miR-342-5p as a Potential Regulator of HER2 Breast Cancer Cell Growth

Author(s): Evita Maria Lindholm*, Suvi-Katri Leivonen, Eldri Undlien, Daniel Nebdal, Anna Git, Carlos Caldas, Anne-Lise Børresen-Dale, Kristine Kleivi

Journal Name: MicroRNA

Volume 8 , Issue 2 , 2019

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Graphical Abstract:


Background: HER2 positive Breast Cancers (BC) have aggressive behavior and poor prognosis. Previously, we have identified miR-342-5p as an upstream regulator of HER2 signaling, as well as inhibitor of HER2 positive BC cell line growth.

Objective: Here, we aimed to further investigate the molecular mechanisms behind miR-342-5pinduced HER2 pathway deregulation.

Method: Two HER2 amplified breast cancer cell lines were transiently transfected with miR-342-5p mimic or negative control, and gene expression was analyzed by Agilent microarrays. Three clinical datasets with BC patients were used to identify correlations between candidate genes and miR-342- 5p, and associations with survival.

Results: Pathway analyses of all deregulated genes revealed a significant suppression of the HER2 downstream pathways ERK/MAPK and SAPK/JNK, whereas the miR-342-5p predicted target genes were enriched for pathways associated with cell motility.Biological functions linked to mitochondrial stability were ranked among the top toxicological functions in both gene lists. Among the most deregulated genes, Cytochrome B5 Reductase 3 (CYB5R3) and Rap Guanine Nucleotide Exchange Factor 6 (RAPGEF6) significantly anticorrelated and correlated, respectively, with miR-342-5p in all three clinical BC datasets. Low CYB5R3 levels and high RAPGEF6 levels were significantly associated with survival, although this was not directly associated with HER2 expression.

Conclusion: Our data suggest that miR-342-5p overexpression in HER2 positive BC cell lines elicits broad effects on HER2 downstream signaling, cell motility and mitochondrial stability. Together these effects may render cells less proliferative and more sensitive to cellular stress.

Keywords: CYB5R3, HER2 signaling, miR-342-5p, RAPGEF6, non-coding, MAPK.

