Frequent downregulation of LRRC26 by epigenetic alterations is involved in the malignant progression of triple-negative breast cancer

Miyagawa,Yoshimasa; Matsushita,Yosuke; Suzuki,Hiromu; Komatsu,Masato; Yoshimaru,Tetsuro; Kimura,Ryuichiro; Yanai,Ayako; Honda,Junko; Tangoku,Akira; Sasa,Mitsunori; Miyoshi,Yasuo; Katagiri,Toyomasa

doi:10.3892/ijo.2018.4301

Frequent downregulation of LRRC26 by epigenetic alterations is involved in the malignant progression of triple-negative breast cancer

Authors:
- Yoshimasa Miyagawa
- Yosuke Matsushita
- Hiromu Suzuki
- Masato Komatsu
- Tetsuro Yoshimaru
- Ryuichiro Kimura
- Ayako Yanai
- Junko Honda
- Akira Tangoku
- Mitsunori Sasa
- Yasuo Miyoshi
- Toyomasa Katagiri
View Affiliations

Affiliations: Division of Genome Medicine, Institute for Genome Research, Tokushima University, Tokushima 770-8503, Japan, Department of Molecular Biology, Sapporo Medical University, Sapporo, Hokkaido 060-8556, Japan, Department of Surgery, National Hospital Organization Higashitokushima Medical Center, Tokushima 779-0193, Japan, Department of Thoracic and Endocrine Surgery and Oncology, Institute of Health Biosciences, The University of Tokushima Graduate School, Tokushima 770-8503, Japan, Department of Surgery, Tokushima Breast Care Clinic, Tokushima 770-0052, Japan, Department of Surgery, Division of Breast and Endocrine Surgery, Hyogo College of Medicine, Nishinomiya, Hyogo 663-8501, Japan
Published online on: March 5, 2018 https://doi.org/10.3892/ijo.2018.4301
Pages: 1539-1558

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

Triple-negative breast cancer (TNBC), defined as breast cancer lacking estrogen- and progesterone‑receptor expression and human epidermal growth factor receptor 2 (HER2) amplification, is a heterogeneous disease. RNA-sequencing analysis of 15 TNBC specimens and The Cancer Genome Atlas-TNBC dataset analysis identified the frequent downregulation of leucine-rich repeat-containing 26 (LRRC26), which negatively regulates nuclear factor-κB (NF-κB) signaling, in TNBC tissues. Quantitative polymerase chain reaction and bisulfite pyrosequencing analyses revealed that LRRC26 was frequently silenced in TNBC tissues and cell lines as a result of promoter methylation. LRRC26 expression was restored by 5-aza-2'-deoxycytidine (5'-aza-dC) treatment in HCC1937 TNBC cells, which lack LRRC26 expression. Notably, small interfering RNA-mediated knockdown of LRRC26 expression significantly enhanced the anchorage-independent growth, invasion and migration of HCC70 cells, whereas ectopic overexpression of LRRC26 in BT20 cells suppressed their invasion and migration. Conversely, neither knockdown nor overexpression of LRRC26 had an effect on cell viability in the absence of tumor necrosis factor-α (TNF-α) stimulation. Meanwhile, overexpression of LRRC26 caused the reduction of TNF-α-mediated NF-κB luciferase reporter activity, whereas depleting LRRC26 expression resulted in the upregulation of TNF-α-mediated NF-κB downstream genes [interleukin-6 (IL-6), IL-8 and C-X-C motif chemokine ligand-1]. Taken together, these findings demonstrate that LRRC26 is frequently downregulated in TNBC due to DNA methylation and that it suppresses the TNF-α-independent anchorage-independent growth, invasion and migration of TNBC cells. Loss of LRRC26 function may be a critical event in the aggressiveness of TNBC cells through a TNF-α/NF-κB-independent mechanism.

