Evaluation of the effects of UCA1 gene knockout with a new CRISPR/Cas9 gene editing technique in ovarian cancer cell line

Authors
Behshad Montazeri-Najafabadi 1
Abbas Doosti 2
Jafar Kiani 3
1 Department of Biology, Faculty of Basic Sciences, Shahrekord Branch, Islamic Azad University, Shahrekord, Iran
2 Biotechnology Research Center, Shahrekord Branch, Islamic Azad University, Shahrekord, Iran
3 Department of Molecular Medicine, Faculty of Advanced Technologies in Medicine, Iran University of Medical Sciences (IUMS), Tehran, Iran
10.52547/jmj.19.1.3
Abstract
Introduction: lncRNAs play their roles in cell growth, transcription, translation, and apoptosis. UCA1 is a type of lncRNA has a direct relationship with ovarian cancer. More than half of the women with ovarian cancer die within less than five years from the disease diagnosis. This study aims to knockout of UCA1 gene using the CRISPR/Cas9 technique in SK-OV-3 cell and studying its effect on cell factors involved in cancer progress and apoptosis.
Materials and Methods: Two types of sgRNAs for UCA1 gene were designed. sgRNAs were cloned after synthesis in two
vectors of PX459. The SK-OV-3 cell was transfected with two vectors carrying sgRNAs. The expression of FAS, BAK, BAX, and P53 pro-apoptosis genes and BCL2 and SURVIVIN anti-apoptosis genes were evaluated using the q-PCR. Besides, the cell proliferation rate and apoptosis induction were studied using MTT and flowcytometry.
Results: The result showed the successful knockout of the UCA1 gene in the SK-OV-3 cell line. The expression of FAS, BAK, BAX, and P53 genes showed a meaningful increase related to the expression in manipulated cells compared to control cells (p˂0.05). Also, the expression decrease of BCL2 and SURVIVIN genes was seen (p˂0.05). MTT and flow cytometry results confirmed the cell proliferation decrease and the apoptosis increase.
Conclusion: UCA1 gene Knockout in the SK-OV-3 showed some positive effects in decreasing cancer cell proliferation and increasing the apoptosis rate can be said that inhibiting overexpression of the UCA1 gene will play a positive role in controlling the growth of ovarian cancer cells.
Keywords
UCA1
Ovarian Cancer
CRISPR/CAS9
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