scholarly journals Method for optimizing methylation-specific PCR

BioTechniques ◽  
2003 ◽  
Vol 35 (1) ◽  
pp. 32-33 ◽  
Author(s):  
Eric Smith ◽  
Tina Bianco-Miotto ◽  
Paul Drew ◽  
David Watson
Keyword(s):  
Methylation Specific Pcr ◽  
Specific Pcr

SUV39H1-Mediated DNMT1 is Participated in the Epigenetic Regulation of Smad3 in Cervical Cancer

2020 ◽  
Vol 20 ◽  
Author(s):  
Li Zhang ◽  
Sijuan Tian ◽  
Minyi Zhao ◽  
Ting Yang ◽  
Shimin Quan ◽  
...  
Keyword(s):  
Cervical Cancer ◽  
Cell Lines ◽  
Epigenetic Regulation ◽  
Promoter Region ◽  
Methylation Status ◽  
Regulation Mechanism ◽  
Methylation Specific Pcr ◽  
Specific Pcr ◽  
Immune Factor ◽  
Cancer Tissues

Background: Smad3 is a pivotal intracellular mediator for participating in the activation of multiple immune signal pathway. Objective: The epigenetic regulation mechanism of the positive immune factor Smad3 in cervical cancer remains unknown. Therefore, the epigenetic regulation on Smad3 is investigated in this study. Methods: The methylation status of SMAD3 was detected by Methylation-specific PCR (MS-PCR) and Quantitative Methylation-specific PCR (MS-qPCR) in cervical cancer tissues and cell lines. The underlying molecular mechanisms of SUV39H1-DNMT1-Smad3 regulation was elucidated using cervical cancer cell lines containing siRNA or/and overexpression system. Confirmation of the regulation of DNMT1 by SUV39H1 used Chromatin immunoprecipitation-qPCR (ChIP-qPCR). The statistical methods used for comparing samples between groups were paired t tests and one-way ANOVAs. Results: H3K9me3 protein which regulated by SUV39H1 directly interacts with the DNMT1 promoter region to regulate its expression in cervical cancer cells, resulting in the reduce expression of the downstream target gene DNMT1. In addition, DNMT1 mediates the epigenetic modulation of the SMAD3 gene by directly binding to its promoter region. The depletion of DNMT1 effectively restores the expression of Smad3 in vitro. Moreover, in an in vivo assay, the expression profile of SUV39H1-DNMT1 was found to correlate with Smad3 expression in accordance with the expression at the cellular level. Notably, the promoter region of SMAD3 was hypermethylated in cervical cancer tissues, and this hypermethylation inhibits the subsequent gene expression. Conclusion: These results indicate that SUV39H1-DNMT1 is a crucial Smad3 regulatory axis in cervical cancer. SUV39H1-DNMT1 axis may provide a potential therapeutic target for the treatment of cervical cancer.

scholarly journals Quantitative Multiplex Methylation-Specific PCR Analysis Doubles Detection of Tumor Cells in Breast Ductal Fluid

2006 ◽  
Vol 12 (11) ◽  
pp. 3306-3310 ◽  
Author(s):  
Mary Jo Fackler ◽  
Kara Malone ◽  
Zhe Zhang ◽  
Eric Schilling ◽  
Elizabeth Garrett-Mayer ◽  
...  
Keyword(s):  
Tumor Cells ◽  
Pcr Analysis ◽  
Methylation Specific Pcr ◽  
Specific Pcr ◽  
Ductal Fluid

Methylation-Specific PCR

2009 ◽  
pp. 305-323 ◽  
Author(s):  
Julien D. F. Licchesi ◽  
James G. Herman
Keyword(s):  
Methylation Specific Pcr ◽  
Specific Pcr

Prognosis of glioblastoma with faint MGMT methylation-specific PCR product

2015 ◽  
Vol 122 (1) ◽  
pp. 179-188 ◽  
Author(s):  
Chih-Yi Hsu ◽  
Hsiang-Ling Ho ◽  
Shih-Chieh Lin ◽  
Yi-Chun Chang-Chien ◽  
Ming-Hsiung Chen ◽  
...  
Keyword(s):  
Mgmt Methylation ◽  
Methylation Specific Pcr ◽  
Specific Pcr ◽  
Pcr Product
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