mediates Akt activation and tumor cell proliferation. PLoS One 12(3):e0173473. https://doi.org/10.1371/journal.pone.0173473
Kanlikilicer P, Ozpolat B, Aslan B, Bayraktar R, Gurbuz N, Rodriguez-Aguayo C, Bayraktar E, Denizli M, Gonzalez-Villasana V, Ivan C, Lokesh G, Amero P, Catuogno S, Haemmerle M, Wu SY, Mitra R, Gorenstein DG, Volk DE, de Franciscis V, Sood AK et al (2017) Therapeutic targeting of AXL receptor tyrosine kinase inhibits tumor growth and Intraperitoneal metastasis in ovarian Cancer models. Molecular therapy Nucleic acids 9:251–262. https://doi.org/10.1016/j.omtn.2017.06.023
Karlan BY, Dering J, Walsh C, Orsulic S, Lester J, Anderson LA, Ginther CL, Fejzo M, Slamon D (2014) POSTN/TGFBI-associated stromal signature predicts poor prognosis in serous epithelial ovarian cancer. Gynecol Oncol 132(2):334–342. https://doi.org/10.1016/j.ygyno.2013.12.021
Kassambara A (2020) ggpubr: 'ggplot2' Based Publication Ready Plots. R package version 0.4.0. https://CRAN.R-project.org/package=ggpubr
Lara OD, Bayraktar E, Amero P, Ma S, Ivan C, Hu W, Wang Y, Mangala LS, Dutta P, Bhattacharya P, Ashizawa AT, Lopez-Berestein G, Rodriguez-Aguayo C, Sood AK (2020) Therapeutic efficacy of liposomal Grb2 antisense oligodeoxynucleotide (L-Grb2) in preclinical models of ovarian and uterine cancer. Oncotarget 11(29):2819–2833. https://doi.org/10.18632/oncotarget.27667
Li J, Zhao W, Akbani R, Liu W, Ju Z, Ling S, Vellano CP, Roebuck P, Yu Q, Eterovic AK, Byers LA, Davies MA, Deng W, Gopal YN, Chen G, von Euw EM, Slamon D, Conklin D, Heymach JV, Gazdar AF et al (2017) Characterization of human Cancer cell lines by reverse-phase protein arrays. Cancer Cell 31(2):225–239. https://doi.org/10.1016/j.ccell.2017.01.005
Mok SC, Bonome T, Vathipadiekal V, Bell A, Johnson ME, Wong KK, Park DC, Hao K, Yip DK, Donninger H, Ozbun L, Samimi G, Brady J, Randonovich M, Pise-Masison CA, Barrett JC, Wong WH, Welch WR, Berkowitz RS, Birrer MJ (2009) A gene signature predictive for outcome in advanced ovarian cancer identifies a survival factor: microfibril-associated glycoprotein 2. Cancer Cell 16(6):521–532. https://doi.org/10.1016/j.ccr.2009.10.018
Ooms, J (2020a). Magick: advanced graphics and image-processing in R. R package version 2.3. https://CRAN.R-project.org/package=magick
Ooms, J (2020b). Pdftools: text extraction, rendering and converting of PDF documents. R package version 2.3.1. https://CRAN.R-project.org/package=pdftools
Oughtred R, Stark C, Breitkreutz BJ, Rust J, Boucher L, Chang C, Kolas N, O'Donnell L, Leung G, McAdam R, Zhang F, Dolma S, Willems A, Coulombe-Huntington J, Chatr-Aryamontri A, Dolinski K, Tyers M (2019) The BioGRID interaction database: 2019 update. Nucleic Acids Res 47(D1):D529–D541. https://doi.org/10.1093/nar/gky1079
R Core Team (2020). R: A language and environment for statistical computing. R Foundation for statistical computing, Vienna, Austria. URL https://www.R-project.org/
Ritchie, ME, Phipson, B, Wu, D, Hu, Y, Law, CW, Shi, W and Smyth, GK (2015). Limma powers differential expression analyses for RNA-sequencing and microarray studies. Nucleic acids research, 43(7), e47. https://doi.org/10.1093/nar/gkv00730.Kassambara, A. (2020). Ggpubr: ‘ggplot2’ based publication ready plots. R package version 0.3.0. https://CRAN.R-project.org/package=ggpubr
Seiden-Long I, Navab R, Shih W, Li M, Chow J, Zhu CQ, Radulovich N, Saucier C, Tsao MS (2008) Gab1 but not Grb2 mediates tumor progression in met overexpressing colorectal cancer cells. Carcinogenesis 29(3):647–655. https://doi.org/10.1093/carcin/bgn009
Tothill RW, Tinker AV, George J, Brown R, Fox SB, Lade S, Johnson DS, Trivett MK, Etemadmoghadam D, Locandro B, Traficante N, Fereday S, Hung JA, Chiew YE, Haviv I, Australian Ovarian Cancer Study Group, Gertig D, DeFazio A, Bowtell DD (2008) Novel molecular subtypes of serous and endometrioid ovarian cancer linked to clinical outcome. Clinical cancer research : an official journal of the American Association for Cancer Research 14(16):5198–5208. https://doi.org/10.1158/1078-0432.CCR-08-0196
