Open Access

Epigenetically altered miR‑193a‑3p promotes HER2 positive breast cancer aggressiveness by targeting GRB7

  • Authors:
    • Yiyin Tang
    • Siyuan Yang
    • Maohua Wang
    • Dequan Liu
    • Yang Liu
    • Ying Zhang
    • Qian Zhang
  • View Affiliations

  • Published online on: April 15, 2019     https://doi.org/10.3892/ijmm.2019.4167
  • Pages: 2352-2360
  • Copyright: © Tang 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: )
Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )


Abstract

Emerging evidence has demonstrated that microRNAs (miRNAs/miRs) have various biological functions in the development of human epidermal growth factor receptor 2 (HER2) positive breast cancer. The aim of the present study is to reveal the mechanism of miR‑193a‑3p inhibiting the progress of HER2 positive breast cancer. The expression of miR‑193a‑3p was evaluated by quantitative polymerase chain reaction (PCR). The methylation status of miR‑193a‑3p was evaluated by PCR and pyrosequencing analysis. Overexpression of miR‑193a‑3p and growth factor receptor bound protein 7 (GRB7) combined with in vitro tumorigenic assays were conducted to determine the carcinostatic capacities of miR‑193a‑3p in HER2 positive breast cancer cells. The association between miR‑193a‑3p and GRB7 was determined by luciferase reporter assay. Protein level was evaluated using western blot analysis. miR‑193a‑3p was downregulated in HER2 positive breast cancer cells and clinical tissues. Methylation‑mediated silencing led to decreased expression of miR‑193a‑3p in HER2 positive breast cancer. Overexpression of miR‑193a‑3p could inhibit proliferation, migration and invasion of breast cancer cells. Overexpression of GRB7 could abolish this effect. miR‑193a‑3p could directly target the 3' untranslated region of GRB7. miR‑193a‑3p could directly or indirectly target extracellular signal‑regulated kinase 1/2 (ERK1/2) and forkhead box M1 (FOXM1) signaling. In conclusion, it was identified that silencing of miR‑193a‑3p through hypermethylation can promote HER2 positive breast cancer progress by targeting GRB7, ERK1/2 and FOXM1 signaling. The function of miR‑193a‑3p in HER2 positive breast cancer implicates its potential application in therapy.

References

1 

Siegel RL, Miller KD and Jemal A: Cancer statistics, 2018. CA Cancer J Clin. 68:7–30. 2018. View Article : Google Scholar : PubMed/NCBI

2 

Spizzo G, Obrist P, Ensinger C, Theurl I, Dünser M, Ramoni A, Gunsilius E, Eibl G, Mikuz G and Gastl G: Prognostic significance of Ep-CAM AND Her-2/neu overexpression in invasive breast cancer. Int J Cancer. 98:883–888. 2002. View Article : Google Scholar : PubMed/NCBI

3 

Walker RA: Immunohistochemical markers as predictive tools for breast cancer. J Clin Pathol. 61:689–696. 2008. View Article : Google Scholar

4 

Dunnwald LK, Rossing MA and Li CI: Hormone receptor status, tumor characteristics, and prognosis: A prospective cohort of breast cancer patients. Breast Cancer Res. 9:R62007. View Article : Google Scholar : PubMed/NCBI

5 

Iqbal N and Iqbal N: Human epidermal growth factor receptor 2 (HER2) in cancers: Overexpression and therapeutic implications. Mol Biol Int. 2014:8527482014. View Article : Google Scholar : PubMed/NCBI

6 

Roy AJ, Yankee RA, Brivkalns A and Fitch M: Viability of granulocytes obtained by filtration leukapheresis. Transfusion. 15:539–547. 1975. View Article : Google Scholar : PubMed/NCBI

7 

Singh R, Gupta S, Pawar SB, Pawar RS, Gandham SV and Prabhudesai S: Evaluation of ER, PR and HER-2 receptor expression in breast cancer patients presenting to a semi urban cancer centre in Western India. J Cancer Res Ther. 10:26–28. 2014. View Article : Google Scholar : PubMed/NCBI

