A. RT-PCR showed that MUCs up-regulated the expression of epithelial genes Cdh1, Krt8, and Krt18 following 5-aza-CdR treatment, and the relative expression levels of studied genes were related to 5-aza-CdR doses.
B. Quantification PCR showed normalized relative expressions of Cdh1 and Krt18 of 4 μM 5-aza-CdR-treated MUCs were 13.64 ± 0.05 and 13.57 ± 3.68 fold higher than those of control MUCs respectively, which is statistically significant (P<0.05; Student’s t-test).
C. Nested-Methylation Specific PCR (Nested-MSP) showed that the normalized expression of methylated Cdh1 in control MUCs was higher than unmethylated Cdh1. After 4 μM 5-aza-CdR treatment, the normalized expression of methylated Cdh1 was significantly down-regulated and lower than the expression of unmethylated Cdh1. The normalized relative expression of methylated Cdh1 was remarkably decreased following 5-aza-CdR treatment.