Differential Expression of EGFR, MAP2K4 and E2F3 Genes as Targets of miR-141 and Its Association With Immune System Pathway

Document Type : Research Articles

Authors

• Soheila shokrollahzade ¹
• Arshad Hosseini ¹
• Majid Safa ¹
• Shamim Sarhadi ²

¹ Iran University of Medical Sciences

² Tabriz University of Medical Sciences, Tabriz, Iran

https://doi.org/10.22067/jcmr.v0i0.58965

Abstract

MicroRNAs by their structural complementarity capabilities have canonical roles in gene regulation. In this paper; we investigate expression of EGFR, MAP2K4 and E2F3 genes targeted by miR-141, a member of miR-200 family. EGFR, MAP2K4 and E2F3 were predicted as the potential targets of mir-141 by using online miRNA bioinformatics tools. MCF-7 cells were transfected with mir-141-precursor and inhibitor vectors. Expression of miR-141 and target genes was determined by using qRT-PCR. To see the most relevant pathways regulated by miR-141, we constructed two separate networks by NetworkAnalyst and enriched list of underlying genes by Enrichment analysis tools. The expression changes of all three predicted targets were higher in transfected cells with anti-mir-141 vector, compared with the control untransfected cells. By contrast, in transfected cells with pre-mir 141, we did not see significant expression changes in EGFR, E2F3 and MAP2K4. List of genes in total networks as well as explored functional modules were enriched separately. Enrichment analysis shows that immune system pathway has the strongest relationship with the proteins potentially targeted by miR-141. The present study demonstrated potential role of miR-141 in regulation of EGFR, MAP2K4 and E2F3 expression and suggested innate immunity pathways as the key pathway through which this regulatory network contributes to breast cancer development.

Keywords

• Key Words: Breast cancer
• miR-141
• EGFR
• MAP2K4
• E2F3

20.1001.1.20089147.2017.9.1.3.8

References

1.Antolin S., Calvo L., Blanco-Calvo M., Santiago M. P., Lorenzo-Patino M. J., Haz-Conde M., Santamarina I., Figueroa A., Anton-Aparicio L. M. and Valladares-Ayerbes M. (2015) Circulating miR-200c and miR-141 and outcomes in patients with breast cancer. BMC Cancer 15:297.

2.Canturk K. M., Ozdemir M., Can C., Oner S., Emre R., Aslan H., Cilingir O., Ciftci E., Celayir F. M., Aldemir O., Ozen M. and Artan S. (2014) Investigation of key miRNAs and target genes in bladder cancer using miRNA profiling and bioinformatic tools. Molecular biology reports 41:8127-8135.

3.Chen L., Gibbons D. L., Goswami S., Cortez M. A., Ahn Y. H., Byers L. A., Zhang X., Yi X., Dwyer D., Lin W., Diao L., Wang J., Roybal J. D., Patel M., Ungewiss C., Peng D., Antonia S., Mediavilla-Varela M., Robertson G., Jones S., Suraokar M., Welsh J. W., Erez B., Wistuba, II, Chen L., Peng D., Wang S., Ullrich S. E., Heymach J. V., Kurie J. M. and Qin F. X. (2014) Metastasis is regulated via microRNA-200/ZEB1 axis control of tumour cell PD-L1 expression and intratumoral immunosuppression. Nat Commun 5:5241.

4.Gazdar A. F. (0000) Activating and resistance mutations of EGFR in non-small-cell lung cancer: role in clinical response to EGFR tyrosine kinase inhibitors. Oncogene 28:S24-S31.

5.Kim Y. K., Yeo J., Kim B., Ha M. and Kim V. N. (2012) Short structured RNAs with low GC content are selectively lost during extraction from a small number of cells. Molecular cell 46:893-895.

6.Luo D., Wilson J. M., Harvel N., Liu J., Pei L., Huang S., Hawthorn L. and Shi H. (2013) A systematic evaluation of miRNA:mRNA interactions involved in the migration and invasion of breast cancer cells. Journal of translational medicine 11:1-14.

