Mutant GNAQ promotes cell viability and migration of uveal melanoma cells through the activation of Notch signaling

Liu,Honglei; Lei,Chunling; Long,Keqin; Yang,Xinguang; Zhu,Zhaoliang; Zhang,Lihua; Liu,Jun

doi:10.3892/or.2015.3949

Mutant GNAQ promotes cell viability and migration of uveal melanoma cells through the activation of Notch signaling

Authors:
- Honglei Liu
- Chunling Lei
- Keqin Long
- Xinguang Yang
- Zhaoliang Zhu
- Lihua Zhang
- Jun Liu
View Affiliations

Affiliations: Department of Ophthalmology, Xi'an No. 4 Hospital, Xi'an, Shaanxi 710004, P.R. China, Department of Gerontology, Tangdu Hospital, Xi'an, Shaanxi 710038, P.R. China
Published online on: May 5, 2015 https://doi.org/10.3892/or.2015.3949
Pages: 295-301

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

The occurrence of guanine nucleotide binding protein (G protein), q polypeptide (GNAQ) mutations has been found to be high in the majority of uveal melanomas. However, the underlying molecular mechanism of GNAQ mutations in modulating uveal melanoma is poorly understood. The aim of the present study was to investigate the role and underlying mechanism of mutant GNAQ in the regulation of cell viability and migration of uveal melanoma cells. Uveal melanoma cells containing mutant GNAQ were transfected with scrambled or GNAQ small‑interfering RNA. Compared with the control, GNAQ knockdown markedly inhibited cell viability and migration. However, tumor cells without GNAQ mutations exhibited enhanced viability and migration following transfection with HA‑GαqQL. Additionally, GNAQ knockdown significantly downregulated the expression of Jag‑1 (Notch ligand), Notch intracellular domain and Hes‑1 (Notch target gene) in uveal melanoma cells. Conversely, the GNAQ overexpression promoted their expression. Cell viability and migration induced by GNAQ was significantly inhibited following treatment with 5 µmol/l MRK003, a Notch signaling inhibitor. Furthermore, the transfection of human influenza hemagglutinin A epitope (HA)‑GαqQL into tumor cells caused Yes-associated protein (YAP) dephosphorylation and nuclear translocation, which stimulated the expression of Jag‑1 and Hes‑1. Positive correlations were observed between the GNAQ and Jag‑1 mRNA levels and between the GNAQ and Hes‑1 mRNA levels. However, no positive correlation was observed between the GNAQ and YAP mRNA levels. The results suggested that GNAQ mutation induced viability and migration of uveal melanoma cells via Notch signaling activation, which is mediated by YAP dephosphorylation and nuclear translocation.

