miR‑135b‑5p enhances the sensitivity of HER‑2 positive breast cancer to trastuzumab via binding to cyclin D2

Li,Zhilan; Qin,Yiyu; Chen,Peihong; Luo,Qiong; Shi,Haiyan; Jiang,Xiudi

doi:10.3892/ijmm.2020.4681

miR‑135b‑5p enhances the sensitivity of HER‑2 positive breast cancer to trastuzumab via binding to cyclin D2

Authors:
- Zhilan Li
- Yiyu Qin
- Peihong Chen
- Qiong Luo
- Haiyan Shi
- Xiudi Jiang
View Affiliations

Affiliations: Department of Clinical Laboratory, Seventh People's Hospital of Shanghai University of Traditional Chinese Medicine, Shanghai 200137, P.R. China, School of Clinical Medicine, Jiangsu Medical Vocational College, Yancheng, Jiangsu 224005, P.R. China
Published online on: July 22, 2020 https://doi.org/10.3892/ijmm.2020.4681
Pages: 1514-1524
Copyright: © Li 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

Trastuzumab has led to a marked improvement in the outcomes of patients with human epidermal growth factor receptor 2 (HER‑2)‑positive breast cancer. However, the effects of trastuzumab on HER‑2‑positive breast cancer are limited by the emergence of its cardiotoxicside effects. MicroRNA (miR)‑135b‑5p has been shown to inhibit tumor metastasis in breast cancer. The present study aimed to explore the effects of miR‑135b‑5p overexpression on the efficacy of trastuzumab in HER‑2‑positive breast cancer. Reverse transcription‑quantitative PCR was performed to detect the levels of miR‑135b‑5p. Cell viability was evaluated with a Cell Counting Kit‑8 assay. Annexin V/propidium iodide staining was employed to detect the number of apoptotic cells. Flow cytometry assay was performed to investigate the cell cycle. Western blotting was used to detect the expression levels of Bax, cleaved caspase‑3, Bcl‑2, cyclin D2, p27Kip1 and cyclin E1. Cell migration and invasion were detected by Transwell assay. Luciferase assays were conducted to identify the target gene of miR‑135b‑5p. In addition, an in vivo tumor xenograft model was established. miR‑135b‑5p agomir significantly enhanced the anti‑proliferative effect of trastuzumab on HER‑2‑positive breast cancer cells via the induction of apoptosis, whereas the anti‑metastatic effect of trastuzumab was enhanced by miR‑135b‑5p agomir treatment. Subsequently, luciferase assays indicated that cyclin D2 was the direct target of miR‑135b‑5p, whereas overexpression of the latter arrested cell cycleduring the G0/G1 phase. Moreover, miR‑135b‑5p agomir notably increased the antitumor effect of trastuzumab in vivo. The data demonstrated that miR‑135b‑5p sensitized HER‑2‑positive breast cancer cells to trastuzumab in vitro and in vivo by directly binding to cyclin D2. These results suggested that the combination of miR‑135b‑5p with trastuzumab may be a therapeutic strategy for patients with HER‑2‑positive breast cancer.

