Behshad Montazeri-Najafabadi, Abbas Doosti, Jafar Kiani
{"title":"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。","authors":"Behshad Montazeri-Najafabadi, Abbas Doosti, Jafar Kiani","doi":"10.18502/ijph.v53i4.15571","DOIUrl":null,"url":null,"abstract":"<p><strong>Background: </strong>The lncRNAs has been linked to several malignancies, including breast cancer. Our objective was to investigate the impact of urothelial carcinoma associated 1 (<i>UCA1</i>) on cellular growth and death by a CRISPR/Cas9 knockdown technique.</p><p><strong>Methods: </strong>In 2020, the CHOPCHOP program was utilized to design two sgRNAs targeting the <i>UCA ge</i>ne. 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 <i>UCA1</i> and miR-143.</p><p><strong>Results: </strong>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 <i>Caspase-3</i> gene expression (<i>P</i><0.001). When WT-<i>UCA1</i> and miR-143 were co-transfected, the luciferase activity was drastically decreased.</p><p><strong>Conclusion: </strong>One very effective method of regulating cellular proliferation in vitro is the deletion of <i>UCA1</i>, which CRISPR/Cas9 accomplishes.</p>","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":"{\"title\":\"Long non-coding RNA <i>UCA1</i> Knockdown Assisted by CRISPR/Cas9 in Female Cancer Cell Lines Increases Mir-143 <i>Tumor-Suppressor</i>.\",\"authors\":\"Behshad Montazeri-Najafabadi, Abbas Doosti, Jafar Kiani\",\"doi\":\"10.18502/ijph.v53i4.15571\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><strong>Background: </strong>The lncRNAs has been linked to several malignancies, including breast cancer. Our objective was to investigate the impact of urothelial carcinoma associated 1 (<i>UCA1</i>) on cellular growth and death by a CRISPR/Cas9 knockdown technique.</p><p><strong>Methods: </strong>In 2020, the CHOPCHOP program was utilized to design two sgRNAs targeting the <i>UCA ge</i>ne. 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 <i>UCA1</i> and miR-143.</p><p><strong>Results: </strong>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 <i>Caspase-3</i> gene expression (<i>P</i><0.001). When WT-<i>UCA1</i> and miR-143 were co-transfected, the luciferase activity was drastically decreased.</p><p><strong>Conclusion: </strong>One very effective method of regulating cellular proliferation in vitro is the deletion of <i>UCA1</i>, which CRISPR/Cas9 accomplishes.</p>\",\"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}","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}
Long non-coding RNA UCA1 Knockdown Assisted by CRISPR/Cas9 in Female Cancer Cell Lines Increases Mir-143 Tumor-Suppressor.
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.
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.
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