View Figures

View References

1	Bedikian AY: Metastatic uveal melanoma therapy: Current options. Int Ophthalmol Clin. 46:151–166. 2006. View Article : Google Scholar
2	Biswas J, Krishnakumar S and Shanmugam MP: Uveal melanoma in Asian Indians: A clinicopathological study. Arch Ophthalmol. 120:522–523. 2002.PubMed/NCBI
3	Sakamoto T, Sakamoto M, Yoshikawa H, Hata Y, Ishibashi T, Ohnishi Y and Inomata H: Histologic findings and prognosis of uveal malignant melanoma in Japanese patients. Am J Ophthalmol. 121:276–283. 1996. View Article : Google Scholar : PubMed/NCBI
4	Kuo PK, Puliafito CA, Wang KM, Liu HS and Wu BF: Uveal melanoma in China. Int Ophthalmol Clin. 22:57–71. 1982. View Article : Google Scholar : PubMed/NCBI
5	Singh AD, Turell ME and Topham AK: Uveal melanoma: Trends in incidence, treatment, and survival. Ophthalmology. 118:1881–1885. 2011. View Article : Google Scholar : PubMed/NCBI
6	Collaborative Ocular Melanoma Study Group: Assessment of metastatic disease status at death in 435 patients with large choroidal melanoma in the Collaborative Ocular Melanoma Study (COMS): COMS report no. 15. Arch Ophthalmol. 119:670–676. 2001. View Article : Google Scholar : PubMed/NCBI
7	Gragoudas ES, Egan KM, Seddon JM, Glynn RJ, Walsh SM, Finn SM, Munzenrider JE and Spar MD: Survival of patients with metastases from uveal melanoma. Ophthalmology. 98:383–390. 1991. View Article : Google Scholar : PubMed/NCBI
8	O’Hayre M, Vázquez-Prado J, Kufareva I, Stawiski EW, Handel TM, Seshagiri S and Gutkind JS: The emerging mutational landscape of G proteins and G-protein-coupled receptors in cancer. Nat Rev Cancer. 13:412–424. 2013. View Article : Google Scholar
9	Daniels AB, Lee JE, MacConaill LE, Palescandolo E, Van Hummelen P, Adams SM, DeAngelis MM, Hahn WC, Gragoudas ES, Harbour JW, et al: High throughput mass spectrometry-based mutation profiling of primary uveal melanoma. Invest Ophthalmol Vis Sci. 53:6991–6996. 2012. View Article : Google Scholar : PubMed/NCBI
10	Ambrosini G, Pratilas CA, Qin LX, Tadi M, Surriga O, Carvajal RD and Schwartz GK: Identification of unique MEK-dependent genes in GNAQ mutant uveal melanoma involved in cell growth, tumor cell invasion, and MEK resistance. Clin Cancer Res. 18:3552–3561. 2012. View Article : Google Scholar : PubMed/NCBI
11	Van Raamsdonk CD, Griewank KG, Crosby MB, Garrido MC, Vemula S, Wiesner T, Obenauf AC, Wackernagel W, Green G, Bouvier N, et al: Mutations in GNA11 in uveal melanoma. N Engl J Med. 363:2191–2199. 2010. View Article : Google Scholar : PubMed/NCBI
12	Mumm JS and Kopan R: Notch signaling: From the outside in. Dev Biol. 228:151–165. 2000. View Article : Google Scholar : PubMed/NCBI
13	Brou C, Logeat F, Gupta N, Bessia C, LeBail O, Doedens JR, Cumano A, Roux P, Black RA and Israël A: A novel proteolytic cleavage involved in Notch signaling: The role of the disintegrin-metalloprotease TACE. Mol Cell. 5:207–216. 2000. View Article : Google Scholar : PubMed/NCBI
14	Borggrefe T and Oswald F: The Notch signaling pathway: Transcriptional regulation at Notch target genes. Cell Mol Life Sci. 66:1631–1646. 2009. View Article : Google Scholar : PubMed/NCBI
15	Jarriault S, Le Bail O, Hirsinger E, Pourquié O, Logeat F, Strong CF, Brou C, Seidah NG and Isral A: Delta-1 activation of notch-1 signaling results in HES-1 transactivation. Mol Cell Biol. 18:7423–7431. 1998.PubMed/NCBI
