New primers for methylation-specific polymerase chain reaction enhance specificity of detecting STAT1 methylation
Objective: Signal transducer and activator of transcription (STAT)1 is a key tumor suppressor, which is always methylated in a variety of human cancers. However, nonspecific primers for the detection of specific promoter hypermethylation of STAT1 gene can lead to false-positive or false-negative res...
| Main Authors: | , , , , , , |
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| Format: | Article |
| Language: | English |
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Elsevier
2012-03-01
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| Series: | Taiwanese Journal of Obstetrics & Gynecology |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S1028455912000101 |
| _version_ | 1818912228655497216 |
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| author | Ming-Cheng Chang Ying-Cheng Chiang Chih-Ming Ho Yu-Li Chen Chi-An Chen Wen-Fang Cheng Cheng-Yang Chou |
| author_facet | Ming-Cheng Chang Ying-Cheng Chiang Chih-Ming Ho Yu-Li Chen Chi-An Chen Wen-Fang Cheng Cheng-Yang Chou |
| author_sort | Ming-Cheng Chang |
| collection | DOAJ |
| description | Objective: Signal transducer and activator of transcription (STAT)1 is a key tumor suppressor, which is always methylated in a variety of human cancers. However, nonspecific primers for the detection of specific promoter hypermethylation of STAT1 gene can lead to false-positive or false-negative results for gene methylation.
Materials and Methods: We designed new primers for the detection of STAT1 methylation and compared the sensitivities and specificities of these new primers with prior published primers by methylation-specific polymerase chain reaction (PCR) from ovarian clear cell carcinomas. The mRNA expression levels of STAT1 in these cancerous tissues were also evaluated by reverse-transcriptase PCR and correlated with the results of promoter methylation of STAT1 gene.
Results: Nine (39%) of the 23 samples detected by the new primers and 13 samples (56%) detected by prior published primers showed STAT1 methylation. A direct DNA sequencing test revealed that four of the 13 samples (30.8%) showed false positivity for STAT1 methylation using the prior published primers. In contrast, none of the nine samples was false-positive for the detection of STAT1 methylation using the new primers. The new primers for the detection of STAT1 methylation showed 100% specificity and 100% sensitivity without false positivity.
Conclusion: Specific primers for methylation-specific PCR are mandatory for the accurate detection of STAT1 gene methylation. Besides, specific primers can generate correct interpretation of STAT1 gene methylation, and its correlation with the clinicopathological characteristics and outcome of cancer patients. |
| first_indexed | 2024-12-19T23:11:16Z |
| format | Article |
| id | doaj.art-071f3a1a72c64c80a0660c161a95962e |
| institution | Directory Open Access Journal |
| issn | 1028-4559 |
| language | English |
| last_indexed | 2024-12-19T23:11:16Z |
| publishDate | 2012-03-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Taiwanese Journal of Obstetrics & Gynecology |
| spelling | doaj.art-071f3a1a72c64c80a0660c161a95962e2022-12-21T20:02:13ZengElsevierTaiwanese Journal of Obstetrics & Gynecology1028-45592012-03-01511434910.1016/j.tjog.2012.01.009New primers for methylation-specific polymerase chain reaction enhance specificity of detecting STAT1 methylationMing-Cheng Chang0Ying-Cheng Chiang1Chih-Ming Ho2Yu-Li Chen3Chi-An Chen4Wen-Fang Cheng5Cheng-Yang Chou6Department of Obstetrics and Gynecology, Institute of Clinical Medicine, College of Medicine, National Taiwan University, Taipei, TaiwanDepartment of Obstetrics and Gynecology, National Taiwan University Hospital Yun-Lin Branch, Douliou City, Yunlin County, TaiwanGynecologic Cancer Center, Department of Obstetrics and Gynecology, Cathay General Hospital, Taipei, TaiwanGynecologic Cancer Center, Department of Obstetrics and Gynecology, Cathay General Hospital, Taipei, TaiwanDepartment of Obstetrics and Gynecology, Institute of Clinical Medicine, College of Medicine, National Taiwan University, Taipei, TaiwanDepartment of Obstetrics and Gynecology, Institute of Clinical Medicine, College of Medicine, National Taiwan University, Taipei, TaiwanDepartment of Obstetrics and Gynecology, College of Medicine, National Cheng Kung University, Tainan, TaiwanObjective: Signal transducer and activator of transcription (STAT)1 is a key tumor suppressor, which is always methylated in a variety of human cancers. However, nonspecific primers for the detection of specific promoter hypermethylation of STAT1 gene can lead to false-positive or false-negative results for gene methylation. Materials and Methods: We designed new primers for the detection of STAT1 methylation and compared the sensitivities and specificities of these new primers with prior published primers by methylation-specific polymerase chain reaction (PCR) from ovarian clear cell carcinomas. The mRNA expression levels of STAT1 in these cancerous tissues were also evaluated by reverse-transcriptase PCR and correlated with the results of promoter methylation of STAT1 gene. Results: Nine (39%) of the 23 samples detected by the new primers and 13 samples (56%) detected by prior published primers showed STAT1 methylation. A direct DNA sequencing test revealed that four of the 13 samples (30.8%) showed false positivity for STAT1 methylation using the prior published primers. In contrast, none of the nine samples was false-positive for the detection of STAT1 methylation using the new primers. The new primers for the detection of STAT1 methylation showed 100% specificity and 100% sensitivity without false positivity. Conclusion: Specific primers for methylation-specific PCR are mandatory for the accurate detection of STAT1 gene methylation. Besides, specific primers can generate correct interpretation of STAT1 gene methylation, and its correlation with the clinicopathological characteristics and outcome of cancer patients.http://www.sciencedirect.com/science/article/pii/S1028455912000101methylationpolymerase chain reactionsignal transducer and activator of transcription 1 |
| spellingShingle | Ming-Cheng Chang Ying-Cheng Chiang Chih-Ming Ho Yu-Li Chen Chi-An Chen Wen-Fang Cheng Cheng-Yang Chou New primers for methylation-specific polymerase chain reaction enhance specificity of detecting STAT1 methylation Taiwanese Journal of Obstetrics & Gynecology methylation polymerase chain reaction signal transducer and activator of transcription 1 |
| title | New primers for methylation-specific polymerase chain reaction enhance specificity of detecting STAT1 methylation |
| title_full | New primers for methylation-specific polymerase chain reaction enhance specificity of detecting STAT1 methylation |
| title_fullStr | New primers for methylation-specific polymerase chain reaction enhance specificity of detecting STAT1 methylation |
| title_full_unstemmed | New primers for methylation-specific polymerase chain reaction enhance specificity of detecting STAT1 methylation |
| title_short | New primers for methylation-specific polymerase chain reaction enhance specificity of detecting STAT1 methylation |
| title_sort | new primers for methylation specific polymerase chain reaction enhance specificity of detecting stat1 methylation |
| topic | methylation polymerase chain reaction signal transducer and activator of transcription 1 |
| url | http://www.sciencedirect.com/science/article/pii/S1028455912000101 |
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