8 

Kim JY, Jung WH and Koo JS: Expression of autophagy-related proteins according to androgen receptor and HER-2 status in estrogen receptor-negative breast cancer. PLoS One. 9:e1056662014. View Article : Google Scholar : PubMed/NCBI

9 

Slamon DJ, Godolphin W, Jones LA, Holt JA, Wong SG, Keith DE, Levin WJ, Stuart SG, Udove J and Ullrich A: Studies of the HER-2/neu proto-oncogene in human breast and ovarian cancer. Science. 244:707–712. 1989. View Article : Google Scholar : PubMed/NCBI

10 

Thompson SK, Sullivan TR, Davies R and Ruszkiewicz AR: Her-2/neu gene amplification in esophageal adenocarcinoma and its influence on survival. Ann Surg Oncol. 18:2010–2017. 2011. View Article : Google Scholar : PubMed/NCBI

11 

Sahlberg KK, Hongisto V, Edgren H, Mäkelä R, Hellström K, Due EU, Moen Vollan HK, Sahlberg N, Wolf M, Børresen- Dale AL, et al: The HER2 amplicon includes several genes required for the growth and survival of HER2 positive breast cancer cells. Mol Oncol. 7:392–401. 2013. View Article : Google Scholar

12 

Kauraniemi P and Kallioniemi A: Activation of multiple cancer-associated genes at the ERBB2 amplicon in breast cancer. Endocr Relat Cancer. 13:39–49. 2006. View Article : Google Scholar : PubMed/NCBI

13 

Valastyan S: Roles of microRNAs and other non-coding RNAs in breast cancer metastasis. J Mammary Gland Biol Neoplasia. 17:23–32. 2012. View Article : Google Scholar : PubMed/NCBI

14 

Zhang N, Wang X, Huo Q, Sun M, Cai C, Liu Z, Hu G and Yang Q: MicroRNA-30a suppresses breast tumor growth and metastasis by targeting metadherin. Oncogene. 33:3119–3128. 2014. View Article : Google Scholar

15 

Gregory PA, Bert AG, Paterson EL, Barry SC, Tsykin A, Farshid G, Vadas MA, Khew-Goodall Y and Goodall GJ: The miR-200 family and miR-205 regulate epithelial to mesenchymal transition by targeting ZEB1 and SIP1. Nat Cell Biol. 10:593–601. 2008. View Article : Google Scholar : PubMed/NCBI

16 

Pekow J, Meckel K, Dougherty U, Huang Y, Chen X, Almoghrabi A, Mustafi R, Ayaloglu-Butun F, Deng Z, Haider HI, et al: miR-193a-3p is a key tumor suppressor in ulcerative colitis-associated colon cancer and promotes carcinogenesis through upregulation of IL17RD. Clin Cancer Res. 23:5281–5291. 2017. View Article : Google Scholar : PubMed/NCBI

17 

Chou NH, Lo YH, Wang KC, Kang CH, Tsai CY and Tsai KW: MiR-193a-5p and -3p play a distinct role in gastric cancer: miR-193a-3p suppresses gastric cancer cell growth by targeting ETS1 and CCND1. Anticancer Res. 38:3309–3318. 2018. View Article : Google Scholar : PubMed/NCBI

18 

Bivin WW, Yergiyev O, Bunker ML, Silverman JF and Krishnamurti U: GRB7 expression and correlation with HER2 amplification in invasive breast carcinoma. Appl Immunohistochem Mol Morphol. 25:553–558. 2017. View Article : Google Scholar

19 

Nadler Y, González AM, Camp RL, Rimm DL, Kluger HM and Kluger Y: Growth factor receptor-bound protein-7 (Grb7) as a prognostic marker and therapeutic target in breast cancer. Ann Oncol. 21:466–473. 2010. View Article : Google Scholar :