View Figures

View References

1	Tormo E, Adam-Artigues A, Ballester S, Pineda B, Zazo S, González-Alonso P, Albanell J, Rovira A, Rojo F, Lluch A and Eroles P: The role of miR-26a and miR-30b in HER2⁺ breast cancer trastuzumab resistance and regulation of the CCNE2 gene. Sci Rep. 7:413092017. View Article : Google Scholar
2	Akram M, Iqbal M, Daniyal M and Khan AU: Awareness and current knowledge of breast cancer. Biol Res. 50:332017. View Article : Google Scholar : PubMed/NCBI
3	Merino Bonilla JA, Torres Tabanera M and Ros Mendoza LH: Breast cancer in the 21st century: From early detection to new therapies. Radiologia. 59:368–379. 2017.In English, Spanish. View Article : Google Scholar : PubMed/NCBI
4	Kim YJ, Jung SY and Kim K: Survival benefit of radiotherapy after surgery in de novo stage IV breast cancer: A population-based propensity-score matched analysis. Sci Rep. 9:85272019. View Article : Google Scholar : PubMed/NCBI
5	Nguyen PL, Taghian AG, Katz MS, Niemierko A, Raad RF, Boon WL, Bellon JR, Wong JS, Smith BL and Harris JR: Breast cancer subtype approximated by estrogen receptor, progesterone receptor, and HER-2 is associated with local and distant recurrence after breast-conserving therapy. J Clin Oncol. 26:2373–2378. 2008. View Article : Google Scholar : PubMed/NCBI
6	Chavez-Blanco A, Perez-Sanchez V, Gonzalez-Fierro A, Vela-Chavez T, Candelaria M, Cetina L, Vidal S and Dueñas-Gonzalez A: HER2 expression in cervical cancer as a potential therapeutic target. BMC Cancer. 4:592004. View Article : Google Scholar : PubMed/NCBI
7	Baselga J and Swain SM: Novel anticancer targets: Revisiting ERBB2 and discovering ERBB3. Nat Rev Cancer. 9:463–475. 2009. View Article : Google Scholar : PubMed/NCBI
8	Horton J: Trastuzumab use in breast cancer: Clinical issues. Cancer Control. 9:499–507. 2002. View Article : Google Scholar
9	Sarup JC, Johnson RM, King KL, Fendly BM, Lipari MT, Napier MA, Ullrich A and Shepard HM: Characterization of an anti-p185HER2 monoclonal antibody that stimulates receptor function and inhibits tumor cell growth. Growth Regul. 1:72–82. 1991.PubMed/NCBI
10	Lane HA, Beuvink I, Motoyama AB, Daly JM, Neve RM and Hynes NE: ErbB2 potentiates breast tumor proliferation through modulation of p27(Kip1)-Cdk2 complex formation: Receptor overexpression does not determine growth dependency. Mol Cell Biol. 20:3210–3223. 2000. View Article : Google Scholar : PubMed/NCBI
11	Petit AM, Rak J, Hung MC, Rockwell P, Goldstein N, Fendly B and Kerbel RS: Neutralizing antibodies against epidermal growth factor and ErbB-2/neu receptor tyrosine kinases down-regulate vascular endothelial growth factor production by tumor cells in vitro and in vivo: Angiogenic implications for signal transduction therapy of solid tumors. Am J Pathol. 151:1523–1530. 1997.PubMed/NCBI
12	Sliwkowski MX, Lofgren JA, Lewis GD, Hotaling TE, Fendly BM and Fox JA: Nonclinical studies addressing the mechanism of action of trastuzumab (Herceptin). Semin Oncol. 26:60–70. 1999.PubMed/NCBI
13	Van Swearingen AED, Siegel MB, Deal AM, Sambade MJ, Hoyle A, Hayes DN, Jo H, Little P, Dees EC, Muss H, et al: LCCC 1025: A phase II study of everolimus, trastuzumab, and vinorelbine to treat progressive HER2-positive breast cancer brain metastases. Breast Cancer Res Treat. 171:637–648. 2018. View Article : Google Scholar : PubMed/NCBI
14	Ding K, Wu Z, Li X, Sheng Y, Wang X and Tan S: LMO4 mediates trastuzumab resistance in HER2 positive breast cancer cells. Am J Cancer Res. 8:594–609. 2018.PubMed/NCBI
15	Tan H, Huang S, Zhang Z, Qian X, Sun P and Zhou X: Pan-Cancer analysis on microRNA-associated gene activation. EBioMedicine. 43:82–97. 2019. View Article : Google Scholar : PubMed/NCBI
16	Bartel DP: Metazoan MicroRNAs. Cell. 173:20–51. 2018. View Article : Google Scholar : PubMed/NCBI
17	Schneider A, Victoria B, Lopez YN, Suchorska W, Barczak W, Sobecka A, Golusinski W, Masternak MM and Golusinski P: Tissue and serum microRNA profile of oral squamous cell carcinoma patients. Sci Rep. 8:6752018. View Article : Google Scholar : PubMed/NCBI
18	Venturutti L, Cordo Russo RI, Rivas MA, Mercogliano MF, Izzo F, Oakley RH, Pereyra MD, Proietti CJ, Yankilevich P, Roa JC, et al: miR-16 mediates trastuzumab and lapatinib response in ErbB-2-positive breast and gastric cancer via its novel targets CCNJ and FUBP1. Oncogene. 35:6189–6202. 2016. View Article : Google Scholar : PubMed/NCBI
19	Song W, Wu S, Wu Q, Zhou L, Yu L, Zhu B and Gong X: The microRNA-141-3p/CDK8 pathway regulates the chemosensitivity of breast cancer cells to trastuzumab. J Cell Biochem. 120:14095–14106. 2019. View Article : Google Scholar : PubMed/NCBI
