Evaluation of the effects of UCA1 gene knockout with a new CRISPR/Cas9 gene editing technique in ovarian cancer cell line
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 kno...
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Jahrom University of Medical Sciences
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doaj-124d8b0561be4deda51fd4da77a335f42021-10-02T06:11:34ZfasJahrom University of Medical Sciencesفصلنامه دانشگاه علوم پزشکی جهرم2008-79932008-84422021-03-011911020Evaluation of the effects of UCA1 gene knockout with a new CRISPR/Cas9 gene editing technique in ovarian cancer cell lineBehshad Montazeri-Najafabadi0Abbas Doosti1Jafar Kiani2 Department of Biology, Faculty of Basic Sciences, Shahrekord Branch, Islamic Azad University, Shahrekord, Iran Biotechnology Research Center, Shahrekord Branch, Islamic Azad University, Shahrekord, Iran Department of Molecular Medicine, Faculty of Advanced Technologies in Medicine, Iran University of Medical Sciences (IUMS), Tehran, Iran 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.http://jmj.jums.ac.ir/article-1-1582-en.htmluca1ovarian cancercrispr/cas9 |
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language |
fas |
format |
Article |
sources |
DOAJ |
author |
Behshad Montazeri-Najafabadi Abbas Doosti Jafar Kiani |
spellingShingle |
Behshad Montazeri-Najafabadi Abbas Doosti Jafar Kiani Evaluation of the effects of UCA1 gene knockout with a new CRISPR/Cas9 gene editing technique in ovarian cancer cell line فصلنامه دانشگاه علوم پزشکی جهرم uca1 ovarian cancer crispr/cas9 |
author_facet |
Behshad Montazeri-Najafabadi Abbas Doosti Jafar Kiani |
author_sort |
Behshad Montazeri-Najafabadi |
title |
Evaluation of the effects of UCA1 gene knockout with a new CRISPR/Cas9 gene editing technique in ovarian cancer cell line |
title_short |
Evaluation of the effects of UCA1 gene knockout with a new CRISPR/Cas9 gene editing technique in ovarian cancer cell line |
title_full |
Evaluation of the effects of UCA1 gene knockout with a new CRISPR/Cas9 gene editing technique in ovarian cancer cell line |
title_fullStr |
Evaluation of the effects of UCA1 gene knockout with a new CRISPR/Cas9 gene editing technique in ovarian cancer cell line |
title_full_unstemmed |
Evaluation of the effects of UCA1 gene knockout with a new CRISPR/Cas9 gene editing technique in ovarian cancer cell line |
title_sort |
evaluation of the effects of uca1 gene knockout with a new crispr/cas9 gene editing technique in ovarian cancer cell line |
publisher |
Jahrom University of Medical Sciences |
series |
فصلنامه دانشگاه علوم پزشکی جهرم |
issn |
2008-7993 2008-8442 |
publishDate |
2021-03-01 |
description |
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. |
topic |
uca1 ovarian cancer crispr/cas9 |
url |
http://jmj.jums.ac.ir/article-1-1582-en.html |
work_keys_str_mv |
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