The long non-coding RNA GHSROS facilitates breast cancer cell migration and orthotopic xenograft tumour growth

Thomas,Patrick B.; Seim,Inge; Jeffery,Penny L.; Gahete,Manuel D.; Maugham,Michelle; Crisp,Gabrielle J.; Stacey,Andrew; Shah,Esha T.; Walpole,Carina; Whiteside,Eliza J.; Nelson,Colleen C.; Herington,Adrian C.; Luque,Raúl M.; Veedu,Rakesh N.; Chopin,Lisa K.

doi:10.3892/ijo.2019.4891

The long non-coding RNA GHSROS facilitates breast cancer cell migration and orthotopic xenograft tumour growth

Authors:
- Patrick B. Thomas
- Inge Seim
- Penny L. Jeffery
- Manuel D. Gahete
- Michelle Maugham
- Gabrielle J. Crisp
- Andrew Stacey
- Esha T. Shah
- Carina Walpole
- Eliza J. Whiteside
- Colleen C. Nelson
- Adrian C. Herington
- Raúl M. Luque
- Rakesh N. Veedu
- Lisa K. Chopin
View Affiliations

Affiliations: Ghrelin Research Group, Translational Research Institute-Institute of Health and Biomedical Innovation, School of Biomedical Sciences, Queensland University of Technology, Brisbane, Queensland 4102, Australia, Maimonides Institute of Biomedical Research of Cordoba (IMIBIC), 14004 Córdoba, Spain, Moffitt Cancer Center and Research Institute, Tampa, FL, 33612, USA, Australian Prostate Cancer Research Centre-Queensland, Translational Research Institute, Brisbane, Queensland 4102, Australia, Centre for Comparative Genomics, Murdoch University & Perron Institute for Neurological and Translational Science, Perth, Western Australia 6150, Australia
Published online on: October 4, 2019 https://doi.org/10.3892/ijo.2019.4891
Pages: 1223-1236
Copyright: © Thomas et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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

Recent evidence suggests that numerous long non‑coding RNAs (lncRNAs) are dysregulated in cancer, and have critical roles in tumour development and progression. The present study investigated the ghrelin receptor antisense lncRNA growth hormone secretagogue receptor opposite strand (GHSROS) in breast cancer. Reverse transcription‑quantitative polymerase chain reaction revealed that GHSROS expression was significantly upregulated in breast tumour tissues compared with normal breast tissue. Induced overexpression of GHSROS in the MDA‑MB‑231 breast cancer cell line significantly increased cell migration in vitro, without affecting cell proliferation, a finding similar to our previous study on lung cancer cell lines. Microarray analysis revealed a significant repression of a small cluster of major histocompatibility class II genes and enrichment of immune response pathways; this phenomenon may allow tumour cells to better evade the immune system. Ectopic overexpression of GHSROS in the MDA‑MB‑231 cell line significantly increased orthotopic xenograft growth in mice, suggesting that in vitro culture does not fully capture the function of this lncRNA. This study demonstrated that GHSROS may serve a relevant role in breast cancer. Further studies are warranted to explore the function and therapeutic potential of this lncRNA in breast cancer progression.