20	Noyan S, Gurdal H and Gur Dedeoglu B: Involvement of miR-770-5p in trastuzumab response in HER2 positive breast cancer cells. PLoS One. 14:e02158942019. View Article : Google Scholar : PubMed/NCBI
21	De Cola A, Volpe S, Budani MC, Ferracin M, Lattanzio R, Turdo A, D'Agostino D, Capone E, Stassi G, Todaro M, et al: miR-205-5p-mediated downregulation of ErbB/HER receptors in breast cancer stem cells results in targeted therapy resistance. Cell Death Dis. 6:e18232015. View Article : Google Scholar : PubMed/NCBI
22	Pu T, Shen M, Li S, Yang L, Gao H, Xiao L, Zhong X, Zheng H, Liu Y, Ye F and Bu H: Repression of miR-135b-5p promotes metastasis of early-stage breast cancer by regulating downstream target SDCBP. Lab Invest. 99:1296–1308. 2019. View Article : Google Scholar : PubMed/NCBI
23	Zhang Y, Xia F, Zhang F, Cui Y, Wang Q, Liu H and Wu Y: miR-135b-5p enhances doxorubicin-sensitivity of breast cancer cells through targeting anterior gradient 2. J Exp Clin Cancer Res. 38:262019. View Article : Google Scholar : PubMed/NCBI
24	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
25	National Institutes of Health: Guide for the Care and Use of Laboratory Animals. The National Academies Press; Washington, DC: pp. p2462011
26	Workman P, Aboagye EO, Balkwill F, Balmain A, Bruder G, Chaplin DJ, Double JA, Everitt J, Farningham DAH, Glennie MJ, et al: Guidelines for the welfare and use of animals in cancer research. Br J Cancer. 102:1555–1577. 2010. View Article : Google Scholar : PubMed/NCBI
27	Le XF, Almeida MI, Mao W, Spizzo R, Rossi S, Nicoloso MS, Zhang S, Wu Y, Calin GA and Bast RC Jr: Modulation of MicroRNA-194 and cell migration by HER2-targeting trastuzumab in breast cancer. PLoS One. 7:e411702012. View Article : Google Scholar : PubMed/NCBI
28	Lulli V, Buccarelli M, Martini M, Signore M, Biffoni M, Giannetti S, Morgante L, Marziali G, Ilari R, Pagliuca A, et al: miR-135b suppresses tumorigenesis in glioblastoma stem-like cells impairing proliferation, migration and self-renewal. Oncotarget. 6:37241–37256. 2015. View Article : Google Scholar : PubMed/NCBI
29	Xiao M, Cai J, Cai L, Jia J, Xie L, Zhu Y, Huang B, Jin D and Wang Z: Let-7e sensitizes epithelial ovarian cancer to cisplatin through repressing DNA double strand break repair. J Ovarian Res. 10:242017. View Article : Google Scholar : PubMed/NCBI
30	Krützfeldt J, Rajewsky N, Braich R, Rajeev KG, Tuschl T, Manoharan M and Stoffel M: Silencing of microRNAs in vivo with 'antagomirs'. Nature. 438:685–689. 2005. View Article : Google Scholar : PubMed/NCBI
31	Tang H, Song C, Ye F, Gao G, Ou X, Zhang L and Xie X and Xie X: miR-200c suppresses stemness and increases cellular sensitivity to trastuzumab in HER2⁺ breast cancer. J Cell Mol Med. 23:8114–8127. 2019. View Article : Google Scholar : PubMed/NCBI
32	Montagnoli A, Fiore F, Eytan E, Carrano AC, Draetta GF, Hershko A and Pagano M: Ubiquitination of p27 is regulated by Cdk-dependent phosphorylation and trimeric complex formation. Genes Dev. 13:1181–1189. 1999. View Article : Google Scholar : PubMed/NCBI
33	Zhou J, Tian Y, Li J, Lu B, Sun M, Zou Y, Kong R, Luo Y, Shi Y, Wang K and Ji G: miR-206 is down-regulated in breast cancer and inhibits cell proliferation through the up-regulation of cyclinD2. Biochem Biophys Res Commun. 433:207–212. 2013. View Article : Google Scholar : PubMed/NCBI
34	Vu T and Claret FX: Trastuzumab: Updated mechanisms of action and resistance in breast cancer. Front Oncol. 2:622012. View Article : Google Scholar : PubMed/NCBI
35	Yu WF, Wang HM, Lu BC, Zhang GZ, Ma HM and Wu ZY: miR-206 inhibits human laryngeal squamous cell carcinoma cell growth by regulation of cyclin D2. Eur Rev Med Pharmacol Sci. 19:2697–2702. 2015.
36	Wu X, Zeng Y, Wu S, Zhong J, Wang Y and Xu J: miR-204, down-regulated in retinoblastoma, regulates proliferation and invasion of human retinoblastoma cells by targeting cyclin D2 and MMP-9. FEBS Lett. 589:645–650. 2015. View Article : Google Scholar : PubMed/NCBI
37	Pan JL, Yuan DZ, Zhao YB, Nie L, Lei Y, Liu M, Long Y, Zhang JH, Blok LJ, Burger CW and Yue LM: Progesterone-induced miR-133a inhibits the proliferation of endometrial epithelial cells. Acta Physiol (Oxf). 219:683–692. 2017. View Article : Google Scholar
38	Zhao S, Han J, Zheng L, Yang Z, Zhao L and Lv Y: MicroRNA-203 regulates growth and metastasis of breast cancer. Cell Physiol Biochem. 37:35–42. 2015. View Article : Google Scholar : PubMed/NCBI