{"title":"LBC固定类型和固定时间对胰腺癌细胞分子分析的影响:细胞形态、抗原性和核酸的比较研究。","authors":"Junya Izuhara, Kazuki Kanayama","doi":"10.4103/joc.joc_13_22","DOIUrl":null,"url":null,"abstract":"<p><strong>Introduction: </strong>Liquid-based cytology (LBC) has been widely used since 2000. Next-Generation Sequencing (NGS) analysis of residual specimens in LBC fixative may also be performed for pancreatic cancer in the near future. We examined cell morphology, antigenicity and nucleic acids in pancreatic cancer cells at different fixation times using two types of LBC fixatives.</p><p><strong>Methods: </strong>PANC-1 cells were fixed in 1 ml CytoRich Red (CR), CytoRich Blue (CB), 95% ethanol (95% AL) or 10% neutral buffered formalin (10% NBF) and evaluated for cell area, antigenicity and nucleic acids with fixation times of 1 hour and 1, 3, 9, and 14 days. Antigenicity was evaluated by immunocytochemical staining for p53 and CK20, and nucleic acid fragmentation was assessed by real-time PCR.</p><p><strong>Results: </strong>There was no difference in total cell area between 1 hour and 14 day fixation times for the CR group, but the CB group showed cell contraction with 9 days fixation. In immunocytochemical staining, the CR group showed high p53 and CK20 positivity even after 14 days fixation. The CB group had a lower p53 positive rate than the CR group from 1 hour fixation. For nucleic acid fragmentation, Ct values for the CR group increased with fixation time. The CB group had consistently low Ct values.</p><p><strong>Conclusion: </strong>Different LBC fixatives and fixation time can have varying effects on cell morphology, antigenicity and nucleic acids in pancreatic cancer cells. Therefore, fixative type and fixation time should be considered for molecular testing on residual samples in LBC fixatives.</p>","PeriodicalId":50217,"journal":{"name":"Journal of Cytology","volume":"39 2","pages":"66-71"},"PeriodicalIF":1.0000,"publicationDate":"2022-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9261998/pdf/","citationCount":"0","resultStr":"{\"title\":\"Impact of LBC Fixative Type and Fixation Time on Molecular Analysis of Pancreatic Cancer Cells: A Comparative Study of Cell Morphology, Antigenicity and Nucleic Acids.\",\"authors\":\"Junya Izuhara, Kazuki Kanayama\",\"doi\":\"10.4103/joc.joc_13_22\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><strong>Introduction: </strong>Liquid-based cytology (LBC) has been widely used since 2000. Next-Generation Sequencing (NGS) analysis of residual specimens in LBC fixative may also be performed for pancreatic cancer in the near future. We examined cell morphology, antigenicity and nucleic acids in pancreatic cancer cells at different fixation times using two types of LBC fixatives.</p><p><strong>Methods: </strong>PANC-1 cells were fixed in 1 ml CytoRich Red (CR), CytoRich Blue (CB), 95% ethanol (95% AL) or 10% neutral buffered formalin (10% NBF) and evaluated for cell area, antigenicity and nucleic acids with fixation times of 1 hour and 1, 3, 9, and 14 days. Antigenicity was evaluated by immunocytochemical staining for p53 and CK20, and nucleic acid fragmentation was assessed by real-time PCR.</p><p><strong>Results: </strong>There was no difference in total cell area between 1 hour and 14 day fixation times for the CR group, but the CB group showed cell contraction with 9 days fixation. In immunocytochemical staining, the CR group showed high p53 and CK20 positivity even after 14 days fixation. The CB group had a lower p53 positive rate than the CR group from 1 hour fixation. For nucleic acid fragmentation, Ct values for the CR group increased with fixation time. The CB group had consistently low Ct values.</p><p><strong>Conclusion: </strong>Different LBC fixatives and fixation time can have varying effects on cell morphology, antigenicity and nucleic acids in pancreatic cancer cells. Therefore, fixative type and fixation time should be considered for molecular testing on residual samples in LBC fixatives.