7.Marasa B. S., Srikantan S., Masuda K., Abdelmohsen K., Kuwano Y., Yang X., Martindale J. L., Rinker-Schaeffer C. W. and Gorospe M. (2009) Increased MKK4 abundance with replicative senescence is linked to the joint reduction of multiple microRNAs. Science signaling 2:ra69.

8.Masuda H., Zhang D., Bartholomeusz C., Doihara H., Hortobagyi G. N. and Ueno N. T. (2012) Role of epidermal growth factor receptor in breast cancer. Breast cancer research and treatment 136:331-345.

9.Naghavian R., Ghaedi K., Kiani-Esfahani A., Ganjalikhani-Hakemi M., Etemadifar M. and Nasr-Esfahani M. H. (2015) miR-141 and miR-200a, Revelation of New Possible Players in Modulation of Th17/Treg Differentiation and Pathogenesis of Multiple Sclerosis. PLoS One 10:e0124555.

10.Neves R., Scheel C., Weinhold S., Honisch E., Iwaniuk K. M., Trompeter H.-I., Niederacher D., Wernet P., Santourlidis S. and Uhrberg M. (2010) Role of DNA methylation in miR-200c/141 cluster silencing in invasive breast cancer cells. BMC Research Notes 3:1-7.

11.Nguyen-Vu T., Vedin L. L., Liu K., Jonsson P., Lin J. Z., Candelaria N. R., Candelaria L. P., Addanki S., Williams C., Gustafsson J. A., Steffensen K. R. and Lin C. Y. (2013) Liver x receptor ligands disrupt breast cancer cell proliferation through an E2F-mediated mechanism. Breast cancer research : BCR 15:R51.

12.Park S. M., Gaur A. B., Lengyel E. and Peter M. E. (2008) The miR-200 family determines the epithelial phenotype of cancer cells by targeting the E-cadherin repressors ZEB1 and ZEB2. Genes & development 22:894-907.

13.Rusek A. M., Abba M., Eljaszewicz A., Moniuszko M., Niklinski J. and Allgayer H. (2015) MicroRNA modulators of epigenetic regulation, the tumor microenvironment and the immune system in lung cancer. Molecular Cancer 14:34.

14.Shimono Y., Zabala M., Cho R. W., Lobo N., Dalerba P., Qian D., Diehn M., Liu H., Panula S. P., Chiao E., Dirbas F. M., Somlo G., Pera R. A., Lao K. and Clarke M. F. (2009) Downregulation of miRNA-200c links breast cancer stem cells with normal stem cells. Cell 138:592-603.

15.Taylor M. A. and Schiemann W. P. (2014) Therapeutic opportunities for targeting microRNAs in cancer. Molecular and Cellular Therapies 2:1-13.

16.Ueda R., Kohanbash G., Sasaki K., Fujita M., Zhu X., Kastenhuber E. R., McDonald H. A., Potter D. M., Hamilton R. L., Lotze M. T., Khan S. A., Sobol R. W. and Okada H. (2009) Dicer-regulated microRNAs 222 and 339 promote resistance of cancer cells to cytotoxic T-lymphocytes by down-regulation of ICAM-1. Proceedings of the National Academy of Sciences 106:10746-10751.

17.Uhlmann S., Zhang J. D., Schwager A., Mannsperger H., Riazalhosseini Y., Burmester S., Ward A., Korf U., Wiemann S. and Sahin O. (2010) miR-200bc/429 cluster targets PLCgamma1 and differentially regulates proliferation and EGF-driven invasion than miR-200a/141 in breast cancer. Oncogene 29:4297-4306.

18.Wang L., Pan Y. and Dai J. L. (2004) Evidence of MKK4 pro-oncogenic activity in breast and pancreatic tumors. Oncogene 23.

19.Xu L., Yang B. F. and Ai J. (2013) MicroRNA transport: a new way in cell communication. Journal of cellular physiology 228:1713-1719.

20.Xue J., Niu Y. F., Huang J., Peng G., Wang L. X., Yang Y. H. and Li Y. Q. (2014) miR-141 suppresses the growth and metastasis of HCC cells by targeting E2F3. Tumour biology : the journal of the International Society for Oncodevelopmental Biology and Medicine 35:12103-12107.