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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Main Authors: Behshad Montazeri-Najafabadi, Abbas Doosti, Jafar Kiani
Format: Article
Language:fas
Published: Jahrom University of Medical Sciences 2021-03-01
Series:فصلنامه دانشگاه علوم پزشکی جهرم
Subjects:
Online Access:http://jmj.jums.ac.ir/article-1-1582-en.html
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spelling 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
collection DOAJ
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
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AT jafarkiani evaluationoftheeffectsofuca1geneknockoutwithanewcrisprcas9geneeditingtechniqueinovariancancercellline
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