View Figures

View References

1	Mattick JS and Rinn JL: Discovery and annotation of long noncoding RNAs. Nat Struct Mol Biol. 22:5–7. 2015. View Article : Google Scholar : PubMed/NCBI
2	Deveson IW, Hardwick SA, Mercer TR and Mattick JS: The Dimensions, dynamics, and relevance of the mammalian noncoding transcriptome. Trends Genet. 33:464–478. 2017. View Article : Google Scholar : PubMed/NCBI
3	Hu X, Sood AK, Dang CV and Zhang L: The role of long noncoding RNAs in cancer: The dark matter matters. Curr Opin Genet Dev. 48:8–15. 2018. View Article : Google Scholar :
4	Quinn JJ and Chang HY: Unique features of long non-coding RNA biogenesis and function. Nat Rev Genet. 17:47–62. 2016. View Article : Google Scholar
5	Sun M and Kraus WL: From discovery to function: The expanding roles of long non-coding RNAs in physiology and disease. Endocr Rev. Jan 7–2015.Epub ahead of print. View Article : Google Scholar
6	Jemal A, Bray F, Center MM, Ferlay J, Ward E and Forman D: Global cancer statistics. CA Cancer J Clin. 61:69–90. 2011. View Article : Google Scholar : PubMed/NCBI
7	Australian Institute of Health and Welfare: Cancer in Australia: Actual incidence data from 1982 to 2013 and mortality data from 1982 to 2014 with projections to 2017. Asia Pac J Clin Oncol. 14:5–15. 2018. View Article : Google Scholar
8	ENCODE Project Consortium: The ENCODE (ENCyclopedia Of DNA Elements) Project. Science. 306:636–640. 2004. View Article : Google Scholar : PubMed/NCBI
9	Kumar M, DeVaux RS and Herschkowitz JI: Molecular and cellular changes in breast cancer and new roles of lncRNAs in breast cancer initiation and progression. Prog Mol Biol Transl Sci. 144:563–586. 2016. View Article : Google Scholar : PubMed/NCBI
10	Amorim M, Salta S, Henrique R and Jerónimo C: Decoding the usefulness of non-coding RNAs as breast cancer markers. J Transl Med. 14:2652016. View Article : Google Scholar : PubMed/NCBI
11	Cerk S, Schwarzenbacher D, Adiprasito JB, Stotz M, Hutterer GC, Gerger A, Ling H, Calin GA and Pichler M: Current status of long non-coding RNAs in human breast cancer. Int J Mol Sci. 17:172016. View Article : Google Scholar
12	Soudyab M, Iranpour M and Ghafouri-Fard S: The role of long non-coding RNAs in breast cancer. Arch Iran Med. 19:508–517. 2016.PubMed/NCBI
13	Bhan A, Hussain I, Ansari KI, Kasiri S, Bashyal A and Mandal SS: Antisense transcript long noncoding RNA (lncRNA) HOTAIR is transcriptionally induced by estradiol. J Mol Biol. 425:3707–3722. 2013. View Article : Google Scholar : PubMed/NCBI
14	He X, Bao W, Li X, Chen Z, Che Q, Wang H and Wan XP: The long non-coding RNA HOTAIR is upregulated in endometrial carcinoma and correlates with poor prognosis. Int J Mol Med. 33:325–332. 2014. View Article : Google Scholar
15	Bhan A and Mandal SS: Estradiol-induced transcriptional regulation of long non-coding RNA, HOTAIR. Methods Mol Biol. 1366:395–412. 2016. View Article : Google Scholar
16	Xue X, Yang YA, Zhang A, Fong KW, Kim J, Song B, Li S, Zhao JC and Yu J: LncRNA HOTAIR enhances ER signaling and confers tamoxifen resistance in breast cancer. Oncogene. 35:2746–2755. 2016. View Article : Google Scholar :
17	Aiello A, Bacci L, Re A, Ripoli C, Pierconti F, Pinto F, Masetti R, Grassi C, Gaetano C, Bassi PF, et al: MALAT1 and HOTAIR long non-coding RNAs play opposite role in estrogen-mediated transcriptional regulation in prostate cancer cells. Sci Rep. 6:384142016. View Article : Google Scholar : PubMed/NCBI
18	Arun G, Diermeier S, Akerman M, Chang KC, Wilkinson JE, Hearn S, Kim Y, MacLeod AR, Krainer AR, Norton L, et al: Differentiation of mammary tumors and reduction in metastasis upon Malat1 lncRNA loss. Genes Dev. 30:34–51. 2016. View Article : Google Scholar :
19	Whiteside EJ, Seim I, Pauli JP, O'Keeffe AJ, Thomas PB, Carter SL, Walpole CM, Fung JN, Josh P, Herington AC, et al: Identification of a long non-coding RNA gene, growth hormone secretagogue receptor opposite strand, which stimulates cell migration in non-small cell lung cancer cell lines. Int J Oncol. 43:566–574. 2013. View Article : Google Scholar : PubMed/NCBI
