Cantharidin modulates the E2F1/MCM7‑miR‑106b‑93/p21‑PTEN signaling axis in MCF‑7 breast cancer cells

Zhang,Hui; Yan,Xiuli

doi:10.3892/ol.2015.3681

Cantharidin modulates the E2F1/MCM7‑miR‑106b‑93/p21‑PTEN signaling axis in MCF‑7 breast cancer cells

Authors:
- Hui Zhang
- Xiuli Yan
View Affiliations

Affiliations: Research Center for Traditional Chinese Medicine Complexity System, Shanghai University of Traditional Chinese Medicine, Shanghai 201203, P.R. China, Internal Medicine of Traditional Chinese Medicine, Yueyang Hospital of Integrated Traditional Chinese and Western Medicine, Shanghai University of Traditional Chinese Medicine, Shanghai 200437, P.R. China
Published online on: September 7, 2015 https://doi.org/10.3892/ol.2015.3681
Pages: 2849-2855
Copyright: © Zhang 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

Cantharidin (CTD) is one of numerous natural products used in traditional Chinese medicine for the treatment of cancer. The aim of the present study was to investigate the effects of CTD on changes in the expression of microRNAs (miRNAs/miRs) and to explore its anti‑proliferative effect on MCF‑7 breast cancer cells. The proliferation of MCF‑7 cells was measured by performing an MTT assay. MCF‑7 cells were treated with various concentrations of CTD for 48 h, and the expression profiles of miRNAs in CTD‑treated and ‑untreated MCF‑7 cells were detected using miRNA microarray chips. The array data were confirmed by reverse transcription‑quantitative polymerase chain reaction and protein expression levels were measured by western blot analysis. The 50% inhibitory concentration of CTD was 1.75 µg/ml following treatment for 48 h and CTD significantly inhibited the proliferation of MCF‑7 cells in a dose‑dependent manner (P<0.01). Furthermore, microarray analysis identified 35 miRNAs that were up‑regulated (fold change ≥2.0 and P<0.01) and 45 miRNAs that were down‑regulated (fold change ≤ 0.5 and P<0.01) in response to CTD treatment. Thus, numerous CTD‑induced miRNAs appeared to be associated with breast cancer. Notably, CTD repressed the expression of miR‑106b‑93, its host gene MCM7 and its transcription factor E2F1. In addition, CTD induced an increase in the protein expression levels of miR‑106b‑93 target genes p21 and phosphatase and tensin homolog. These observations suggested that the modulation of miRNA expression is an important mechanism underlying the biological effects of CTD in breast cancer.