</p>\",\"PeriodicalId\":50217,\"journal\":{\"name\":\"Journal of Cytology\",\"volume\":\"39 2\",\"pages\":\"66-71\"},\"PeriodicalIF\":1.0000,\"publicationDate\":\"2022-04-01\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9261998/pdf/\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Journal of Cytology\",\"FirstCategoryId\":\"3\",\"ListUrlMain\":\"https://doi.org/10.4103/joc.joc_13_22\",\"RegionNum\":4,\"RegionCategory\":\"医学\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"2022/5/20 0:00:00\",\"PubModel\":\"Epub\",\"JCR\":\"Q4\",\"JCRName\":\"MEDICAL LABORATORY TECHNOLOGY\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Journal of Cytology","FirstCategoryId":"3","ListUrlMain":"https://doi.org/10.4103/joc.joc_13_22","RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"2022/5/20 0:00:00","PubModel":"Epub","JCR":"Q4","JCRName":"MEDICAL LABORATORY TECHNOLOGY","Score":null,"Total":0}
引用次数: 0
摘要
自2000年以来,液体细胞学(LBC)得到了广泛的应用。在不久的将来,LBC固定液中残留标本的下一代测序(NGS)分析也可能用于胰腺癌。我们使用两种类型的LBC固定剂在不同的固定时间检测胰腺癌细胞的细胞形态、抗原性和核酸。方法:将PANC-1细胞固定于1ml CytoRich Red (CR)、CytoRich Blue (CB)、95%乙醇(95% AL)或10%中性缓冲福尔马林(10% NBF)中,固定时间分别为1小时、1、3、9、14天,评估细胞面积、抗原性和核酸。免疫细胞化学染色检测p53和CK20的抗原性,实时荧光定量PCR检测核酸片段性。结果:CR组细胞总面积在固定时间1 h和14 d无差异,而CB组在固定时间9 d出现细胞收缩。在免疫细胞化学染色中,CR组在固定14天后仍显示高p53和CK20阳性。固定1 h后,CB组p53阳性率低于CR组。CR组的Ct值随固定时间的延长而增加。CB组的Ct值一直很低。结论:不同的LBC固定剂和固定时间对胰腺癌细胞形态、抗原性和核酸有不同的影响。因此,对LBC固定液中残留样品进行分子检测时应考虑固定液类型和固定时间。
Impact of LBC Fixative Type and Fixation Time on Molecular Analysis of Pancreatic Cancer Cells: A Comparative Study of Cell Morphology, Antigenicity and Nucleic Acids.
Introduction: Liquid-based cytology (LBC) has been widely used since 2000. Next-Generation Sequencing (NGS) analysis of residual specimens in LBC fixative may also be performed for pancreatic cancer in the near future. We examined cell morphology, antigenicity and nucleic acids in pancreatic cancer cells at different fixation times using two types of LBC fixatives.
Methods: PANC-1 cells were fixed in 1 ml CytoRich Red (CR), CytoRich Blue (CB), 95% ethanol (95% AL) or 10% neutral buffered formalin (10% NBF) and evaluated for cell area, antigenicity and nucleic acids with fixation times of 1 hour and 1, 3, 9, and 14 days. Antigenicity was evaluated by immunocytochemical staining for p53 and CK20, and nucleic acid fragmentation was assessed by real-time PCR.
Results: There was no difference in total cell area between 1 hour and 14 day fixation times for the CR group, but the CB group showed cell contraction with 9 days fixation. In immunocytochemical staining, the CR group showed high p53 and CK20 positivity even after 14 days fixation. The CB group had a lower p53 positive rate than the CR group from 1 hour fixation. For nucleic acid fragmentation, Ct values for the CR group increased with fixation time. The CB group had consistently low Ct values.
Conclusion: Different LBC fixatives and fixation time can have varying effects on cell morphology, antigenicity and nucleic acids in pancreatic cancer cells. Therefore, fixative type and fixation time should be considered for molecular testing on residual samples in LBC fixatives.
期刊介绍:
The Journal of Cytology is the official Quarterly publication of the Indian Academy of Cytologists. It is in the 25th year of publication in the year 2008. The journal covers all aspects of diagnostic cytology, including fine needle aspiration cytology, gynecological and non-gynecological cytology. Articles on ancillary techniques, like cytochemistry, immunocytochemistry, electron microscopy, molecular cytopathology, as applied to cytological material are also welcome. The journal gives preference to clinically oriented studies over experimental and animal studies. The Journal would publish peer-reviewed original research papers, case reports, systematic reviews, meta-analysis, and debates.