Figure 1.
Identification of ZNF471 silenced by promoter methylation in ESCC cell lines. (A) A typical CpG island spanning ZNF471 (CpG Island Searcher). Each vertical bar represents a single CpG site. (B) ZNF471 expression and methylation status in ESCC cell lines. The RNA integrity of these samples was confirmed by GAPDH tests, as shown in our other publications 20. M, methylated; U, unmethylated. (C, D) ZNF471 expression and methylation status with 5-aza-2-deoxycytidine (Aza) and trichostatin A (TSA) treatments in ESCC cell lines. Demethylation was measured by real time quantitative MSP (qMSP). M, methylated; U, unmethylated. Dunnett's t-test was used. (E) ZNF471 expression in primary ESCC (n=16) and paired adjacent non-cancerous tissues (n=16) by qRT-PCR. Student's test was used. Data are presented as the mean ± SD. (F) ZNF471 methylation in primary ESCC tissues (n=147), adjacent non-cancerous tissues (n=89) and normal tissues (n=3), measured by MSP. M methylated, U unmethylated. Gel images showed were just representational graphs, not for all gel images. *p<0.05, **p<0.01, ***p<0.001.