View Figures

View References

1	Fisher B, Anderson S, Tan-Chiu E, Wolmark N, Wickerham DL, Fisher ER, Dimitrov NV, Atkins JN, Abramson N, Merajver S, et al: Tamoxifen and chemotherapy for axillary node-negative, estrogen receptor-negative breast cancer: Findings from National Surgical Adjuvant Breast and Bowel Project B-23. J Clin Oncol. 19:931–942. 2001. View Article : Google Scholar : PubMed/NCBI
2	Colleoni M, Gelber S, Goldhirsch A, Aebi S, Castiglione-Gertsch M, Price KN, Coates AS and Gelber RD; International Breast Cancer Study Group: Tamoxifen after adjuvant chemotherapy for premenopausal women with lymph node-positive breast cancer: International Breast Cancer Study Group Trial 13-93. J Clin Oncol. 24:1332–1341. 2006. View Article : Google Scholar : PubMed/NCBI
3	Merglen A, Verkooijen HM, Fioretta G, Neyroud-Caspar I, Vinh-Hung V, Vlastos G, Chappuis PO, Castiglione M, Rapiti E and Bouchardy C: Hormonal therapy for oestrogen receptor-negative breast cancer is associated with higher disease-specific mortality. Ann Oncol. 20:857–861. 2009. View Article : Google Scholar : PubMed/NCBI
4	Huszno J and Nowara E: Current therapeutic strategies of anti-HER2 treatment in advanced breast cancer patients. Contemp Oncol (Pozn). 20:1–7. 2016.
5	Foulkes WD, Smith IE and Reis-Filho JS: Triple-negative breast cancer. N Engl J Med. 363:1938–1948. 2010. View Article : Google Scholar : PubMed/NCBI
6	Liedtke C, Mazouni C, Hess KR, André F, Tordai A, Mejia JA, Symmans WF, Gonzalez-Angulo AM, Hennessy B, Green M, et al: Response to neoadjuvant therapy and long-term survival in patients with triple-negative breast cancer. J Clin Oncol. 26:1275–1281. 2008. View Article : Google Scholar : PubMed/NCBI
7	Gradishar WJ, Anderson BO, Balassanian R, Blair SL, Burstein HJ, Cyr A, Elias AD, Farrar WB, Forero A, Giordano SH, et al: Invasive Breast Cancer Version 1.2016, NCCN Clinical Practice Guidelines in Oncology. J Natl Compr Canc Netw. 14:324–354. 2016. View Article : Google Scholar
8	Cortes J, O'Shaughnessy J, Loesch D, Blum JL, Vahdat LT, Petrakova K, Chollet P, Manikas A, Diéras V, Delozier T, et al EMBRACE (Eisai Metastatic Breast Cancer Study Assessing Physician's Choice Versus E7389) investigators: Eribulin mono-therapy versus treatment of physician's choice in patients with metastatic breast cancer (EMBRACE): A phase 3 open-label randomised study. Lancet. 377:914–923. 2011. View Article : Google Scholar : PubMed/NCBI
9	Denkert C, Liedtke C, Tutt A and von Minckwitz G: Molecular alterations in triple-negative breast cancer-the road to new treatment strategies. Lancet. 389:2430–2442. 2017. View Article : Google Scholar
10	Kim JE, Ahn HJ, Ahn JH, Yoon DH, Kim SB, Jung KH, Gong GY, Kim MJ, Son BH and Ahn SH: Impact of triple-negative breast cancer phenotype on prognosis in patients with stage I breast cancer. J Breast Cancer. 15:197–202. 2012. View Article : Google Scholar : PubMed/NCBI
11	Lehmann BD, Bauer JA, Chen X, Sanders ME, Chakravarthy AB, Shyr Y and Pietenpol JA: Identification of human triple-negative breast cancer subtypes and preclinical models for selection of targeted therapies. J Clin Invest. 121:2750–2767. 2011. View Article : Google Scholar : PubMed/NCBI
12	Cancer Genome Atlas N; Cancer Genome Atlas Network: Comprehensive molecular portraits of human breast tumours. Nature. 490:61–70. 2012. View Article : Google Scholar : PubMed/NCBI
13	Pereira B, Chin SF, Rueda OM, Vollan HK, Provenzano E, Bardwell HA, Pugh M, Jones L, Russell R, Sammut SJ, et al: The somatic mutation profiles of 2,433 breast cancers refines their genomic and transcriptomic landscapes. Nat Commun. 7:114792016. View Article : Google Scholar : PubMed/NCBI
