Long non-coding RNA UCA1 Knockdown Assisted by CRISPR/Cas9 in Female Cancer Cell Lines Increases Mir-143 Tumor-Suppressor.

IF 1.3 4区医学 Q4 PUBLIC, ENVIRONMENTAL & OCCUPATIONAL HEALTH

Iranian Journal of Public Health Pub Date : 2024-04-01 DOI:10.18502/ijph.v53i4.15571

Behshad Montazeri-Najafabadi, Abbas Doosti, Jafar Kiani

{"title":"Long non-coding RNA UCA1 Knockdown Assisted by CRISPR/Cas9 in Female Cancer Cell Lines Increases Mir-143 Tumor-Suppressor.","authors":"Behshad Montazeri-Najafabadi, Abbas Doosti, Jafar Kiani","doi":"10.18502/ijph.v53i4.15571","DOIUrl":null,"url":null,"abstract":"Background: The lncRNAs has been linked to several malignancies, including breast cancer. Our objective was to investigate the impact of urothelial carcinoma associated 1 (UCA1) on cellular growth and death by a CRISPR/Cas9 knockdown technique.Methods: In 2020, the CHOPCHOP program was utilized to design two sgRNAs targeting the UCA gene. sgRNA1 and sgRNA2 were inserted into two different CRISPR plasmids to produce two recombinant plasmids. These recombinant plasmids were simultaneously transfected into MCF-7 and MDA-MB 231 carcinoma of the breast cells. Proliferation and apoptosis were compared using the MTT test, CCK-8 assay, and flow cytometry evaluation. RNA-hybrid software, quantitative reverse transcription PCR, and luciferase assays were utilized to confirm the relationship between UCA1 and miR-143.Results: Proliferated cells were less active in MTT and CCK-8 tests and fellow cytometry analysis. The PX459-sgRNA1,2 group had elevated levels of the cancer biomarker Caspase-3 gene expression (P<0.001). When WT-UCA1 and miR-143 were co-transfected, the luciferase activity was drastically decreased.Conclusion: One very effective method of regulating cellular proliferation in vitro is the deletion of UCA1, which CRISPR/Cas9 accomplishes.","PeriodicalId":49173,"journal":{"name":"Iranian Journal of Public Health","volume":null,"pages":null},"PeriodicalIF":1.3000,"publicationDate":"2024-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11493584/pdf/","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Iranian Journal of Public Health","FirstCategoryId":"3","ListUrlMain":"https://doi.org/10.18502/ijph.v53i4.15571","RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q4","JCRName":"PUBLIC, ENVIRONMENTAL & OCCUPATIONAL HEALTH","Score":null,"Total":0}

引用次数: 0

Abstract

Background: The lncRNAs has been linked to several malignancies, including breast cancer. Our objective was to investigate the impact of urothelial carcinoma associated 1 (UCA1) on cellular growth and death by a CRISPR/Cas9 knockdown technique.

Methods: In 2020, the CHOPCHOP program was utilized to design two sgRNAs targeting the UCA gene. sgRNA1 and sgRNA2 were inserted into two different CRISPR plasmids to produce two recombinant plasmids. These recombinant plasmids were simultaneously transfected into MCF-7 and MDA-MB 231 carcinoma of the breast cells. Proliferation and apoptosis were compared using the MTT test, CCK-8 assay, and flow cytometry evaluation. RNA-hybrid software, quantitative reverse transcription PCR, and luciferase assays were utilized to confirm the relationship between UCA1 and miR-143.

Results: Proliferated cells were less active in MTT and CCK-8 tests and fellow cytometry analysis. The PX459-sgRNA1,2 group had elevated levels of the cancer biomarker Caspase-3 gene expression (P<0.001). When WT-UCA1 and miR-143 were co-transfected, the luciferase activity was drastically decreased.

Conclusion: One very effective method of regulating cellular proliferation in vitro is the deletion of UCA1, which CRISPR/Cas9 accomplishes.

查看原文本刊更多论文

通过 CRISPR/Cas9 在雌性癌症细胞系中敲除长非编码 RNA UCA1 可增加 Mir-143 肿瘤抑制因子。

背景：lncRNAs与包括乳腺癌在内的多种恶性肿瘤有关。我们的目的是通过CRISPR/Cas9基因敲除技术研究尿道癌相关1（UCA1）对细胞生长和死亡的影响：2020年，利用CHOPCHOP程序设计了两个靶向UCA基因的sgRNA，并将sgRNA1和sgRNA2插入到两个不同的CRISPR质粒中，产生了两个重组质粒。这些重组质粒被同时转染到 MCF-7 和 MDA-MB 231 乳腺癌细胞中。使用 MTT 试验、CCK-8 检测法和流式细胞仪评估比较细胞的增殖和凋亡。利用 RNA 杂交软件、定量反转录 PCR 和荧光素酶检测来确认 UCA1 和 miR-143 之间的关系：结果：增殖的细胞在 MTT 和 CCK-8 试验以及研究人员的细胞计数分析中活性较低。PX459-sgRNA1,2组癌症生物标志物Caspase-3基因表达水平升高（PUCA1和miR-143共转染后，荧光素酶活性急剧下降）：体外调节细胞增殖的一种非常有效的方法是删除 UCA1，而 CRISPR/Cas9 就能做到这一点。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

求助全文

约1分钟内获得全文求助全文

来源期刊

Iranian Journal of Public Health PUBLIC, ENVIRONMENTAL & OCCUPATIONAL HEALTH-

CiteScore

2.20

自引率

7.10%

发文量

300

审稿时长

3-8 weeks

期刊介绍： Iranian Journal of Public Health has been continuously published since 1971, as the only Journal in all health domains, with wide distribution (including WHO in Geneva and Cairo) in two languages (English and Persian). From 2001 issue, the Journal is published only in English language. During the last 41 years more than 2000 scientific research papers, results of health activities, surveys and services, have been published in this Journal. To meet the increasing demand of respected researchers, as of January 2012, the Journal is published monthly. I wish this will assist to promote the level of global knowledge. The main topics that the Journal would welcome are: Bioethics, Disaster and Health, Entomology, Epidemiology, Health and Environment, Health Economics, Health Services, Immunology, Medical Genetics, Mental Health, Microbiology, Nutrition and Food Safety, Occupational Health, Oral Health. We would be very delighted to receive your Original papers, Review Articles, Short communications, Case reports and Scientific Letters to the Editor on the above mentioned research areas.