Pasquinelli AE. MicroRNAs and their targets: recognition, regulation and an emerging reciprocal relationship. Nat Rev Genet 2012; 13(4): 271-82.
Friedman JM, Jones PA. MicroRNAs: critical mediators of differentiation, development and disease. Swiss Med Wkly 2009; 139(33-34): 466-72.
Garzon R, Calin GA, Croce CM. MicroRNAs in cancer. Annu Rev Med 2009; 60: 167-79.
Iuliano R, Vismara MF, Dattilo V, Trapasso F, Baudi F, Perrotti N. The role of microRNAs in cancer susceptibility. BioMed Res Int 2013; 2013: 591931.
Selbach M, Schwanhausser B, Thierfelder N, Fang Z, Khanin R, Rajewsky N. Widespread changes in protein synthesis induced by microRNAs. Nature 2008; 455(7209): 58-63.
Garzon R, Marcucci G, Croce CM. Targeting microRNAs in cancer: rationale, strategies and challenges. Nat Rev Drug Discov 2010; 9(10): 775-89.
Slamon DJ, Clark GM, Wong SG, Levin WJ, Ullrich A, McGuire WL. Human breast cancer: correlation of relapse and survival with amplification of the HER-2/neu oncogene. Science 1987; 235(4785): 177-82.
Gonzalez-Angulo AM, Litton JK, Broglio KR, et al. High risk of recurrence for patients with breast cancer who have human epidermal growth factor receptor 2-positive, node-negative tumors 1 cm or smaller. J Clin Oncol 2009; 27(34): 5700-6.
Leivonen SK, Sahlberg KK, Makela R, et al. High-throughput screens identify microRNAs essential for HER2 positive breast cancer cell growth. Mol Oncol 2014; 8(1): 93-104.
Kurebayashi J, Otsuki T, Tang CK, et al. Isolation and characterization of a new human breast cancer cell line, KPL-4, expressing the Erb B family receptors and interleukin-6. Br J Cancer 1999; 79(5-6): 707-17.
Lewis BP, Burge CB, Bartel DP. Conserved seed pairing, often flanked by adenosines, indicates that thousands of human genes are microRNA targets. Cell 2005; 120(1): 15-20.
Grimson A, Farh KK, Johnston WK, Garrett-Engele P, Lim LP, Bartel DP. MicroRNA targeting specificity in mammals: determinants beyond seed pairing. Mol Cell 2007; 27(1): 91-105.
Friedman RC, Farh KK, Burge CB, Bartel DP. Most mammalian mRNAs are conserved targets of microRNAs. Genome Res 2009; 19(1): 92-105.
Baek D, Villen J, Shin C, Camargo FD, Gygi SP, Bartel DP. The impact of microRNAs on protein output. Nature 2008; 455(7209): 64-71.
Maragkakis M, Reczko M, Simossis VA, et al. DIANA-microT web server: elucidating microRNA functions through target prediction. Nucleic Acids Res 2009; 37(Web server issue): W273-6.
Paraskevopoulou MD, Georgakilas G, Kostoulas N, et al. DIANAmicroT web server v5.0: service integration into miRNA functional analysis workflows. Nucleic Acids Res 2013; 41(Web server issue): W169-73.
Aure MR, Vitelli V, Jernstrom S, et al. Integrative clustering reveals a novel split in the luminal A subtype of breast cancer with impact on outcome. Breast Cancer Res 2017; 19(1): 44.
Curtis C, Shah SP, Chin SF, et al. The genomic and transcriptomic architecture of 2,000 breast tumours reveals novel subgroups. Nature 2012; 486(7403): 346-52.
Enerly E, Steinfeld I, Kleivi K, et al. miRNA-mRNA integrated analysis reveals roles for miRNAs in primary breast tumors. PLoS One 2011; 6(2): e16915.
Siendones E, SantaCruz-Calvo S, Martin-Montalvo A, et al. Membrane-bound CYB5R3 is a common effector of nutritional and oxidative stress response through FOXO3a and Nrf2. Antioxid Redox Signal 2014; 21(12): 1708-25.
Ran Q, Liang H, Gu M, et al. Transgenic mice overexpressing glutathione peroxidase 4 are protected against oxidative stress-induced apoptosis. J Biol Chem 2004; 279(53): 55137-46.
Cabreiro F, Picot CR, Perichon M, Castel J, Friguet B, Petropoulos I. Overexpression of mitochondrial methionine sulfoxide reductase B2 protects leukemia cells from oxidative stress-induced cell death and protein damage. J Biol Chem 2008; 283(24): 16673-81.
Sinha D, Srivastava S, Krishna L, D’Silva P. Unraveling the intricate organization of mammalian mitochondrial presequence translocases: existence of multiple translocases for maintenance of mitochondrial function. Mol Cell Biol 2014; 34(10): 1757-75.
Slamon DJ, Godolphin W, Jones LA, et al. Studies of the HER-2/neu proto-oncogene in human breast and ovarian cancer. Science 1989; 244(4905): 707-12.
Dawood S, Broglio K, Buzdar AU, Hortobagyi GN, Giordano SH. Prognosis of women with metastatic breast cancer by HER2 status and trastuzumab treatment: an institutional-based review. J Clin Oncol 2010; 28(1): 92-8.
Swain SM, Baselga J, Kim SB, et al. Pertuzumab, trastuzumab, and docetaxel in HER2-positive metastatic breast cancer. N Engl J Med 2015; 372(8): 724-34.
Yarden Y, Sliwkowski MX. Untangling the ErbB signalling network. Nat Rev Mol Cell Biol 2001; 2(2): 127-37.
Han JS, Crowe DL. Jun amino-terminal kinase 1 activation promotes cell survival in ErbB2-positive breast cancer. Anticancer Res 2010; 30(9): 3407-12.
Manole S, Richards EJ, Meyer AS. JNK pathway activation modulates acquired resistance to EGFR/HER2-targeted therapies. Cancer Res 2016; 76(18): 5219-28.
Phelps-Polirer K, Abt MA, Smith D, Yeh ES. Co-targeting of JNK and HUNK in resistant HER2-positive breast cancer. PLoS One 2016; 11(4): e0153025.
Phelps-Polirer K, Abt MA, Smith D, Yeh ES. Co-Targeting of JNK and HUNK in Resistant HER2-positive breast cancer. PLoS One 2016; 11(4): e0153025.
Gschwantler-Kaulich D, Grunt TW, Muhr D, Wagner R, Kolbl H, Singer CF. HER specific TKIs exert their antineoplastic effects on breast cancer cell lines through the involvement of STAT5 and JNK. PLoS One 2016; 11(1): e0146311.
Kyriakis JM, Banerjee P, Nikolakaki E, et al. The stress-activated protein-kinase subfamily of C-Jun kinases. Nature 1994; 369(6476): 156-60.
Kharbanda S, Saxena S, Yoshida K, et al. Translocation of SAPK/JNK to mitochondria and interaction with Bcl-x(L) in response to DNA damage. J Biol Chem 2000; 275(25): 19433.
Grazette LP, Matsui T, Rosenzweig A. Inhibition of ErbB2 causes mitochondrial dysfunction and impaired growth response in cardiomyocytes through altered Bcl-xS/Bcl-xL signaling: implications for herceptin-induced cardiomyopathy. Circulation 2004; 110(17): 8.
Gordon LI, Burke MA, Singh ATK, et al. Blockade of the ErbB2 receptor induces cardiomyocyte death through mitochondrial and reactive oxygen species-dependent pathways. J Biol Chem 2009; 284(4): 2080-7.
Liang H, Van Remmen H, Frohlich V, Lechleiter J, Richardson A, Ran Q. Gpx4 protects mitochondrial ATP generation against oxidative damage. Biochem Biophys Res Commun 2007; 356(4): 893-8.
Brigelius-Flohe R. Tissue-specific functions of individual gluta-thione peroxidases. Free Radic Biol Med 1999; 27(9-10): 951-65.
de Cabo R, Siendones E, Minor R, Navas P. CYB5R3: a key player in aerobic metabolism and aging? Aging (Albany NY) 2010; 2(1): 63-8.
Liu J, Lin AN. Role of JNK activation in apoptosis: a double-edged sword. Cell Res 2005; 15(1): 36-42.
Dube N, Kooistra MRH, Pannekoek WJ, et al. The RapGEF PDZ-GEF2 is required for maturation of cell-cell junctions. Cell Signal 2008; 20(9): 1608-15.
Severson EA, Lee WY, Capaldo CT, Nusrat A, Parkos CA. Junctional adhesion molecule A interacts with Afadin and PDZ-GEF2 to activate Rap1A, regulate beta1 integrin levels, and enhance cell migration. Mol Biol Cell 2009; 20(7): 1916-25.
Dvinge H, Git A, Graf S, et al. The shaping and functional consequences of the microRNA landscape in breast cancer. Nature 2013; 497(7449): 378-82.

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Article Details

Year: 2019
Published on: 06 December, 2018
Page: [155 - 165]
Pages: 11
DOI: 10.2174/2211536608666181206124922

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