CRISPR-mediated knockdown of miR-214 modulates cell fate in response to anti-cancer drugs in HPV-negative and HPVpositive cervical cancer cells
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Date
2020-06
Journal Title
Journal ISSN
Volume Title
Publisher
Indian Academy of Sciences
Abstract
Cervical cancer is the fourth most common cause of mortality in women worldwide. In this study weinvestigated the effect of a tumour suppressor microRNA miR-214 in modulating the cell death againstchemotherapeutic drugs like Doxorubicin, Cisplatin and Paclitaxel. CRISPR-facilitated knockdown andplasmid-based overexpression of miR-214 was performed in cervical cancer cell lines HeLa, C33A and CaSki.It was observed that knocking out miR-214 resulted in reduced apoptosis and cell migration upon drugtreatments; while overexpression of miR-214 resulted in marginal increase in apoptosis and cell migrationwhen treated with drugs. However, miR-214 had very little effect on production of reactive oxygen species.Our results also indicate that Doxorubicin was least effective and Paclitaxel most effective in inducing celldeath. A combination of miR-214 overexpression and Paclitaxel treatment was found to be most effective ininducing cell death in cervical cancer cells. Analysis of cell cycle phases followed by apoptotic markers alsoshowed that miR-214 overexpression along with Paclitaxel treatment caused an increase in PARP and declineof PI-3 kinase/Akt levels. Therefore, miR-214 levels determine the fate of the cancer cell duringchemotherapeutic treatment.
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Keywords
Cervical cancer, chemotherapy, CRISPR knockdown, HPV-positive and HPV-negative
Citation
Sen Prakriti, Ghosal Sayam, Hazra Rudranil, Mohanty Rimjhim, Arega Solomon, Sahu Bikash, Ganguly Niladri. CRISPR-mediated knockdown of miR-214 modulates cell fate in response to anti-cancer drugs in HPV-negative and HPVpositive cervical cancer cells. Journal of Biosciences. 2020 Jun; : 1-17