20 

Lesurf R, Griffith OL, Griffith M, Hundal J, Trani L, Watson MA, Aft R, Ellis MJ, Ota D, Suman VJ, et al: Genomic characterization of HER2-positive breast cancer and response to neoadjuvant trastuzumab and chemotherapy-results from the ACOSOG Z1041 (Alliance) trial. Ann Oncol. 28:1070–1077. 2017. View Article : Google Scholar : PubMed/NCBI

21 

Lujambio A, Ropero S, Ballestar E, Fraga MF, Cerrato C, Setién F, Casado S, Suarez-Gauthier A, Sanchez-Cespedes M, Git A, et al: Genetic unmasking of an epigenetically silenced microRNA in human cancer cells. Cancer Res. 67:1424–1429. 2007. View Article : Google Scholar : PubMed/NCBI

22 

Saito Y, Liang G, Egger G, Friedman JM, Chuang JC, Coetzee GA and Jones PA: Specific activation of microRNA-127 with down-regulation of the proto-oncogene BCL6 by chromatin-modifying drugs in human cancer cells. Cancer Cell. 9:435–443. 2006. View Article : Google Scholar : PubMed/NCBI

23 

Yang H, Kong W, He L, Zhao JJ, O'Donnell JD, Wang J, Wenham RM, Coppola D, Kruk PA, Nicosia SV, et al: MicroRNA expression profiling in human ovarian cancer: miR-214 induces cell survival and cisplatin resistance by targetin PTEN. Cancer Res. 68:425–433. 2008. View Article : Google Scholar : PubMed/NCBI

24 

Fan Q, Hu X, Zhang H, Wang S, Zhang H, You C, Zhang CY, Liang H, Chen X and Ba Y: MiR-193a-3p is an important tumour suppressor in lung cancer and directly targets KRAS. Cell Physiol Biochem. 44:1311–1324. 2017. View Article : Google Scholar : PubMed/NCBI

25 

Wang Y, Chan DW, Liu VW, Chiu P and Ngan HY: Differential functions of growth factor receptor-bound protein 7 (GRB7) and its variant GRB7v in ovarian carcinogenesis. Clin Cancer Res. 16:2529–2539. 2010. View Article : Google Scholar : PubMed/NCBI

26 

Mak CS, Yung MM, Hui LM, Leung LL, Liang R, Chen K, Liu SS, Qin Y, Leung TH, Lee KF, et al: MicroRNA-141 enhances anoikis resistance in metastatic progression of ovarian cancer through targeting KLF12/Sp1/survivin axis. Mol Cancer. 16:112017. View Article : Google Scholar : PubMed/NCBI

27 

Stein D, Wu J, Fuqua SA, Roonprapunt C, Yajnik V, D'Eustachio P, Moskow JJ, Buchberg AM, Osborne CK and Margolis B: The SH2 domain protein GRB-7 is co-amplified, overexpressed and in a tight complex with HER2 in breast cancer. EMBO J. 13:1331–1340. 1994. View Article : Google Scholar : PubMed/NCBI

28 

Lamy PJ, Fina F, Bascoul-Mollevi C, Laberenne AC, Martin PM, Ouafik L and Jacot W: Quantification and clinical relevance of gene amplification at chromosome 17q12-q21 in human epidermal growth factor receptor 2-amplified breast cancers. Breast Cancer Res. 13:R152011. View Article : Google Scholar : PubMed/NCBI

29 

Bai T and Luoh SW: GRB-7 facilitates HER-2/Neu-mediated signal transduction and tumor formation. Carcinogenesis. 29:473–479. 2008. View Article : Google Scholar

30 

Meng F, Qian L, Lv L, Ding B, Zhou G, Cheng X, Niu S and Liang Y: miR-193a-3p regulation of chemoradiation resistance in oesophageal cancer cells via the PSEN1 gene. Gene. 579:139–145. 2016. View Article : Google Scholar : PubMed/NCBI

31 

Nie W, Ge HJ, Yang XQ, Sun X, Huang H, Tao X, Chen WS and Li B: LncRNA-UCA1 exerts oncogenic functions in non-small cell lung cancer by targeting miR-193a-3p. Cancer Lett. 371:99–106. 2016. View Article : Google Scholar

