The role of p19 and p21 H-Ras proteins and mutants in miRNA expression in cancer and a Costello syndrome cell model

BMC Medical Genetics

Montse Bach-Elias²  Carlo M. Croce¹  Cristian Taccioli¹  Stefano Volinia¹  Chang-Gong Liu¹  George A. Calin³  Maria Camats²  Roseli García-Cruz² 

[1] Ohio State University, Department of Molecular Immunology, Virology and Molecular Genetics, Columbus 43210, Ohio, USA;Instituto de Investigaciones Biomédicas de Barcelona- CSIC, C/ Egipcíacas15, Barcelona, 08001, Spain;Present address: Departments of Experimental Therapeutics & Cancer Genetics, University of Texas, MD Anderson Cancer Center, Houston 77030, TX, USA

关键词: H-ras mutants;    Costello syndrome;    miRNAs;    p21;    p19;    H-ras;    IDX;    Alternative splicing;   

Others  :  1220334
DOI  :  10.1186/s12881-015-0184-z

 received in 2015-01-21, accepted in 2015-06-01,  发布年份 2015

【 摘 要 】

Background

P19 H-Ras, a second product derived from the H-Ras gene by alternative splicing, induces a G1/S phase delay, thereby maintaining cells in a reversible quiescence state. When P21 H-Ras is mutated in tumour cells, the alternative protein P19 H-Ras is also mutated. The H-Ras mutation Q61L is frequently detected in different tumours, which acts as constitutive activator of Ras functions and is considered to be a strong activating mutant. Additionally, a rare congenital disorder named Costello Syndrome, is described as a H-Ras disorder in children, mainly due to mutation G12S in p19 and p21 H-Ras proteins, which is present in 90 % of the Costello Syndrome patients. Our aim is to better understand the role of p19 and p21 H-Ras proteins in the cancer and Costello Syndrome development, concerning the miRNAs expression.

Methods

Total miRNAs expression regulated by H-Ras proteins were first analyzed in human miRNA microarrays assays. Previously selected miRNAs, were further analyzed in developed cell lines containing H-Ras protein mutants, that included the G12S Costello Syndrome mutant, with PCR Real-Time Taq Man miRNA Assays primers.

Results

This study describes how p19 affects the RNA world and shows that: i) miR-342, miR-206, miR-330, miR-138 and miR-99b are upregulated by p19 but not by p19W164A mutant; ii) anti-miR-206 can restore the G2 phase in the presence of p19; iii) p19 and p21Q61L regulate their own alternative splicing; iv) miR-206 and miR-138 are differentially regulated by p19 and p21 H-Ras and v) P19G12S Costello mutants show a clear upregulation of miR-374, miR-126, miR-342, miR-330, miR-335 and let-7.

Conclusions

These results allow us to conclude that the H-Ras G12S mutation plays an important role in miRNA expression and open up a new line of study to understand the consequences of this mutation on Costello syndrome. Furthermore, they suggest that oncogenes may have a sufficiently important impact on miRNA expression to promote the development of numerous cancers.

【 授权许可】

   
2015 García-Cruz et al.

【 图 表 】

Fig. 1.

Fig. 2.

Fig. 3.

Fig. 4.

Fig. 5.