16	Liu ZJ, Xiao M, Balint K, Smalley KS, Brafford P, Qiu R, Pinnix CC, Li X and Herlyn M: Notch1 signaling promotes primary melanoma progression by activating mitogen-activated protein kinase/phosphatidylinositol 3-kinase-Akt pathways and up-regulating N-cadherin expression. Cancer Res. 66:4182–4190. 2006. View Article : Google Scholar : PubMed/NCBI
17	Pinnix CC, Lee JT, Liu ZJ, McDaid R, Balint K, Beverly LJ, Brafford PA, Xiao M, Himes B, Zabierowski SE, et al: Active Notch1 confers a transformed phenotype to primary human melanocytes. Cancer Res. 69:5312–5320. 2009. View Article : Google Scholar : PubMed/NCBI
18	Asnaghi L, Ebrahimi KB, Schreck KC, Bar EE, Coonfield ML, Bell WR, Handa J, Merbs SL, Harbour JW and Eberhart CG: Notch signaling promotes growth and invasion in uveal melanoma. Clin Cancer Res. 18:654–665. 2012. View Article : Google Scholar : PubMed/NCBI
19	Babchia N, Calipel A, Mouriaux F, Faussat AM and Mascarelli F: The PI3K/Akt and mTOR/P70S6K signaling pathways in human uveal melanoma cells: Interaction with B-Raf/ERK. Invest Ophthalmol Vis Sci. 51:421–429. 2010. View Article : Google Scholar
20	Mende U, Kagen A, Cohen A, Aramburu J, Schoen FJ and Neer EJ: Transient cardiac expression of constitutively active Gα_q leads to hypertrophy and dilated cardiomyopathy by calci-neurin-dependent and independent pathways. Proc Natl Acad Sci USA. 95:13893–13898. 1998. View Article : Google Scholar
21	Marinissen MJ, Servitja JM, Offermanns S, Simon MI and Gutkind JS: Thrombin protease-activated receptor-1 signals through G_q- and G₁₃-initiated MAPK cascades regulating c-Jun expression to induce cell transformation. J Biol Chem. 278:46814–46825. 2003. View Article : Google Scholar : PubMed/NCBI
22	Yu FX, Luo J, Mo JS, Liu G, Kim YC, Meng Z, Zhao L, Peyman G, Ouyang H, Jiang W, et al: Mutant Gq/11 promote uveal melanoma tumorigenesis by activating YAP. Cancer Cell. 25:822–830. 2014. View Article : Google Scholar : PubMed/NCBI
23	Cheng P, Kumar V, Liu H, Youn JI, Fishman M, Sherman S and Gabrilovich D: Effects of notch signaling on regulation of myeloid cell differentiation in cancer. Cancer Res. 74:141–152. 2014. View Article : Google Scholar
24	Neradugomma NK, Subramaniam D, Tawfik OW, Goffin V, Kumar TR, Jensen RA and Anant S: Prolactin signaling enhances colon cancer stemness by modulating Notch signaling in a Jak2-STAT3/ERK manner. Carcinogenesis. 35:795–806. 2014. View Article : Google Scholar :
25	Xie M, He CS, Wei SH and Zhang L: Notch-1 contributes to epidermal growth factor receptor tyrosine kinase inhibitor acquired resistance in non-small cell lung cancer in vitro and in vivo. Eur J Cancer. 49:3559–3572. 2013. View Article : Google Scholar : PubMed/NCBI
26	Lamba S, Felicioni L, Buttitta F, Bleeker FE, Malatesta S, Corbo V, Scarpa A, Rodolfo M, Knowles M, Frattini M, et al: Mutational profile of GNAQ^Q209 in human tumors. PLoS One. 4:e68332009. View Article : Google Scholar
27	Tschaharganeh DF, Chen X, Latzko P, Malz M, Gaida MM, Felix K, Ladu S, Singer S, Pinna F, Gretz N, et al: Yes-associated protein up-regulates Jagged-1 and activates the Notch pathway in human hepatocellular carcinoma. Gastroenterology. 144:1530–1542.e12. 2013. View Article : Google Scholar : PubMed/NCBI
28	Kan Z, Jaiswal BS, Stinson J, Janakiraman V, Bhatt D, Stern HM, Yue P, Haverty PM, Bourgon R, Zheng J, et al: Diverse somatic mutation patterns and pathway alterations in human cancers. Nature. 466:869–873. 2010. View Article : Google Scholar : PubMed/NCBI
