Back
 IJCM  Vol.8 No.6 , June 2017
Doxorubicin Induces Apoptosis through down Regulation of miR-21 Expression and Increases miR-21 Target Gene Expression in MCF-7 Breast Cancer Cells
Abstract: miRNAs play an important regulatory role in variety of cellular functions and several diseases, including cancer. MicroRNA-21 (miR-21) is overexpressed in almost all types of human cancers. Studies revealed that the knockdown of miR-21 results in reduced tumor cell growth, cell cycle arrest and cell apoptosis. In this study, we evaluated the effect of doxorubicin on miR-21 expression in mcf-7 breast cancer cells. miRNA was extracted from mcf-7 cells treated with doxorubicin and untreated cells using miRNeasy Kit (Qiagen) according to the manufacturer’s instructions. cDNA synthesis was performed using miScript II RT Kit (Qiagen) and Real Time-PCR was performed using Real Q Plus 2x Master Mix Green-(Ampliqon, Denmark). The relative expression of miR-16 and miR-21 was calculated using comparative Ct method. All tests were run in triplicate to minimize the experimental errors. Samples with a Ct > 37 were excluded from the analysis. Statistically, a significant decrease in cell proliferation of mcf-7 cells was found in doxorubicin group compared with control groups 24 hours after transfection, dose dependently (p value< 0.001). After 24 hours, Doxorubicin (100 μm) significantly decreased miR-21 expression in mcf-7 cells (p = 0.0001). Also, the expression of caspase 9 significantly increased after Doxorubicin (100 μm) treatment (p = 0.0003). Together, these findings indicate that miR-21 plays a key role in regulating cell apoptosis in mcf-7 cells and may serve as a target for effective therapies.
Cite this paper: Tofigh, R. , Akhavan, S. , Tarban, N. , Sadrabadi, A. , Jalili, A. , Moridi, K. and Tutunchi, S. (2017) Doxorubicin Induces Apoptosis through down Regulation of miR-21 Expression and Increases miR-21 Target Gene Expression in MCF-7 Breast Cancer Cells. International Journal of Clinical Medicine, 8, 386-394. doi: 10.4236/ijcm.2017.86036.
References

[1]   Sara, T., Mojtaba, S., Mahta, M., Parisa, N., Bahareh, N., Reza, S. and Nasrin, G. (2016) How Does the Cortactin Gene Expression Affect Breast Cancer among Iranian Females? Advances in Breast Cancer Research, 5, 142-149.
https://doi.org/10.4236/abcr.2016.54017

[2]   Jalili, A., Roshandel, E., Shenasifam, S.H.F.N., Ebrahimi, A. and Sadrabadi, M.A.R.P. (2016) Appraising the Increased Expression of p21and p53 Genes in Paraffin-Embedded Biopsy Tissues of Patients with Bladder Cancer Exposed to Mitomycin C. Advances in Bioresearch, 7, 74-79.

[3]   Jalili, A., Sadrabadi, A.E. and Yekta, S.S. (2015) Assessing the Expression of BRAF Gene in Paraffin-Embedded Blocks of Patients with Colorectal Cancer. International Journal of Biology, Pharmacy and Allied Sciences, 4, 5653-5662.

[4]   Mohammadi, S., Mahboubi, A., Mohammadi, M., Hedayati, M. and Jalili, A. (2017) Assessing the Anticancer Effect of the Euphorbia Condylocarpa Plant on AGS Gastric Cancer Cell Line. Gene, Cell and Tissue, 4, e41223.
https://doi.org/10.17795/gct-41223

[5]   Sadrabadi, A.E., Yekta, S.S., Fattahi, S.H., Molaei, S., Gavgani, S.P., Roshandel, E. and Jalili, A. (2015) Assessing the Decreased Expression of TP53 and P21 in Colorectal Cancer. Indian Journal of Fundamental and Applied Life Sciences, 5, 107-112.

[6]   Ferreira, C.A., Fuscaldi, L.L., Townsend, D.M., Rubello, D., Barros, A.L. (2017) Radiolabeled Bombesin Derivatives for Preclinical Oncological Imaging. Biomedicine & Pharmacotherapy, 87, 58-72. https://doi.org/10.1016/j.biopha.2016.12.083

[7]   Huang, X., Xiao, R., Pan, S., Yang, X., Yuan, W., Tu, Z., et al. (2017) Uncovering the Roles of Long Non-Coding RNAs in Cancer Stem Cells. Journal of hematology & oncology, 10, 62.
https://doi.org/10.1186/s13045-017-0428-9

[8]   Elfandi, L., Said, G., Saleh, S.S., Marwan, M. and Enattah, N. (2016) Analysis of 6174delT Mutation in BRCA2 Gene by Mutagenically Separated PCR among Libyan Patients with Breast Cancer. Archives of Breast Cancer, 3, 8-13.