20	Livak KJ and Schmittgen TD: Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) method. Methods. 25:402–408. 2001. View Article : Google Scholar
21	Ke N, Wang X, Xu X and Abassi YA: The xCELLigence system for real-time and label-free monitoring of cell viability. Methods Mol Biol. 740:33–43. 2011. View Article : Google Scholar : PubMed/NCBI
22	Limame R, Wouters A, Pauwels B, Fransen E, Peeters M, Lardon F, De Wever O and Pauwels P: Comparative analysis of dynamic cell viability, migration and invasion assessments by novel real-time technology and classic endpoint assays. PLoS One. 7:e465362012. View Article : Google Scholar : PubMed/NCBI
23	Ritchie ME, Phipson B, Wu D, Hu Y, Law CW, Shi W and Smyth GK: limma powers differential expression analyses for RNA-sequencing and microarray studies. Nucleic Acids Res. 43:e472015. View Article : Google Scholar : PubMed/NCBI
24	Szklarczyk D, Morris JH, Cook H, Kuhn M, Wyder S, Simonovic M, Santos A, Doncheva NT, Roth A, Bork P, et al: The STRING database in 2017: Quality-controlled protein-protein association networks, made broadly accessible. Nucleic Acids Res. 45:D362–D368. 2017. View Article : Google Scholar
25	Hartigan JA and Wong MA: Algorithm AS 136: A k-means clustering algorithm. J R Stat Soc Ser C Appl Stat. 28:100–108. 1979.
26	Du J, Yuan Z, Ma Z, Song J, Xie X and Chen Y: KEGG-PATH: Kyoto encyclopedia of genes and genomes-based pathway analysis using a path analysis model. Mol Biosyst. 10:2441–2447. 2014. View Article : Google Scholar : PubMed/NCBI
27	Russell PJ, Raghavan D, Gregory P, Philips J, Wills EJ, Jelbart M, Wass J, Zbroja RA and Vincent PC: Bladder cancer xenografts: A model of tumor cell heterogeneity. Cancer Res. 46:2035–2040. 1986.PubMed/NCBI
28	Lim E, Modi KD and Kim J: In vivo bioluminescent imaging of mammary tumors using IVIS spectrum. J Vis Exp. Apr 29–2009.Epub ahead of print. View Article : Google Scholar
29	Amir S, Simion C, Umeh-Garcia M, Krig S, Moss T, Carraway KL III and Sweeney C: Regulation of the T-box transcription factor Tbx3 by the tumour suppressor microRNA-206 in breast cancer. Br J Cancer. 114:1125–1134. 2016. View Article : Google Scholar : PubMed/NCBI
30	Trowsdale J and Knight JC: Major histocompatibility complex genomics and human disease. Annu Rev Genomics Hum Genet. 14:301–323. 2013. View Article : Google Scholar : PubMed/NCBI
31	Boegel S, Löwer M, Schäfer M, Bukur T, de Graaf J, Boisguérin V, Türeci O, Diken M, Castle JC and Sahin U: HLA typing from RNA-Seq sequence reads. Genome Med. 4:1022012. View Article : Google Scholar : PubMed/NCBI
32	Tabibzadeh SS, Sivarajah A, Carpenter D, Ohlsson-Wilhelm BM and Satyaswaroop PG: Modulation of HLA-DR expression in epithelial cells by interleukin 1 and estradiol-17 beta. J Clin Endocrinol Metab. 71:740–747. 1990. View Article : Google Scholar : PubMed/NCBI
33	Kocatürk B and Versteeg HH: Orthotopic injection of breast cancer cells into the mammary fat pad of mice to study tumor growth. J Vis Exp. Feb 8–2015.Epub ahead of print. View Article : Google Scholar : PubMed/NCBI
34	Lanzós A, Carlevaro-Fita J, Mularoni L, Reverter F, Palumbo E, Guigó R and Johnson R: Discovery of cancer driver long noncoding RNAs across 1112 tumour genomes: New candidates and distinguishing features. Sci Rep. 7:415442017. View Article : Google Scholar : PubMed/NCBI
35	Qiu MT, Hu JW, Yin R and Xu L: Long noncoding RNA: An emerging paradigm of cancer research. Tumour Biol. 34:613–620. 2013. View Article : Google Scholar : PubMed/NCBI
36	Wenric S, ElGuendi S, Caberg JH, Bezzaou W, Fasquelle C, Charloteaux B, Karim L, Hennuy B, Frères P, Collignon J, et al: Transcriptome-wide analysis of natural antisense transcripts shows their potential role in breast cancer. Sci Rep. 7:174522017. View Article : Google Scholar : PubMed/NCBI