View Figures

View References

1	Filipowicz W, Bhattacharyya SN and Sonenberg N: Mechanisms of post-transcriptional regulation by microRNAs: Are the answers in sight? Nat Rev Genet. 9:102–114. 2008. View Article : Google Scholar : PubMed/NCBI
2	Griffiths-Jones S, Saini HK, van Dongen S and Enright AJ: miRBase: Tools for microRNA genomics. Nucleic Acids Res. 36:D154–D158. 2008. View Article : Google Scholar : PubMed/NCBI
3	Johnson SM, Grosshans H, Shingara J, Byrom M, Jarvis R, Cheng A, Labourier E, Reinert KL, Brown D and Slack FJ: RAS is regulated by the let-7 microRNA family. Cell. 120:635–647. 2005. View Article : Google Scholar : PubMed/NCBI
4	Bonci D, Coppola V, Musumeci M, Addario A, Giuffrida R, Memeo L, D'Urso L, Pagliuca A, Biffoni M, Labbaye C, et al: The miR-15a-miR-16-1 cluster controls prostate cancer by targeting multiple oncogenic activities. Nat Med. 14:1271–1277. 2008. View Article : Google Scholar : PubMed/NCBI
5	Si ML, Zhu S, Wu H, Lu Z, Wu F and Mo YY: MiR-21-mediated tumor growth. Oncogene. 26:2799–2803. 2007. View Article : Google Scholar : PubMed/NCBI
6	Lee DY, Deng Z, Wang CH and Yang BB: MicroRNA-378 promotes cell survival, tumor growth and angiogenesis by targeting SuFu and Fus-1 expression. Proc Natl Acad Sci (USA). 104:20350–20355. 2007. View Article : Google Scholar : PubMed/NCBI
7	Esquela-Kerscher A and Slack FJ: Oncomirs: microRNAs with a role in cancer. Nat Rev Cancer. 6:259–269. 2006. View Article : Google Scholar : PubMed/NCBI
8	Hossain A, Kuo MT and Saunders GF: Mir-17-5p regulates breast cancer cell proliferation by inhibiting translation of AIB1 mRNA. Mol Cell Biol. 26:8191–8201. 2006. View Article : Google Scholar : PubMed/NCBI
9	Ma L, Teruya-Feldstein J and Weinberg RA: Tumour invasion and metastasis initiated by microRNA-10b in breast cancer. Nature. 449:682–688. 2007. View Article : Google Scholar : PubMed/NCBI
10	Frankel LB, Christoffersen NR, Jacobsen A, Lindow M, Krogh A and Lund AH: Programmed cell death 4 (PDCD4) is an important functional target of the microRNA miR-21 in breast cancer cells. J Biol Chem. 283:1026–1033. 2007. View Article : Google Scholar : PubMed/NCBI
11	Wang V and Wu W: MicroRNA: A new player in breast cancer development (Review). Cancer Mol. 3:133–138. 2007.
12	Liu G, Wong-Staal F and Li QX: Development of new RNAi therapeutics. Histol Histopathol. 22:211–217. 2007.PubMed/NCBI
13	Miller BA, Chu KC, Hankey BF and Ries LA: Cancer incidence and mortality patterns among specific Asian and pacific islander populations in the U.S. Cancer Causes Control. 19:227–256. 2008. View Article : Google Scholar : PubMed/NCBI
14	Efferth T, Rauh R, Kahl S, Tomicic M, Böchzelt H, Tome ME, Briehl MM, Bauer R and Kaina B: Molecular modes of action of cantharidin in tumor cells. Biochem Pharmacol. 69:811–818. 2005. View Article : Google Scholar : PubMed/NCBI
15	Zheng LH, Bao YL, Wu Y, Yu CL, Meng X and Li YX: Cantharidin reverses multidrug resistance of human hepatoma HepG2/ADM cells via down-regulation of P-glycoprotein expression. Cancer Lett. 272:102–109. 2008. View Article : Google Scholar : PubMed/NCBI
16	Sagawa M, Nakazato T, Uchida H, Ikeda Y and Kizaki M: Cantharidin induces apoptosis of human multiple myeloma cells via inhibition of the JAK/STAT pathway. Cancer Sci. 99:1820–1826. 2008. View Article : Google Scholar : PubMed/NCBI
17	Shivdasani RA: MicroRNAs: Regulators of gene expression and cell differentiation. Blood. 108:3646–3653. 2006. View Article : Google Scholar : PubMed/NCBI
18	Ross JS, Carlson JA and Brock G: miRNA: The new gene silencer. Am J Clin Pathol. 128:830–836. 2007. View Article : Google Scholar : PubMed/NCBI
19	Chen CZ: MicroRNAs as oncogenes and tumor suppressors. N Engl J Med. 353:1768–1771. 2005. View Article : Google Scholar : PubMed/NCBI
20	Reddy SD, Ohshiro K, Rayala SK and Kumar R: MicroRNA-7, a homeobox D10 target, inhibits p21-activated kinase 1 and regulates its functions. Cancer Res. 68:8195–8200. 2008. View Article : Google Scholar : PubMed/NCBI
21	Li D, Zhao Y, Liu C, Chen X, Qi Y, Jiang Y, Zou C, Zhang X, Liu S, Wang X, et al: Analysis of MiR-195 and MiR-497 expression, regulation and role in breast cancer. Clin Cancer Res. 17:1722–1730. 2011. View Article : Google Scholar : PubMed/NCBI
22	Wang C, Zheng X, Shen C and Shi Y: MicroRNA-203 suppresses cell proliferation and migration by targeting BIRC5 and LASP1 in human triple-negative breast cancer cells. J Exp Clin Cancer Res. 31:582012. View Article : Google Scholar : PubMed/NCBI
23	Derfoul A, Juan AH, Difilippantonio MJ, Palanisamy N, Ried T and Sartorelli V: Decreased microRNA-214 levels in breast cancer cells coincides with increased cell proliferation, invasion and accumulation of the Polycomb Ezh2 methyltransferase. Carcinogenesis. 32:1607–1614. 2011. View Article : Google Scholar : PubMed/NCBI
24	Ivanovska I, Ball AS, Diaz RL, Magnus JF, Kibukawa M, Schelter JM, Kobayashi SV, Lim L, Burchard J, Jackson AL, et al: MicroRNAs in the miR-106b family regulate p21/CDKN1A and promote cell cycle progression. Mol Cell Biol. 28:2167–2174. 2008. View Article : Google Scholar : PubMed/NCBI
25	Feng M, Li Z, Aau M, Wong CH, Yang X and Yu Q: Myc/miR-378/TOB2/cyclin D1 functional module regulates oncogenic transformation. Oncogene. 30:2242–2251. 2011. View Article : Google Scholar : PubMed/NCBI
26	Li Y, Tan W, Neo TW, Aung MO, Wasser S, Lim SG and Tan TM: Role of the miR-106b-25 microRNA cluster in hepatocellular carcinoma. Cancer Sci. 100:1234–1242. 2009. View Article : Google Scholar : PubMed/NCBI
27	Kan T, Sato F, Ito T, Matsumura N, David S, Cheng Y, Agarwal R, Paun BC, Jin Z, Olaru AV, et al: The miR-106b-25 polycistron, activated by genomic amplification, functions as an oncogene by suppressing p21 and Bim. Gastroenterology. 136:1689–1700. 2009. View Article : Google Scholar : PubMed/NCBI
28	Hui AB, Lenarduzzi M, Krushel T, Waldron L, Pintilie M, Shi W, Perez-Ordonez B, Jurisica I, O'Sullivan B, Waldron J, et al: Comprehensive MicroRNA profiling for head and neck squamous cell carcinomas. Clin Cancer Res. 16:1129–1139. 2010. View Article : Google Scholar : PubMed/NCBI
29	Poliseno L, Salmena L, Riccardi L, Fornari A, Song MS, Hobbs RM, Sportoletti P, Varmeh S, Egia A, Fedele G, et al: Identification of the miR-106b~25 microRNA cluster as a proto-oncogenic PTEN-targeting intron that cooperates with its host gene MCM7 in transformation. Sci Signal. 3:ra292010.PubMed/NCBI
30	Petrocca F, Visone R, Onelli MR, Shah MH, Nicoloso MS, de Martino I, Iliopoulos D, Pilozzi E, Liu CG, Negrini M, et al: E2F1-Regulated MicroRNAs impair TGFbeta-dependent cell-cycle arrest and apoptosis in gastric cancer. Cancer Cell. 13:272–286. 2008. View Article : Google Scholar : PubMed/NCBI
31	Han S, Park K, Bae BN, Kim KH, Kim HJ, Kim YD and Kim HY: E2F1 expression is related with the poor survival of lymph node-positive breast cancer patients treated with fluorouracil, doxorubicin and cyclophosphamide. Breast Cancer Res Treat. 82:11–16. 2003. View Article : Google Scholar : PubMed/NCBI