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 journalArticlepeer-review

3 Citations (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

All Science Journal Classification (ASJC) codes

  • Obstetrics and Gynaecology

Fingerprint Dive into the research topics of 'New primers for methylation-specific polymerase chain reaction enhance specificity of detecting STAT1 methylation'. Together they form a unique fingerprint.

Cite this