14	Shah SP, Roth A, Goya R, Oloumi A, Ha G, Zhao Y, Turashvili G, Ding J, Tse K, Haffari G, et al: The clonal and mutational evolution spectrum of primary triple-negative breast cancers. Nature. 486:395–399. 2012.PubMed/NCBI
15	O'Shaughnessy J, Osborne C, Pippen JE, Yoffe M, Patt D, Rocha C, Koo IC, Sherman BM and Bradley C: Iniparib plus chemotherapy in metastatic triple-negative breast cancer. N Engl J Med. 364:205–214. 2011. View Article : Google Scholar : PubMed/NCBI
16	Tutt A, Robson M, Garber JE, Domchek SM, Audeh MW, Weitzel JN, Friedlander M, Arun B, Loman N, Schmutzler RK, et al: Oral poly(ADP-ribose) polymerase inhibitor olaparib in patients with BRCA1 or BRCA2 mutations and advanced breast cancer: A proof-of-concept trial. Lancet. 376:235–244. 2010. View Article : Google Scholar : PubMed/NCBI
17	Yamaguchi N, Ito T, Azuma S, Ito E, Honma R, Yanagisawa Y, Nishikawa A, Kawamura M, Imai J, Watanabe S, et al: Constitutive activation of nuclear factor-kappaB is preferentially involved in the proliferation of basal-like subtype breast cancer cell lines. Cancer Sci. 100:1668–1674. 2009. View Article : Google Scholar : PubMed/NCBI
18	Liu XF, Xiang L, Zhang Y, Becker KG, Bera TK and Pastan I: CAPC negatively regulates NF-κB activation and suppresses tumor growth and metastasis. Oncogene. 31:1673–1682. 2012. View Article : Google Scholar
19	Kurebayashi J, Kurosumi M and Sonoo H: A new human breast cancer cell line, KPL-3C, secretes parathyroid hormone-related protein and produces tumours associated with microcalcifications in nude mice. Br J Cancer. 74:200–207. 1996. View Article : Google Scholar : PubMed/NCBI
20	Komatsu M, Yoshimaru T, Matsuo T, Kiyotani K, Miyoshi Y, Tanahashi T, Rokutan K, Yamaguchi R, Saito A, Imoto S, et al: Molecular features of triple negative breast cancer cells by genome-wide gene expression profiling analysis. Int J Oncol. 42:478–506. 2013. View Article : Google Scholar
21	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
22	Kim JW, Akiyama M, Park JH, Lin ML, Shimo A, Ueki T, Daigo Y, Tsunoda T, Nishidate T, Nakamura Y, et al: Activation of an estrogen/estrogen receptor signaling by BIG3 through its inhibitory effect on nuclear transport of PHB2/REA in breast cancer. Cancer Sci. 100:1468–1478. 2009. View Article : Google Scholar : PubMed/NCBI
23	Park JH, Lin ML, Nishidate T, Nakamura Y and Katagiri T: PDZ-binding kinase/T-LAK cell-originated protein kinase, a putative cancer/testis antigen with an oncogenic activity in breast cancer. Cancer Res. 66:9186–9195. 2006. View Article : Google Scholar : PubMed/NCBI
24	Kamimae S, Yamamoto E, Kai M, Niinuma T, Yamano HO, Nojima M, Yoshikawa K, Kimura T, Takagi R, Harada E, et al: Epigenetic silencing of NTSR1 is associated with lateral and noninvasive growth of colorectal tumors. Oncotarget. 6:29975–29990. 2015. View Article : Google Scholar : PubMed/NCBI
25	Ng FW, Nguyen M, Kwan T, Branton PE, Nicholson DW, Cromlish JA and Shore GC: p28 Bap31, a Bcl-2/Bcl-XL- and procaspase-8-associated protein in the endoplasmic reticulum. J Cell Biol. 139:327–338. 1997. View Article : Google Scholar : PubMed/NCBI
26	Annaert WG, Becker B, Kistner U, Reth M and Jahn R: Export of cellubrevin from the endoplasmic reticulum is controlled by BAP31. J Cell Biol. 139:1397–1410. 1997. View Article : Google Scholar
27	Huang W, Sherman BT and Lempicki RA: Systematic and integrative analysis of large gene lists using DAVID bioinformatics resources. Nat Protoc. 4:44–57. 2009. View Article : Google Scholar
