Role of mir-214 in modulating chemoresistance and its effect on global mRNA expression profiling in cervical cancer


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Prakriti Sen1,Niladri Ganguly2
1KIIT School Of Biotechnology,2KIIT School of Biotechnology

Abstract

Background

Cervical carcinoma is the second most prevalent cancer and the fourth most common reason of death due to cancer in women. In cervical cancer, there are about 250 different micro RNAs (mi-RNAs) have been found to be deregulated. In this study we investigated the effect of tumour suppressor mi-RNA, miR-214 in modulating chemo-resistance. We also investigated the global mRNA expression profile by RNA-seq after overexpression and knockdown of miR-214 in cervical cancer cells.

Method

miR-214 was knocked-out and over-expressed in different cervical cancer cell lines. Post and pre-transfection cervical cancer cells were exposed to doxorubicin, cisplatin and paclitaxel and their role in chemoresistance and PI-3Kinase/Akt signalling pathway were investigated. Furthermore, the global mRNA expression profile was performed in miR-214 knockdown and overexpressed cervical cancer cells by using Illumina Hiseq 2500. Gene Ontology (GO) and KEGG pathway analysis were performed to identify the differentially expressed genes (DEG) followed by network analysis.

Results

We observed that knock-out of miR-214 resulted in reduced apoptosis and exhibited increased chemoresistance while over expression of miR-214 exhibited chemosensitizes cervical cancer cells towards chemotherapeutic drugs. We observed that paclitaxel was most effective in inducing cell death. Subsequent analysis of apoptotic markers by western blot analysis showed that miR-214 over expression along with paclitaxel treatment caused an increase in expression of PARP and decline of PI-3 kinase/Akt levels. Global mRNA expression profiling of miR-214 overexpressed and knock-out cervical cancer cells exhibited differential gene expression. GO and KEGG pathway analysis followed by network analysis revealed distinct biological processes and pathways being triggered in miR-214 overexpressed and knock-out cells.

Conclusion

In conclusion, we observed that combination of miR-214 and paclitaxel treatment chemosensitizes cervical cancer cells. RNA Seq analysis revealed the differential effect of miR-214 on biological responses based on its expression level in cervical cancer.