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Sample Preparation to Bioinformatics Analysis of DNA Methylation: Association Strategy for Obesity and Related Trait Studies
Published on: May 6, 2022
Association between CpG island methylation of the WWOX gene and its expression in breast cancers
1Department of Breast Surgery, Jinan Central Hospital, Medical College, Jinan, China.
Objectives:
The aim of the present study was to evaluate the expression of the WWOX gene in breast carcinoma and to analyze the relationship between its expression and methylation.
Methods:
Methylation of the WWOX promoter and exon 1 as well as WWOX expression were determined by methylation-specific PCR, reverse transcription PCR and immunohistochemistry in 20 breast cancer tissue samples and corresponding nonneoplastic tissues. The expression of WWOX protein was evaluated by immunohistochemistry in 2 breast cancer cell lines.
Results:
WWOX mRNA expression was reduced in breast cancer tissues as compared to adjacent tissues (p < 0.001). WWOX mRNA and protein expression was significantly reduced in breast cancer tissues with methylated WWOX CpG islands when compared to tissues without methylation (p < 0.001 and p = 0.008, respectively). Immunohistochemical analysis also showed that WWOX levels were significantly decreased in tissues with WWOX methylation (p = 0.001). WWOX mRNA and protein expression was greatly reduced in MDA-MB-231 cells, partly due to the methylation of WWOX CpG islands, and recovered after hydralazine treatment.
Conclusions:
Differential regulation of the WWOX gene plays an important role in the carcinogenesis of breast cancer, due partially to methylation of the WWOX DNA CpG islands.
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