【 参考文献 】

• [1]Lowy DR, Willumsen BM. Function and regulation of ras. Annu Rev Biochem. 1993; 62(3):851-91.
• [2]Wennerberg K, Rossman KL, Der CJ. The Ras superfamily at a glance. J Cell Sci. 2005; 118(Pt 5):843-6.
• [3]Takashima A, Faller DV. Targeting the RAS oncogene. Expert Opin Ther Targets. 2013; 17(5):507-31.
• [4]HernandezAlcoceba R, del Peso L, Lacal JC. The Ras family of GTPases in cancer cell invasion. Cell Mol Life Sci. 2000; 57(1):65-76.
• [5]Cohen JB, Broz SD, Levinson AD. Expression of the H-ras proto-oncogene is controlled by alternative splicing. Cell. 1989; 58(3):461-72.
• [6]Guil S, de La Iglesia N, Fernandez-Larrea J, Cifuentes D, Ferrer JC, Guinovart JJ et al.. Alternative splicing of the human proto-oncogene c-H-ras renders a new Ras family protein that trafficks to cytoplasm and nucleus. Cancer Res. 2003; 63(17):5178-87.
• [7]Camats M, Kokolo M, Heesom KJ, Ladomery M, Bach-Elias M. P19 H-ras induces G1/S phase delay maintaining cells in a reversible quiescence state. PLoS One. 2009; 4(12):e8513.
• [8]Kim JW, Kim WH, Jeong MH, Jang SM, Song KH, Park SI et al.. p19(ras) amplifies p73beta-induced apoptosis through mitochondrial pathway. Biochem Biophys Res Commun. 2008; 373(1):146-50.
• [9]Jang SM, Kim JW, Kim CH, Kim D, Rhee S, Choi KH. p19(ras) Represses proliferation of non-small cell lung cancer possibly through interaction with Neuron-Specific Enolase (NSE). Cancer Lett. 2010; 289(1):91-8.
• [10]Cohen JB, Levinson AD. A point mutation in the last intron responsible for increased expression and transforming activity of the c-Ha-ras oncogene. Nature. 1988; 334(6178):119-24.
• [11]Muraoka S, Shima F, Araki M, Inoue T, Yoshimoto A, Ijiri Y et al.. Crystal structures of the state 1 conformations of the GTP-bound H-Ras protein and its oncogenic G12V and Q61L mutants. FEBS Lett. 2012; 586(12):1715-8.
• [12]Yang D, Wang MT, Tang Y, Chen Y, Jiang H, Jones TT et al.. Impairment of mitochondrial respiration in mouse fibroblasts by oncogenic H-RAS(Q61L). Cancer Biol Ther. 2010; 9(2):122-33.
• [13]Lin AE, Rauen KA, Gripp KW, Carey JC. Clarification of previously reported Costello syndrome patients. Am J Med Genet A. 2008; 146(7):940-3.
• [14]Gripp KW, Lin AE, Stabley DL, Nicholson L, Scott CI, Doyle D et al.. HRAS mutation analysis in Costello syndrome: genotype and phenotype correlation. Am J Med Genet A. 2006; 140(1):1-7.
• [15]Gripp KW, Stabley DL, Nicholson L, Hoffman JD, Sol-Church K. Somatic mosaicism for an HRAS mutation causes Costello syndrome. Am J Med Genet A. 2006; 140(20):2163-9.
• [16]Sol-Church K, Stabley DL, Nicholson L, Gonzalez IL, Gripp KW. Paternal bias in parental origin of HRAS mutations in Costello syndrome. Hum Mutat. 2006; 27(8):736-41.
• [17]Niemeyer CM. RAS diseases in children. Haematologica. 2014; 99(11):1653-62.
• [18]Morice-Picard F, Ezzedine K, Delrue MA, Arveiler B, Fergelot P, Taieb A et al.. Cutaneous manifestations in Costello and cardiofaciocutaneous syndrome: report of 18 cases and literature review. Pediatr Dermatol. 2013; 30(6):665-73.
• [19]Felekkis K, Touvana E, Stefanou C, Deltas C. microRNAs: a newly described class of encoded molecules that play a role in health and disease. Hippokratia. 2010; 14(4):236-40.
• [20]Castellano E, De Las RJ, Guerrero C, Santos E. Transcriptional networks of knockout cell lines identify functional specificities of H-Ras and N-Ras: significant involvement of N-Ras in biotic and defense responses. Oncogene. 2007; 26(6):917-33.