28	Huang W, Sherman BT and Lempicki RA: Bioinformatics enrichment tools: Paths toward the comprehensive functional analysis of large gene lists. Nucleic Acids Res. 37:1–13. 2009. View Article : Google Scholar :
29	Hao JM, Chen JZ, Sui HM, Si-Ma XQ, Li GQ, Liu C, Li JL, Ding YQ and Li JM: A five-gene signature as a potential predictor of metastasis and survival in colorectal cancer. J Pathol. 220:475–489. 2010.PubMed/NCBI
30	Yoshimaru T, Komatsu M, Matsuo T, Chen YA, Murakami Y, Mizuguchi K, Mizohata E, Inoue T, Akiyama M, Yamaguchi R, et al: Targeting BIG3-PHB2 interaction to overcome tamoxifen resistance in breast cancer cells. Nat Commun. 4:24432013. View Article : Google Scholar : PubMed/NCBI
31	Kajiura K, Masuda K, Naruto T, Kohmoto T, Watabnabe M, Tsuboi M, Takizawa H, Kondo K, Tangoku A and Imoto I: Frequent silencing of the candidate tumor suppressor TRIM58 by promoter methylation in early-stage lung adenocarcinoma. Oncotarget. 8:2890–2905. 2017. View Article : Google Scholar :
32	Locke WJ and Clark SJ: Epigenome remodelling in breast cancer: Insights from an early in vitro model of carcinogenesis. Breast Cancer Res. 14:2152012. View Article : Google Scholar : PubMed/NCBI
33	Wang X: miRDB: A microRNA target prediction and functional annotation database with a wiki interface. RNA. 14:1012–1017. 2008. View Article : Google Scholar : PubMed/NCBI
34	Peña-Chilet M, Martínez MT, Pérez-Fidalgo JA, Peiró-Chova L, Oltra SS, Tormo E, Alonso-Yuste E, Martinez-Delgado B, Eroles P, Climent J, et al: MicroRNA profile in very young women with breast cancer. BMC Cancer. 14:5292014. View Article : Google Scholar : PubMed/NCBI
35	Nguyen HT, Li C, Lin Z, Zhuang Y, Flemington EK, Burow ME, Lin YI and Shan B: The microRNA expression associated with morphogenesis of breast cancer cells in three-dimensional organotypic culture. Oncol Rep. 28:117–126. 2012.PubMed/NCBI
36	Yan J and Aldrich RW: BK potassium channel modulation by leucine-rich repeat-containing proteins. Proc Natl Acad Sci USA. 109:7917–7922. 2012. View Article : Google Scholar : PubMed/NCBI
37	Carvalho S, Oliveira T, Bartels MF, Miyoshi E, Pierce M, Taniguchi N, Carneiro F, Seruca R, Reis CA, Strahl S, et al: O-mannosylation and N-glycosylation: Two coordinated mechanisms regulating the tumour suppressor functions of E-cadherin in cancer. Oncotarget. 7:65231–65246. 2016. View Article : Google Scholar : PubMed/NCBI
38	Couldrey C and Green JE: Metastases: The glycan connection. Breast Cancer Res. 2:321–323. 2000. View Article : Google Scholar
39	Dall'Olio F, Malagolini N, Trinchera M and Chiricolo M: Mechanisms of cancer-associated glycosylation changes. Front Biosci (Landmark Ed). 17:670–699. 2012. View Article : Google Scholar
40	Pinho SS and Reis CA: Glycosylation in cancer: Mechanisms and clinical implications. Nat Rev Cancer. 15:540–555. 2015. View Article : Google Scholar : PubMed/NCBI
41	Wolff AC, Hammond ME, Hicks DG, Dowsett M, McShane LM, Allison KH, Allred DC, Bartlett JM, Bilous M, Fitzgibbons P, et al American Society of Clinical Oncology; College of American Pathologists: Recommendations for human epidermal growth factor receptor 2 testing in breast cancer: American Society of Clinical Oncology/College of American Pathologists clinical practice guideline update. J Clin Oncol. 31:3997–4013. 2013. View Article : Google Scholar : PubMed/NCBI
42	Sobin L, Gospodarowicz M and Wittekind C: TNM Classification of Malignant Tumors. John Wiley & Sons, Inc; Hoboken, NJ: 2010