Uhlén, M, Fagerberg, L, Hallström, BM, Lindskog, C, Oksvold, P, Mardinoglu, A, Sivertsson, Å, Kampf, C, Sjöstedt, E, Asplund, A, Olsson, I, Edlund, K, Lundberg, E, Navani, S, Szigyarto, CA, Odeberg, J, Djureinovic, D, Takanen, JO, Hober, S, Alm, T, … Pontén, F (2015). Proteomics. Tissue-based map of the human proteome. Science (New York, N.Y.), 347(6220), 1260419. https://doi.org/10.1126/science.1260419
Uhlén M, Zhang C, Lee S, Sjöstedt E, Fagerberg L, Bidkhori G, Benfeitas R, Arif M, Liu Z, Edfors F, Sanli K, von Feilitzen K, Oksvold P, Lundberg E, Hober S, Nilsson P, Mattsson J, Schwenk JM, Brunnström H, Glimelius B et al (2017) A pathology atlas of the human cancer transcriptome. Science (New York, N.Y.) 357(6352):eaan2507. https://doi.org/10.1126/science.aan2507
Wang Y, Sheng Q, Spillman MA, Behbakht K, Gu H (2012) Gab2 regulates the migratory behaviors and E-cadherin expression via activation of the PI3K pathway in ovarian cancer cells. Oncogene 31(20):2512–2520. https://doi.org/10.1038/onc.2011.435
Wang X, Peng J, Yang Z, Zhou PJ, An N, Wei L, Zhu HH, Lu J, Fang YX, Gao WQ (2019) Elevated expression of Gab1 promotes breast cancer metastasis by dissociating the PAR complex. Journal of experimental & clinical cancer research : CR 38(1):27. https://doi.org/10.1186/s13046-019-1025-2
Wickham, H and Bryan, J. (2019). Readxl: read excel files. R package version 1.3.1. https://CRAN.R-project.org/package=readxl
Wickham H, Averick M, Bryan J, Chang W, D’Agostino McGowan L, François R, Grolemund G, Hayes A, Henry L, Hester J, Kuhn M, Lin Pedersen T, Miller E, Bache SM, Müller K, Ooms J, Robinson D, Seidel DP, Spinu V, Takahashi K, Vaughan D, Wilke C, Woo K, Yutani H (2019) Welcome to the tidyverse. Journal of open source software 4(43):1686. https://doi.org/10.21105/joss.01686
Wöhrle FU, Daly RJ, Brummer T (2009) Function, regulation and pathological roles of the gab/DOS docking proteins. Cell communication and signaling : CCS 7:22. https://doi.org/10.1186/1478-811X-7-22
Wu AY, Gu LY, Cang W, Cheng MX, Wang WJ, Di W, Huang L, Qiu LH (2019) Fn14 overcomes cisplatin resistance of high-grade serous ovarian cancer by promoting Mdm2-mediated p53-R248Q ubiquitination and degradation. Journal of experimental & clinical cancer research : CR 38(1):176. https://doi.org/10.1186/s13046-019-1171-6
Xiang S, Wang N, Hui P, Ma J (2017) Gab3 is required for human colorectal cancer cell proliferation. Biochem Biophys Res Commun 484(4):719–725. https://doi.org/10.1016/j.bbrc.2017.01.095
Xie, Y (2020). Knitr: A general-purpose package for dynamic report generation in R. R package version 1.28
Yang L, Zhang X, Ma Y, Zhao X, Li B, Wang H (2017) Ascites promotes cell migration through the repression of miR-125b in ovarian cancer. Oncotarget 8(31):51008–51015. https://doi.org/10.18632/oncotarget.16846
Yoshihara K, Tajima A, Komata D, Yamamoto T, Kodama S, Fujiwara H, Suzuki M, Onishi Y, Hatae M, Sueyoshi K, Fujiwara H, Kudo Y, Inoue I, Tanaka K (2009) Gene expression profiling of advanced-stage serous ovarian cancers distinguishes novel subclasses and implicates ZEB2 in tumor progression and prognosis. Cancer Sci 100(8):1421–1428. https://doi.org/10.1111/j.1349-7006.2009.01204.x
Zhao G, Chen J, Deng Y, Gao F, Zhu J, Feng Z, Lv X, Zhao Z (2011) Identification of NDRG1-regulated genes associated with invasive potential in cervical and ovarian cancer cells. Biochem Biophys Res Commun 408(1):154–159. https://doi.org/10.1016/j.bbrc.2011.03.140
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Caglar Berkel is funded by TUBITAK (The Scientific and Technological Research Council of Turkey) 2211-E graduate program.
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Berkel, C., Cacan, E. GAB2 and GAB3 are expressed in a tumor stage-, grade- and histotype-dependent manner and are associated with shorter progression-free survival in ovarian cancer. J. Cell Commun. Signal. 15, 57–70 (2021). https://doi.org/10.1007/s12079-020-00582-3
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DOI: https://doi.org/10.1007/s12079-020-00582-3