Deregulation of miR-21 and miR-155 and their putative targets after silibinin treatment in T47D breast cancer cells

Document Type : Original Article

Authors

1 Department of Cell and Molecular Biology, School of Biology, College of Science, University of Tehran, Tehran, Iran

2 Department of Genetics, College of Science, Rasht Branch, Islamic Azad University, Rasht, Iran

Abstract

Objective(s):MicroRNAs (miRNAs) are a class of short RNAs that control the biological processes including cell proliferation, apoptosis and development. Aberrant expression of miRNAs was determined in the different stages of tumor development and metastasis. To study the effect of silibinin on miRNAs expression, we evaluated quantitative expression of miR-21 and miR-155 as two oncomiRs and several potential targets in silibinin-treated T47D cells.
Materials and Methods:The rate of proliferation and apoptosis was measured in silibinin-treated and untreated cells. The expression levels of miR-21 and miR-155 were evaluated in T47D cells treated with silibinin (100 µg/ml). Also, their putative targets were predicted in apoptotic pathways using multiple algorithms; as a confirmation, the transcription level of APAF-1, CASP-9 and BID was evaluated.
Results:In silibinin-treated cells, death was occurred in a dose and time-dependent manner. miR-21 and miR-155 was downregulated in cells treated with silibinin (100 µg/ml). It is noticeable that the expression of their potential targets including CASP-9 and APAF-1 was increased in silibinin-treated cells after 48 hr.
Conclusion:Our findings showed a correlation between the expression of miR-21 and miR-155 and apoptosis in silibinin treated T47D cells. It seems that miRNAs such as miR-21 and miR-155 were regulated by silibinin. Also, increase in the transcript level of APAF-1 and CASP-9 after downregulation of miR-21 and miR-155 might indicate that these genes were targeted by aforementioned miRNAs in T47D cells.

