New primers for methylation-specific polymerase chain reaction enhance specificity of detecting STAT1 methylation

Ming Cheng Chang, Ying Cheng Chiang, Chih Ming Ho, Yu Li Chen, Chi An Chen, Wen Fang Cheng, Cheng-Yang Chou

Research output: Contribution to journalArticle

1 Citation (Scopus)

Abstract

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.

Original languageEnglish
Pages (from-to)43-49
Number of pages7
JournalTaiwanese Journal of Obstetrics and Gynecology
Volume51
Issue number1
DOIs
Publication statusPublished - 2012 Mar 1

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Methylation
Polymerase Chain Reaction
Genes
STAT1 Transcription Factor
Sensitivity and Specificity
Neoplasms
Reverse Transcriptase Polymerase Chain Reaction
DNA Sequence Analysis
Carcinoma
Messenger RNA

All Science Journal Classification (ASJC) codes

  • Obstetrics and Gynaecology

Cite this

Chang, Ming Cheng ; Chiang, Ying Cheng ; Ho, Chih Ming ; Chen, Yu Li ; Chen, Chi An ; Cheng, Wen Fang ; Chou, Cheng-Yang. / New primers for methylation-specific polymerase chain reaction enhance specificity of detecting STAT1 methylation. In: Taiwanese Journal of Obstetrics and Gynecology. 2012 ; Vol. 51, No. 1. pp. 43-49.
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abstract = "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.",
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New primers for methylation-specific polymerase chain reaction enhance specificity of detecting STAT1 methylation. / Chang, Ming Cheng; Chiang, Ying Cheng; Ho, Chih Ming; Chen, Yu Li; Chen, Chi An; Cheng, Wen Fang; Chou, Cheng-Yang.

In: Taiwanese Journal of Obstetrics and Gynecology, Vol. 51, No. 1, 01.03.2012, p. 43-49.

Research output: Contribution to journalArticle

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AU - Chiang, Ying Cheng

AU - Ho, Chih Ming

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AU - Chou, Cheng-Yang

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N2 - 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.

AB - 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.

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