[9]   Lee, E. and Moon, A. (2016) Identification of Biomarkers for Breast Cancer Using Databases. Journal of Cancer Prevention, 21, 235-242.
https://doi.org/10.15430/JCP.2016.21.4.235

[10]   Chatterjee, S.K. and Zetter, B.R. (2005) Cancer Biomarkers: Knowing the Present and Predicting the Future. Future Oncology, 1, 37-50.
https://doi.org/10.1517/14796694.1.1.37

[11]   Tanic, M. and Beck, S. (2017) Epigenome-Wide Association Studies for Cancer Biomarker Discovery in Circulating Cell-Free DNA: Technical Advances and Challenges. Current Opinion in Genetics & Development, 42, 48-55.
https://doi.org/10.1016/j.gde.2017.01.017

[12]   Wang, H., Shi, T., Qian, W.-J., Liu, T., Kagan, J., Srivastava, S., et al. (2016) The Clinical Impact of Recent Advances in LC-MS for Cancer Biomarker Discovery and Verification. Expert Review of Proteomics, 13, 99-114.
https://doi.org/10.1586/14789450.2016.1122529

[13]   Li, Y., Deng, X., Zeng, X. and Peng, X. (2016) The Role of Mir-148a in Cancer. Journal of Cancer, 7, 1233-1241.
https://doi.org/10.7150/jca.14616

[14]   Denzler, R., McGeary, S.E., Title, A.C., Agarwal, V., Bartel, D.P. and Stoffel, M. (2016) Impact of MicroRNA Levels, Target-Site Complementarity, and Cooperativity on Competing Endogenous RNA-Regulated Gene Expression. Molecular Cell, 64, 565-579.
https://doi.org/10.1016/j.molcel.2016.09.027

[15]   Dykxhoorn, D.M. (2010) MicroRNAs and Metastasis: Little RNAs Go a Long Way. Cancer Research, 70, 6401-6406. https://doi.org/10.1158/0008-5472.CAN-10-1346

[16]   Jafri, M.A., Al-Qahtani, M.H. and Shay, J.W., Eds. (2017) Role of miRNAs in Human Cancer Metastasis: Implications for Therapeutic Intervention. Seminars in Cancer Biology, in press.

[17]   Livingstone, M.C., Johnson, N.M., Roebuck, B.D., Kensler, T.W. and Groopman, J.D. (2017) Profound Changes in miRNA Expression during Cancer Initiation by Aflatoxin B1 and Their Abrogation by the Chemopreventive Triterpenoid CDDO-Im. Molecular Carcinogenesis, in press.
https://doi.org/10.1002/mc.22635

[18]   Singh, S., Sharma, P.K., Kumar, N. and Dudhe, R. (2011) A Review on a Versatile Molecule: Chalcone. Asian J Pharm Biol Res, 412-418.

[19]   Klinge, C.M. (2015) miRNAs Regulated by Estrogens, Tamoxifen, and Endocrine Disruptors and Their Downstream Gene Targets. Molecular and Cellular Endocrinology, 418, 273-297.
https://doi.org/10.1016/j.mce.2015.01.035

[20]   Wang, S., Wan, X. and Ruan, Q. (2016) The MicroRNA-21 in Autoimmune Diseases. International Journal of Molecular Sciences, 17, 864.
https://doi.org/10.3390/ijms17060864

[21]   Volinia, S., Galasso, M., Costinean, S., Tagliavini, L., Gamberoni, G., Drusco, A., et al. (2010) Reprogramming of miRNA Networks in Cancer and Leukemia. Genome Research, 20, 589-599.
https://doi.org/10.1101/gr.098046.109

[22]   Croce, C.M. (2011) miRNAs in the Spotlight: Understanding Cancer Gene Dependency. Nature Medicine, 17, 935-936.
https://doi.org/10.1038/nm0811-935

[23]   Papagiannakopoulos, T., Shapiro, A. and Kosik, K.S. (2008) MicroRNA-21 Targets a Network of Key Tumor-Suppressive Pathways in Glioblastoma Cells. Cancer Research, 68, 8164-8172.
https://doi.org/10.1158/0008-5472.CAN-08-1305

[24]   Park, J.-K., Lee, E.J., Esau, C. and Schmittgen, T.D. (2009) Antisense Inhibition of microRNA-21 or-221 Arrests Cell Cycle, Induces Apoptosis, and Sensitizes the Effects of Gemcitabine in Pancreatic Adenocarcinoma. Pancreas, 38, e190-e199.
https://doi.org/10.1097/MPA.0b013e3181ba82e1

[25]   Tomuleasa, C.I., Foris, V., Soritau, O., Pall, E., Fischer-Fodor, E., Lung-Illes, V., et al. (2009) Effects of 60Co Gamma-Rays on Human Osteoprogenitor Cells. Romanian Journal of Morphology and Embryology, 50, 349-355.

