{"title":"CRISPR/Cas9-mediated high-mobility group A2 knockout inhibits cell proliferation and invasion in papillary thyroid carcinoma cells","authors":"Shan Jin, Hong Yong, Yousheng Liu, Wuyuntu Bao","doi":"10.1016/j.advms.2023.10.001","DOIUrl":null,"url":null,"abstract":"<div><h3>Purpose</h3><p>Metastasis<span><span> and recurrence are the prognostic risk factor in patients<span> with thyroid carcinoma. High-mobility group A2 (HMGA2) protein plays a crucial role in </span></span>papillary thyroid carcinoma<span> (PTC) metastasis. The aim of this study was to investigate the mechanisms underlying the HMGA2 effect on PTC cell proliferation and invasion.</span></span></p></div><div><h3>Materials and methods</h3><p>We used the CRISPR/Cas9 system to perform knockout of the <em>HMGA2</em><span> gene in the human PTC cell line TPC-1. The knockout monoclonal cells were screened and verified by PCR analysis and genomic sequencing. Cell proliferation was examined after the knockout of the </span><em>HMGA2</em><span> gene using cell counting kit-8 (CCK-8) assays. Furthermore, cell migration and invasion after the knockout were examined by cell scratch tests. Additionally, the changes in cell cycle and apoptosis after the knockout were detected by flow cytometry.</span></p></div><div><h3>Results</h3><p>The results of the PCR analysis and the genomic sequencing confirmed that the human PTC TPC-1 cell line with knockout of <em>HMGA2</em> gene was successfully established. The knockout of the <em>HMGA2</em> gene significantly reduced the cell proliferation, growth, and invasion. Meanwhile, the knockout of the <em>HMGA2</em> gene delayed the conversion of the G2/M phase and promoted cell necrosis.</p></div><div><h3>Conclusion</h3><p>The CRISPR/Cas9-mediated <em>HMGA2</em> knockout in the TPC-1 cell line inhibited cell proliferation and invasion, which might be due to the blockage of the cell cycle in the G2/M phase and the promotion of cell necrosis.</p></div>","PeriodicalId":7347,"journal":{"name":"Advances in medical sciences","volume":"68 2","pages":"Pages 409-416"},"PeriodicalIF":2.5000,"publicationDate":"2023-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Advances in medical sciences","FirstCategoryId":"3","ListUrlMain":"https://www.sciencedirect.com/science/article/pii/S189611262300041X","RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q3","JCRName":"MEDICINE, RESEARCH & EXPERIMENTAL","Score":null,"Total":0}
引用次数: 0
Abstract
Purpose
Metastasis and recurrence are the prognostic risk factor in patients with thyroid carcinoma. High-mobility group A2 (HMGA2) protein plays a crucial role in papillary thyroid carcinoma (PTC) metastasis. The aim of this study was to investigate the mechanisms underlying the HMGA2 effect on PTC cell proliferation and invasion.
Materials and methods
We used the CRISPR/Cas9 system to perform knockout of the HMGA2 gene in the human PTC cell line TPC-1. The knockout monoclonal cells were screened and verified by PCR analysis and genomic sequencing. Cell proliferation was examined after the knockout of the HMGA2 gene using cell counting kit-8 (CCK-8) assays. Furthermore, cell migration and invasion after the knockout were examined by cell scratch tests. Additionally, the changes in cell cycle and apoptosis after the knockout were detected by flow cytometry.
Results
The results of the PCR analysis and the genomic sequencing confirmed that the human PTC TPC-1 cell line with knockout of HMGA2 gene was successfully established. The knockout of the HMGA2 gene significantly reduced the cell proliferation, growth, and invasion. Meanwhile, the knockout of the HMGA2 gene delayed the conversion of the G2/M phase and promoted cell necrosis.
Conclusion
The CRISPR/Cas9-mediated HMGA2 knockout in the TPC-1 cell line inhibited cell proliferation and invasion, which might be due to the blockage of the cell cycle in the G2/M phase and the promotion of cell necrosis.
期刊介绍：
Advances in Medical Sciences is an international, peer-reviewed journal that welcomes original research articles and reviews on current advances in life sciences, preclinical and clinical medicine, and related disciplines.
The Journal’s primary aim is to make every effort to contribute to progress in medical sciences. The strive is to bridge laboratory and clinical settings with cutting edge research findings and new developments.
Advances in Medical Sciences publishes articles which bring novel insights into diagnostic and molecular imaging, offering essential prior knowledge for diagnosis and treatment indispensable in all areas of medical sciences. It also publishes articles on pathological sciences giving foundation knowledge on the overall study of human diseases. Through its publications Advances in Medical Sciences also stresses the importance of pharmaceutical sciences as a rapidly and ever expanding area of research on drug design, development, action and evaluation contributing significantly to a variety of scientific disciplines.
The journal welcomes submissions from the following disciplines:
General and internal medicine,
Cancer research,
Genetics,
Endocrinology,
Gastroenterology,
Cardiology and Cardiovascular Medicine,
Immunology and Allergy,
Pathology and Forensic Medicine,
Cell and molecular Biology,
Haematology,
Biochemistry,
Clinical and Experimental Pathology.
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