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Semi‐quantitative Analysis of DNA Topoisomerase‐I mRNA Level Using Reverse Transcription‐Polymerase Chain Reaction in Cancer Cell Lines: Its Relation to Cytotoxicity against Camptothecin Derivative

Expression of DNA topoisomerase (Topo)‐I mRNA in various cancer cell lines was detected using the reverse transcription‐polymerase chain reaction (RT‐PCR) method. The cytoplasmic polyadenylated RNA isolated from cancer cell lines was reverse‐transcribed and the complementary DNA was amplified by PCR...

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Autores principales: Niwa, Kenji, Misao, Ryou, Hanabayashi, Takabiro, Morishita, Shigeo, Murase, Toshiko, Itoh, Miho, Itoh, Naoki, Mori, Hidehiro, Tamaya, Teruhiko
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Blackwell Publishing Ltd 1994
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5919567/
https://www.ncbi.nlm.nih.gov/pubmed/7523352
http://dx.doi.org/10.1111/j.1349-7006.1994.tb02960.x
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author Niwa, Kenji
Misao, Ryou
Hanabayashi, Takabiro
Morishita, Shigeo
Murase, Toshiko
Itoh, Miho
Itoh, Naoki
Mori, Hidehiro
Tamaya, Teruhiko
author_facet Niwa, Kenji
Misao, Ryou
Hanabayashi, Takabiro
Morishita, Shigeo
Murase, Toshiko
Itoh, Miho
Itoh, Naoki
Mori, Hidehiro
Tamaya, Teruhiko
author_sort Niwa, Kenji
collection PubMed
description Expression of DNA topoisomerase (Topo)‐I mRNA in various cancer cell lines was detected using the reverse transcription‐polymerase chain reaction (RT‐PCR) method. The cytoplasmic polyadenylated RNA isolated from cancer cell lines was reverse‐transcribed and the complementary DNA was amplified by PCR primed with Topo‐I specific primers. Fidelity of the amplified sequence was confirmed by restriction endonuclease digestion and Southern blot hybridization. The level of Topo‐I mRNA was correlated positively with the cytotoxicity of a Topo‐I inhibitor, a camptothecin derivative. This RT‐PCR method may be applicable to the assessment of sensitivity of cells to Topo‐I targeted drugs, especially when only small quantities of cell samples are available.
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spelling pubmed-59195672018-05-11 Semi‐quantitative Analysis of DNA Topoisomerase‐I mRNA Level Using Reverse Transcription‐Polymerase Chain Reaction in Cancer Cell Lines: Its Relation to Cytotoxicity against Camptothecin Derivative Niwa, Kenji Misao, Ryou Hanabayashi, Takabiro Morishita, Shigeo Murase, Toshiko Itoh, Miho Itoh, Naoki Mori, Hidehiro Tamaya, Teruhiko Jpn J Cancer Res Article Expression of DNA topoisomerase (Topo)‐I mRNA in various cancer cell lines was detected using the reverse transcription‐polymerase chain reaction (RT‐PCR) method. The cytoplasmic polyadenylated RNA isolated from cancer cell lines was reverse‐transcribed and the complementary DNA was amplified by PCR primed with Topo‐I specific primers. Fidelity of the amplified sequence was confirmed by restriction endonuclease digestion and Southern blot hybridization. The level of Topo‐I mRNA was correlated positively with the cytotoxicity of a Topo‐I inhibitor, a camptothecin derivative. This RT‐PCR method may be applicable to the assessment of sensitivity of cells to Topo‐I targeted drugs, especially when only small quantities of cell samples are available. Blackwell Publishing Ltd 1994-08 /pmc/articles/PMC5919567/ /pubmed/7523352 http://dx.doi.org/10.1111/j.1349-7006.1994.tb02960.x Text en
spellingShingle Article
Niwa, Kenji
Misao, Ryou
Hanabayashi, Takabiro
Morishita, Shigeo
Murase, Toshiko
Itoh, Miho
Itoh, Naoki
Mori, Hidehiro
Tamaya, Teruhiko
Semi‐quantitative Analysis of DNA Topoisomerase‐I mRNA Level Using Reverse Transcription‐Polymerase Chain Reaction in Cancer Cell Lines: Its Relation to Cytotoxicity against Camptothecin Derivative
title Semi‐quantitative Analysis of DNA Topoisomerase‐I mRNA Level Using Reverse Transcription‐Polymerase Chain Reaction in Cancer Cell Lines: Its Relation to Cytotoxicity against Camptothecin Derivative
title_full Semi‐quantitative Analysis of DNA Topoisomerase‐I mRNA Level Using Reverse Transcription‐Polymerase Chain Reaction in Cancer Cell Lines: Its Relation to Cytotoxicity against Camptothecin Derivative
title_fullStr Semi‐quantitative Analysis of DNA Topoisomerase‐I mRNA Level Using Reverse Transcription‐Polymerase Chain Reaction in Cancer Cell Lines: Its Relation to Cytotoxicity against Camptothecin Derivative
title_full_unstemmed Semi‐quantitative Analysis of DNA Topoisomerase‐I mRNA Level Using Reverse Transcription‐Polymerase Chain Reaction in Cancer Cell Lines: Its Relation to Cytotoxicity against Camptothecin Derivative
title_short Semi‐quantitative Analysis of DNA Topoisomerase‐I mRNA Level Using Reverse Transcription‐Polymerase Chain Reaction in Cancer Cell Lines: Its Relation to Cytotoxicity against Camptothecin Derivative
title_sort semi‐quantitative analysis of dna topoisomerase‐i mrna level using reverse transcription‐polymerase chain reaction in cancer cell lines: its relation to cytotoxicity against camptothecin derivative
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5919567/
https://www.ncbi.nlm.nih.gov/pubmed/7523352
http://dx.doi.org/10.1111/j.1349-7006.1994.tb02960.x
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