[26]   Cramer, H., Lauche, R., Klose, P., Lange, S., Langhorst, J. and Dobos, G.J. (2017) Yoga for Improving Health-Related Quality of Life, Mental Health and Cancer-Related Symptoms in Women Diagnosed with Breast Cancer. The Cochrane Database of Systematic Reviews, No. 1, CD010802.
https://doi.org/10.1002/14651858.cd010802.pub2

[27]   Lobo, M.D., Moreno, F.B., Souza, G.H., Verde, S.M., Moreira, R.A. and Monteiro-Moreira, A.C. (2017) Label-Free Proteome Analysis of Plasma from Patients with Breast Cancer: Stage-Specific Protein Expression. Frontiers in Oncology, 7, 14.
https://doi.org/10.3389/fonc.2017.00014

[28]   Owen, L.N. (1979) A Comparative Study of Canine and Human Breast Cancer. Investigative & Cell Pathology, 2, 257-275.

[29]   Arslan, C., Dizdar, O. and Altundag, K. (2014) Chemotherapy and Biological Treatment Options in Breast Cancer Patients with Brain Metastasis: An Update. Expert Opinion on Pharmacotherapy, 15, 1643-1658.
https://doi.org/10.1517/14656566.2014.929664

[30]   Jolly, T.A., Williams, G.R., Bushan, S., Pergolotti, M., Nyrop, K.A., Jones, E.L., et al. (2016) Adjuvant Treatment for Older Women with Invasive Breast Cancer. Women’s Health (London, England), 12, 129-145; quiz 45-46.

[31]   Pal, A. and Donato, N.J. (2014) Ubiquitin-Specific Proteases as Therapeutic Targets for the Treatment of Breast Cancer. Breast Cancer Research, 16, 461.
https://doi.org/10.1186/s13058-014-0461-3

[32]   Casalini, P. and Iorio, M.V. (2009) MicroRNAs and Future Therapeutic Applications in Cancer. Journal of BUON, 14, S17-S22.

[33]   Johnson, S.M., Grosshans, H., Shingara, J., Byrom, M., Jarvis, R., Cheng, A., et al. (2005) RAS Is Regulated by the let-7 microRNA Family. Cell, 120, 635-647.
https://doi.org/10.1016/j.cell.2005.01.014

[34]   Croce, C.M. (2009) Causes and Consequences of microRNA Dysregulation in Cancer. Nature Reviews Genetics, 10, 704-714. https://doi.org/10.1038/nrg2634

[35]   Volinia, S., Calin, G.A., Liu, C.G., Ambs, S., Cimmino, A., Petrocca, F., et al. (2006) A microRNA Expression Signature of Human Solid Tumors Defines Cancer Gene Targets. Proceedings of the National Academy of Sciences of the United States of America, 103, 2257-2261.
https://doi.org/10.1073/pnas.0510565103

[36]   Si, M.L., Zhu, S., Wu, H., Lu, Z., Wu, F. and Mo, Y.Y. (2007) miR-21-Mediated Tumor Growth. Oncogene, 26, 2799-2803.
https://doi.org/10.1038/sj.onc.1210083

[37]   Hwang, J.H., Voortman, J., Giovannetti, E., Steinberg, S.M., Leon, L.G., Kim, Y.T., et al. (2010) Identification of microRNA-21 as a Biomarker for Chemoresistance and Clinical Outcome Following Adjuvant Therapy in Resectable Pancreatic Cancer. PLoS ONE, 5, e10630.
https://doi.org/10.1371/journal.pone.0010630

[38]   Yan, L.-X., Huang, X.-F., Shao, Q., Huang, M.A.Y., Deng, L., Wu, Q.-L., et al. (2008) MicroRNA miR-21 Overex-pression in Human Breast Cancer Is Associated with Advanced Clinical Stage, Lymph Node Metastasis and Patient Poor Prognosis. RNA, 14, 2348-2360.
https://doi.org/10.1261/rna.1034808

[39]   Buscaglia, L.E.B. and Li, Y. (2011) Apoptosis and the Target Genes of microRNA-21. Chinese Journal of Cancer, 30, 371-380.
https://doi.org/10.5732/cjc.30.0371

[40]   Yan, L.X., Wu, Q.N., Zhang, Y., Li, Y.Y., Liao, D.Z., Hou, J.H., et al. (2011) Knockdown of miR-21 in Human Breast Cancer Cell Lines Inhibits Proliferation, in Vitro Migration and in Vivo Tumor Growth. Breast Cancer Research, 13, R2.

 
 
Top