32 

Cui X, Chen X, Wang W, Chang A, Yang L, Liu C, Peng H, Wei Y, Liang W, Li S, et al: Epigenetic silencing of miR-203 in Kazakh patients with esophageal squamous cell carcinoma by MassARRAY spectrometry. Epigenetics. 12:698–707. 2017. View Article : Google Scholar : PubMed/NCBI

33 

Liang G and Weisenberger DJ: DNA methylation aberrancies as a guide for surveillance and treatment of human cancers. Epigenetics. 12:416–432. 2017. View Article : Google Scholar : PubMed/NCBI

34 

Chen K, Liu MX, Mak CS, Yung MM, Leung TH, Xu D, Ngu SF, Chan KK, Yang H, Ngan HY and Chan DW: Methylation-associated silencing of miR-193a-3p promotes ovarian cancer aggressiveness by targeting GRB7 and MAPK/ERK pathways. Theranostics. 8:423–436. 2018. View Article : Google Scholar :

35 

De Luca A, Maiello MR, D'Alessio A, Pergameno M and Normanno N: The RAS/RAF/MEK/ERK and the PI3K/AKT signalling pathways: Role in cancer pathogenesis and implications for therapeutic approaches. Expert Opin Ther Targets. 16(Suppl 2): S17–S27. 2012. View Article : Google Scholar : PubMed/NCBI

36 

Chu PY, Li TK, Ding ST, Lai IR and Shen TL: EGF-induced Grb7 recruits and promotes Ras activity essential for the tumorigenicity of Sk-Br3 breast cancer cells. J Biol Chem. 285:29279–29285. 2010. View Article : Google Scholar : PubMed/NCBI

37 

Laoukili J, Stahl M and Medema RH: FoxM1: At the crossroads of ageing and cancer. Biochim Biophys Acta. 1775:92–102. 2007.

38 

Lüscher-Firzlaff JM, Lilischkis R and Lüscher B: Regulation of the transcription factor FOXM1c by Cyclin E/CDK2. FEBS Lett. 580:1716–1722. 2006. View Article : Google Scholar : PubMed/NCBI

39 

Major ML, Lepe R and Costa RH: Forkhead box M1B transcriptional activity requires binding of Cdk-cyclin complexes for phosphorylation-dependent recruitment of p300/CBP coactiva-tors. Mol Cell Biol. 24:2649–2661. 2004. View Article : Google Scholar : PubMed/NCBI

40 

Francis RE, Myatt SS, Krol J, Hartman J, Peck B, McGovern UB, Wang J, Guest SK, Filipovic A, Gojis O, et al: FoxM1 is a downstream target and marker of HER2 overexpression in breast cancer. Int J Oncol. 35:57–68. 2009.PubMed/NCBI

Related Articles

Journal Cover

June 2019
Volume 43 Issue 6

Print ISSN: 1107-3756
Online ISSN:1791-244X

Sign up for eToc alerts

Recommend to Library

Copy and paste a formatted citation
APA
Tang, Y., Yang, S., Wang, M., Liu, D., Liu, Y., Zhang, Y., & Zhang, Q. (2019). Epigenetically altered miR‑193a‑3p promotes HER2 positive breast cancer aggressiveness by targeting GRB7. International Journal of Molecular Medicine, 43, 2352-2360. https://doi.org/10.3892/ijmm.2019.4167
MLA
Tang, Y., Yang, S., Wang, M., Liu, D., Liu, Y., Zhang, Y., Zhang, Q."Epigenetically altered miR‑193a‑3p promotes HER2 positive breast cancer aggressiveness by targeting GRB7". International Journal of Molecular Medicine 43.6 (2019): 2352-2360.
Chicago
Tang, Y., Yang, S., Wang, M., Liu, D., Liu, Y., Zhang, Y., Zhang, Q."Epigenetically altered miR‑193a‑3p promotes HER2 positive breast cancer aggressiveness by targeting GRB7". International Journal of Molecular Medicine 43, no. 6 (2019): 2352-2360. https://doi.org/10.3892/ijmm.2019.4167