29	Prickett TD, Wei X, Cardenas-Navia I, Teer JK, Lin JC, Walia V, Gartner J, Jiang J, Cherukuri PF, Molinolo A, et al: Exon capture analysis of G protein-coupled receptors identifies activating mutations in GRM3 in melanoma. Nat Genet. 43:1119–1126. 2011. View Article : Google Scholar : PubMed/NCBI
30	Küsters-Vandevelde HV, Klaasen A, Küsters B, Groenen PJ, van Engen-van Grunsven IA, van Dijk MR, Reifenberger G, Wesseling P and Blokx WA: Activating mutations of the GNAQ gene: A frequent event in primary melanocytic neoplasms of the central nervous system. Acta Neuropathol. 119:317–323. 2010. View Article : Google Scholar :
31	Van Raamsdonk CD, Bezrookove V, Green G, Bauer J, Gaugler L, O’Brien JM, Simpson EM, Barsh GS and Bastian BC: Frequent somatic mutations of GNAQ in uveal melanoma and blue naevi. Nature. 457:599–602. 2009. View Article : Google Scholar :
32	Vallar L, Spada A and Giannattasio G: Altered G_s and adenylate cyclase activity in human GH-secreting pituitary adenomas. Nature. 330:566–568. 1987. View Article : Google Scholar : PubMed/NCBI
33	Lyons J, Landis CA, Harsh G, Vallar L, Grünewald K, Feichtinger H, Duh QY, Clark OH, Kawasaki E, Bourne HR, et al: Two G protein oncogenes in human endocrine tumors. Science. 249:655–659. 1990. View Article : Google Scholar : PubMed/NCBI
34	Van Raamsdonk CD, Fitch KR, Fuchs H, de Angelis MH and Barsh GS: Effects of G-protein mutations on skin color. Nat Genet. 36:961–968. 2004. View Article : Google Scholar : PubMed/NCBI
35	Markby DW, Onrust R and Bourne HR: Separate GTP binding and GTPase activating domains of a G alpha subunit. Science. 262:1895–1901. 1993. View Article : Google Scholar : PubMed/NCBI
36	McCubrey JA, Steelman LS, Chappell WH, Abrams SL, Wong EW, Chang F, Lehmann B, Terrian DM, Milella M, Tafuri A, et al: Roles of the Raf/MEK/ERK pathway in cell growth, malignant transformation and drug resistance. Biochim Biophys Acta. 1773:1263–1284. 2007. View Article : Google Scholar
37	Hoek K, Rimm DL, Williams KR, Zhao H, Ariyan S, Lin A, Kluger HM, Berger AJ, Cheng E, Trombetta ES, et al: Expression profiling reveals novel pathways in the transformation of melanocytes to melanomas. Cancer Res. 64:5270–5282. 2004. View Article : Google Scholar : PubMed/NCBI
38	Qin JZ, Stennett L, Bacon P, Bodner B, Hendrix MJ, Seftor RE, Seftor EA, Margaryan NV, Pollock PM, Curtis A, et al: p53-independent NOXA induction overcomes apoptotic resistance of malignant melanomas. Mol Cancer Ther. 3:895–902. 2004.PubMed/NCBI
39	Asnaghi L, Handa JT, Merbs SL, Harbour JW and Eberhart CG: A role for Jag2 in promoting uveal melanoma dissemination and growth. Invest Ophthalmol Vis Sci. 54:295–306. 2013. View Article : Google Scholar
40	Huang X, Wang L, Zhang H, Wang H, Zhao X, Qian G, Hu J, Ge S and Fan X: Therapeutic efficacy by targeting correction of Notch1-induced aberrants in uveal tumors. PLoS One. 7:e443012012. View Article : Google Scholar : PubMed/NCBI
41	Zhou D, Zhang Y, Wu H, Barry E, Yin Y, Lawrence E, Dawson D, Willis JE, Markowitz SD, Camargo FD, et al: Mst1 and Mst2 protein kinases restrain intestinal stem cell proliferation and colonic tumorigenesis by inhibition of Yes-associated protein (Yap) overabundance. Proc Natl Acad Sci USA. 108:E1312–E1320. 2011. View Article : Google Scholar : PubMed/NCBI
42	Feng X, Degese MS, Iglesias-Bartolome R, Vaque JP, Molinolo AA, Rodrigues M, Zaidi MR, Ksander BR, Merlino G, Sodhi A, et al: Hippo-independent activation of YAP by the GNAQ uveal melanoma oncogene through a trio-regulated rho GTPase signaling circuitry. Cancer Cell. 25:831–845. 2014. View Article : Google Scholar : PubMed/NCBI