Keywords

References

1. Tiwari P, Kumar A, Balakrishnan S, Kushwaha HS, Mishra KP. Silibinin-induced apoptosis in MCF7 and T47D human breast carcinoma cells involves caspase-8 activation and mitochondrial pathway. Cancer Invest 2011; 29:12-20.
2. Noh EM, Yi MS, Youn HJ, Lee BK, Lee YR, Han JH, et al. Silibinin enhances ultraviolet B-induced apoptosis in mcf-7 human breast cancer cells. J Breast Cancer 2011; 14:8-13.
3. Lee SO, Jeong YJ, Im HG, Kim CH, Chang YC, Lee IS. Silibinin suppresses PMA-induced MMP-9 expression by blocking the AP-1 activation via MAPK signaling pathways in MCF-7 human breast carcinoma cells. Biochem Biophys Res Commun 2007; 354:165-171.
4. Nasiri M, Zarghami N, Koshki KN, Mollazadeh M, Moghaddam MP, Yamchi MR, et al. Curcumin and silibinin inhibit telomerase expression in T47D human breast cancer cells. Asian Pac J Cancer Prev 2013; 14:3449-3453.
5. Hsieh YS, Chu SC, Yang SF, Chen PN, Liu YC, Lu KH. Silibinin suppresses human osteosarcoma MG-63 cell invasion by inhibiting the ERK-dependent c-Jun/AP-1 induction of MMP-2. Carcinogenesis 2007; 28:977-987.
6. Ranji N, Sadeghizadeh M, Shokrgozar MA, Bakhshandeh B, Karimipour M, Amanzadeh A, et al. MiR-17-92 cluster: an apoptosis inducer or proliferation enhancer. Mol Cell Biochem 2013; 380:229-238.
7. Calin GA, Croce CM. MicroRNA signatures in human cancers. Nat Rev Cancer 2006; 6:857-866.
8. Tili E, Michaille JJ, Calin GA. Expression and function of micro-RNAs in immune cells during normal or disease state. Int J Med Sci 2008; 5:73-79.
9. Selbach M, Schwanhausser B, Thierfelder N, Fang Z, Khanin R, Rajewsky N. Widespread changes in protein synthesis induced by microRNAs. Nature 2008; 455:58-63.
10. Ruan K, Fang X, Ouyang G. MicroRNAs: novel regulators in the hallmarks of human cancer. Cancer Lett 2009; 285:116-126.
11. Lechman ER, Gentner B, van Galen P, Giustacchini A, Saini M, Boccalatte FE, et al. Attenuation of miR-126 activity expands HSC in vivo without exhaustion. Cell Stem Cell 2012; 11:799-811.
12. Shimizu S, Takehara T, Hikita H, Kodama T, Miyagi T, Hosui A, et al. The let-7 family of microRNAs inhibits Bcl-xL expression and potentiates sorafenib-induced apoptosis in human hepatocellular carcinoma. J Hepatol 2010; 52:698-704.
13. Yang M, Li Y, Padgett RW. MicroRNAs: Small regulators with a big impact. Cytokine Growth Factor Rev 2005; 16:387-393.
14. Wang Z, Li Y, Kong D, Ahmad A, Banerjee S, Sarkar FH. Cross-talk between miRNA and Notch signaling pathways in tumor development and progression. Cancer Lett 2010; 292:141-148.
15. Lal A, Navarro F, Maher CA, Maliszewski LE, Yan N, O'Day E, et al. miR-24 Inhibits cell proliferation by targeting E2F2, MYC, and other cell-cycle genes via binding to "seedless" 3'UTR microRNA recognition elements. Mol Cell 2009; 35:610-625.
16. Cho WC. OncomiRs: the discovery and progress of microRNAs in cancers. Mol Cancer 2007; 6:60.
17. Johnson CD, Esquela-Kerscher A, Stefani G, Byrom M, Kelnar K, Ovcharenko D, et al. The let-7 microRNA represses cell proliferation pathways in human cells. Cancer Res 2007; 67:7713-7722.
18. Sarkar FH, Li Y, Wang Z, Kong D, Ali S. Implication of microRNAs in drug resistance for designing novel cancer therapy. Drug Resist Updat 2010; 13:57-66.
19. Yang H, Kong W, He L, Zhao JJ, O'Donnell JD, Wang J, et al. MicroRNA expression profiling in human ovarian cancer: miR-214 induces cell survival and cisplatin resistance by targeting PTEN. Cancer Res 2008; 68:425-433.
20. Chen YH, Chen CL, Liang CM, Liang JB, Tai MC, Chang YH, et al. Silibinin inhibits ICAM-1 expression via regulation of N-linked and O-linked glycosylation in ARPE-19 cells. Biomed Res Int 2014; 2014:701395.
21. Kil WH, Kim SM, Lee JE, Park KS, Nam SJ. Anticancer effect of silibinin on the xenograft model using MDA-MB-468 breast cancer cells. Ann Surg Treat Res 2014; 87:167-173.
22. Ting H, Deep G, Agarwal R. Molecular mechanisms of silibinin-mediated cancer chemoprevention with major emphasis on prostate cancer. AAPS J 2013; 15:707-716.
23. Wang ZX, Lu BB, Wang H, Cheng ZX, Yin YM. MicroRNA-21 modulates chemosensitivity of breast cancer cells to doxorubicin by targeting PTEN. Arch Med Res 2011; 42:281-290.
24. Babashah S, Sadeghizadeh M, Hajifathali A,
Tavirani MR, Zomorod MS, Ghadiani M, et al. Targeting of the signal transducer Smo links microRNA-326 to the oncogenic Hedgehog pathway in CD34+ CML stem/progenitor cells. Int J Cancer 2013; 133:579-589.
25. Yan LX, Huang XF, Shao Q, Huang MY, Deng L, Wu QL, et al. MicroRNA miR-21 overexpression in human breast cancer is associated with advanced clinical stage, lymph node metastasis and patient poor prognosis. RNA 2008; 14:2348-2360.
26. Iorio MV, Ferracin M, Liu CG, Veronese A, Spizzo R, Sabbioni S, et al. MicroRNA gene expression deregulation in human breast cancer. Cancer Res 2005; 65:7065-7070.
27. Jiang S, Zhang LF, Zhang HW, Hu S, Lu MH, Liang S, et al. A novel miR-155/miR-143 cascade controls glycolysis by regulating hexokinase 2 in breast cancer cells. EMBO J 2012; 31:1985-1998.
28. Gan R, Yang Y, Yang X, Zhao L, Lu J, Meng QH. Downregulation of miR-221/222 enhances sensitivity of breast cancer cells to tamoxifen through upregulation of TIMP3. Cancer Gene Ther 2014; 21:290-296.
29. Chen WX, Hu Q, Qiu MT, Zhong SL, Xu JJ, Tang JH, et al. miR-221/222: promising biomarkers for breast cancer. Tumour Biol 2013; 34:1361-1370.
30. Andorfer CA, Necela BM, Thompson EA, Perez EA. MicroRNA signatures: clinical biomarkers for the diagnosis and treatment of breast cancer. Trends Mol Med 2011; 17:313-319.
31. Negrini M, Nicoloso MS, Calin GA. MicroRNAs and cancer--new paradigms in molecular oncology. Curr Opin Cell Biol 2009; 21:470-479.
32. Selcuklu SD, Donoghue MT, Spillane C. miR-21 as a key regulator of oncogenic processes. Biochem Soc Trans 2009; 37:918-925.
33. Mattiske S, Suetani RJ, Neilsen PM, Callen DF. The oncogenic role of miR-155 in breast cancer. Cancer Epidemiol Biomarkers Prev 2012; 21:1236-1243.
34. Zhang CM, Zhao J, Deng HY. MiR-155 promotes proliferation of human breast cancer MCF-7 cells through targeting tumor protein 53-induced nuclear protein 1. J Biomed Sci 2013; 20:79.
35. Ekimler S, Sahin K. Computational methods for microRNA target prediction. Genes (Basel) 2014; 5:671-683.
36. Frankel LB, Christoffersen NR, Jacobsen A, Lindow M, Krogh A, Lund AH. Programmed cell death 4 (PDCD4) is an important functional target of the microRNA miR-21 in breast cancer cells. J Biol Chem 2008; 283:1026-1033.
37. Higgs G, Slack F. The multiple roles of microRNA-155 in oncogenesis. J Clin Bioinforma 2013; 3:17.
38. Fu WN, Bertoni F, Kelsey SM, McElwaine SM, Cotter FE, Newland AC, et al. Role of DNA methylation in the suppression of Apaf-1 protein in human leukaemia. Oncogene 2003; 22:451-455.
39. Kamarajan P, Sun NK, Sun CL, Chao CC. Apaf-1 overexpression partially overcomes apoptotic resistance in a cisplatin-selected HeLa cell line. FEBS Lett 2001; 505:206-212.
Iranian Journal of Basic Medical Sciences
Volume 18, Issue 12 - Serial Number 12
December 2015
Pages 1209-1214

Files

Share

How to cite

Statistics