CRISPR/Cas9, a new approach to successful knockdown of ABCB1/P-glycoprotein and reversal of chemosensitivity in human epithelial ovarian cancer cell line

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Norouzi Barough Leyla; Sarookhani Mohammadreza; SALEHI RASOUL; SHARIFI MOHAMMADREZA; MOGHBELINEJAD SAHAR

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Journal Paper

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Journal: Iranian Journal of Basic Medical Sciences Year:2018 | Volume:21 | Issue:2 Page(s): 181-187

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Title

CRISPR/Cas9, a new approach to successful knockdown of ABCB1/P-glycoprotein and reversal of chemosensitivity in human epithelial ovarian cancer cell line

Author(s)

Keywords

CRISPR

Doxorubicin

Drug resistance

Ovarian cancer

P-glycoprotein

Abstract

Objective(s): MultiDrug resistance (MDR) is a major obstacle in the successful chemotherapy of Ovarian cancer. Inhibition of P-glycoprotein (P-gp), a member of ATP-binding cassette (ABC) transporters, is a well-known strategy to overcome MDR in cancer. The aim of this study was to investigate the efficiency and ability of CRISPR/Cas9 genome editing technology to knockdown ABCB1 gene expression in adriamycin resistant (A2780/ADR) Ovarian cancer cell line and evaluate the sensitivity changes to Doxorubicin. Materials and Methods: Three single-guide RNAs (sgRNAs) targeting the fourth and fifth exons of human ABCB1 gene were designed in this study. Expression level of ABCB1 was detected using quantitative real time PCR (qRT-PCR) after co-transfection of all three sgRNAs into A2780/ADR cell line and subsequent antibiotic selection. Drug sensitivity to Doxorubicin was determined by the 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) assay. Results: The results showed that CRISPR/Cas9 system could significantly reduce the expression of P-gp. The dramatic decline in ABCB1 gene expression was associated with increased sensitivity of cells transfected with sgRNAs to Doxorubicin. Conclusion: Based on the results of this study, it is concluded that the CRISPR-based systems, used in the present study, effectively down-regulated the target gene and acted as an ideal and cost-effective tool for gene editing of A2780/ADR cell line resulting in restoration of nonmalignant phenotype.

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APA: Copy

Norouzi Barough, Leyla, Sarookhani, Mohammadreza, SALEHI, RASOUL, SHARIFI, MOHAMMADREZA, & MOGHBELINEJAD, SAHAR. (2018). CRISPR/Cas9, a new approach to successful knockdown of ABCB1/P-glycoprotein and reversal of chemosensitivity in human epithelial ovarian cancer cell line. IRANIAN JOURNAL OF BASIC MEDICAL SCIENCES, 21(2), 181-187. SID. https://sid.ir/paper/721876/en

Vancouver: Copy

Norouzi Barough Leyla, Sarookhani Mohammadreza, SALEHI RASOUL, SHARIFI MOHAMMADREZA, MOGHBELINEJAD SAHAR. CRISPR/Cas9, a new approach to successful knockdown of ABCB1/P-glycoprotein and reversal of chemosensitivity in human epithelial ovarian cancer cell line. IRANIAN JOURNAL OF BASIC MEDICAL SCIENCES[Internet]. 2018;21(2):181-187. Available from: https://sid.ir/paper/721876/en

IEEE: Copy

Leyla Norouzi Barough, Mohammadreza Sarookhani, RASOUL SALEHI, MOHAMMADREZA SHARIFI, and SAHAR MOGHBELINEJAD, “CRISPR/Cas9, a new approach to successful knockdown of ABCB1/P-glycoprotein and reversal of chemosensitivity in human epithelial ovarian cancer cell line,” IRANIAN JOURNAL OF BASIC MEDICAL SCIENCES, vol. 21, no. 2, pp. 181–187, 2018, [Online]. Available: https://sid.ir/paper/721876/en

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