37	Nie L, Wu HJ, Hsu JM, Chang SS, Labaff AM, Li CW, Wang Y, Hsu JL and Hung MC: Long non-coding RNAs: Versatile master regulators of gene expression and crucial players in cancer. Am J Transl Res. 4:127–150. 2012.PubMed/NCBI
38	Pan YF, Feng L, Zhang XQ, Song LJ, Liang HX, Li ZQ and Tao FB: Role of long non-coding RNAs in gene regulation and oncogenesis. Chin Med J (Engl). 124:2378–2383. 2011.
39	Collette J, Le Bourhis X and Adriaenssens E: Regulation of human breast cancer by the long non-coding RNA H19. Int J Mol Sci. 18:182017. View Article : Google Scholar
40	Miao Y, Fan R, Chen L and Qian H: Clinical significance of long non-coding RNA MALAT1 expression in tissue and serum of breast cancer. Ann Clin Lab Sci. 46:418–424. 2016.PubMed/NCBI
41	Avazpour N, Hajjari M and Tahmasebi Birgani M: HOTAIR: A promising long non-coding RNA with potential role in breast invasive carcinoma. Front Genet. 8:1702017. View Article : Google Scholar :
42	Chiu HS, Somvanshi S, Patel E, Chen TW, Singh VP, Zorman B, Patil SL, Pan Y, Chatterjee SS, Sood AK, et al Cancer Genome Atlas Research Network: Pan-Cancer Analysis of lncRNA regulation supports their targeting of cancer genes in each tumor context. Cell Rep. 23:297–312.e12. 2018. View Article : Google Scholar : PubMed/NCBI
43	Iorns E, Drews-Elger K, Ward TM, Dean S, Clarke J, Berry D, El Ashry D and Lippman M: A new mouse model for the study of human breast cancer metastasis. PLoS One. 7:e479952012. View Article : Google Scholar : PubMed/NCBI
44	Nagini S: Breast Cancer: Current molecular therapeutic targets and new players. Anticancer Agents Med Chem. 17:152–163. 2017. View Article : Google Scholar
45	Cavo M, Fato M, Peñuela L, Beltrame F, Raiteri R and Scaglione S: Microenvironment complexity and matrix stiffness regulate breast cancer cell activity in a 3D in vitro model. Sci Rep. 6:353672016. View Article : Google Scholar : PubMed/NCBI
46	Kopparapu PK, Tinzl M, Anagnostaki L, Persson JL and Dizeyi N: Expression and localization of serotonin receptors in human breast cancer. Anticancer Res. 33:363–370. 2013.PubMed/NCBI
47	Pai VP, Marshall AM, Hernandez LL, Buckley AR and Horseman ND: Altered serotonin physiology in human breast cancers favors paradoxical growth and cell survival. Breast Cancer Res. 11:R812009. View Article : Google Scholar : PubMed/NCBI
48	Li J, Weinberg MS, Zerbini L and Prince S: The oncogenic TBX3 is a downstream target and mediator of the TGF-β1 signaling pathway. Mol Biol Cell. 24:3569–3576. 2013. View Article : Google Scholar : PubMed/NCBI
49	Peres J, Davis E, Mowla S, Bennett DC, Li JA, Wansleben S and Prince S: The highly homologous T-Box transcription factors, TBX2 and TBX3, have distinct roles in the oncogenic process. Genes Cancer. 1:272–282. 2010. View Article : Google Scholar : PubMed/NCBI
50	Forero A, Li Y, Chen D, Grizzle WE, Updike KL, Merz ND, Downs-Kelly E, Burwell TC, Vaklavas C, Buchsbaum DJ, et al: Expression of the MHC Class II pathway in triple-negative breast cancer tumor cells is associated with a good prognosis and infiltrating lymphocytes. Cancer Immunol Res. 4:390–399. 2016. View Article : Google Scholar : PubMed/NCBI
51	Doonan BP and Haque A: HLA Class II antigen presentation in prostate cancer cells: A novel approach to prostate tumor immu-notherapy. Open Cancer Immunol J. 3:1–7. 2010. View Article : Google Scholar : PubMed/NCBI
52	Thibodeau J, Bourgeois-Daigneault MC and Lapointe R: Targeting the MHC Class II antigen presentation pathway in cancer immunotherapy. OncoImmunology. 1:908–916. 2012. View Article : Google Scholar : PubMed/NCBI
53	Walsh NC, Kenney LL, Jangalwe S, Aryee KE, Greiner DL, Brehm MA and Shultz LD: Humanized mouse models of clinical disease. Annu Rev Pathol. 12:187–215. 2017. View Article : Google Scholar :
54	Zhao Y, Shuen TWH, Toh TB, Chan XY, Liu M, Tan SY, Fan Y, Yang H, Lyer SG, Bonney GK, et al: Development of a new patient-derived xenograft humanised mouse model to study human-specific tumour microenvironment and immunotherapy. Gut. 67:1845–1854. 2018. View Article : Google Scholar : PubMed/NCBI