• [21]Liu CG, Calin GA, Meloon B, Gamliel N, Sevignani C, Ferracin M et al.. An oligonucleotide microchip for genome-wide microRNA profiling in human and mouse tissues. Proc Natl Acad Sci U S A. 2004; 101(26):9740-4.
• [22]Livak KJ, Schmittgen TD. Analysis of relative gene expression data using real-time quantitative PCR and the 2(−Delta Delta C(T)) Method. Methods. 2001; 25(4):402-8.
• [23]Tavazoie SF, Alarcon C, Oskarsson T, Padua D, Wang Q, Bos PD et al.. Endogenous human microRNAs that suppress breast cancer metastasis. Nature. 2008; 451(7175):147-52.
• [24]Guil S, Gattoni R, Carrascal M, Abián J, Stévenin J, Bach-Elias M. Roles of hnRNP A1, SR proteins, and p68 Helicase in c-H-ras Alternative Splicing Regulation. Mol Cell Biol. 2003; 23(8):2927-41.
• [25]Aoki Y, Niihori T, Kawame H, Kurosawa K, Ohashi H, Tanaka Y et al.. Germline mutations in HRAS proto-oncogene cause Costello syndrome. Nat Genet. 2005; 37(10):1038-40.
• [26]Miko E, Czimmerer Z, Csanky E, Boros G, Buslig J, Dezso B et al.. Differentially expressed microRNAs in small cell lung cancer. Exp Lung Res. 2009; 35(8):646-64.
• [27]Bussing I, Slack FJ, Grosshans H. let-7 microRNAs in development, stem cells and cancer. Trends Mol Med. 2008; 14(9):400-9.
• [28]Esteban LM, Vicario-Abejon C, Fernandez-Salguero P, Fernandez-Medarde A, Swaminathan N, Yienger K et al.. Targeted genomic disruption of H-ras and N-ras, individually or in combination, reveals the dispensability of both loci for mouse growth and development. Mol Cell Biol. 2001; 21(5):1444-52.
• [29]Chiu SC, Chung HY, Cho DY, Chan TM, Liu MC, Huang HM et al.. Therapeutic potential of microRNA let-7: tumor suppression or impeding normal stemness. Cell Transplant. 2014; 23(4–5):459-69.
• [30]Garzon R, Pichiorri F, Palumbo T, Visentini M, Aqeilan R, Cimmino A et al. MicroRNA gene expression during retinoic acid-induced differentiation of human acute promyelocytic leukemia. Oncogene. 2007.
• [31]Mosakhani N, Räty R, Tyybakinoja A, Karjalainen-Lindsberg M, Elonen ET, Knuutila S. MicroRNA Profiling in Chemoresistant and Chemosensitive Acute. Myeloid Leukemia. Cytogenet Genome Res. 2013;141(4):272-6.
• [32]Adams BD, Furneaux H, White BA. The micro-ribonucleic acid (miRNA) miR-206 targets the human estrogen receptor-alpha (ERalpha) and represses ERalpha messenger RNA and protein expression in breast cancer cell lines. Mol Endocrinol. 2007; 21(5):1132-47.
• [33]Gambardella S, Rinaldi F, Lepore SM, Viola A, Loro E, Angelini C et al.. Overexpression of microRNA-206 in the skeletal muscle from myotonic dystrophy type 1 patients. J Transl Med. 2010; 8(1):48. BioMed Central Full Text
• [34]Ma G, Wang Y, Li Y, Cui L, Zhao Y, Zhao B et al.. MiR-206, a key modulator of skeletal muscle development and disease. Int J Biol Sci. 2015; 11(3):345-52.
• [35]Vimalraj S, Miranda PJ, Ramyakrishna B, Selvamurugan N. Regulation of breast cancer and bone metastasis by microRNAs. Dis Markers. 2013; 35(5):369-87.
• [36]Bicker S, Lackinger M, Weiss K, Schratt G. MicroRNA-132, −134, and −138: a microRNA troika rules in neuronal dendrites. Cell Mol Life Sci. 2014; 71(20):3987-4005.
• [37]Chen X, Yan Q, Li S, Zhou L, Yang H, Yang Y et al.. Expression of the tumor suppressor miR-206 is associated with cellular proliferative inhibition and impairs invasion in ERalpha-positive endometrioid adenocarcinoma. Cancer Lett. 2012; 314(1):41-53.
• [38]Dereure O. Mutations in H-Ras proto-oncogen in Costello syndrome. Ann Dermatol Venereol. 2006; 133(8–9 Pt 1):731.
• [39]Paquin A, Hordo C, Kaplan DR, Miller FD. Costello syndrome H-Ras alleles regulate cortical development. Dev Biol. 2009; 330(2):440-51.