| Objective:Esophageal carcinoma is a kind of malignant tumor that emerges epithelial tissue of esophagus, and most kind of the esophageal carcinoma is esophageal squamous cell carcinoma (ESCC) in the developing country. The morbility and mortality of esophageal carcinoma is the highest in the world in Tai Hang mountain rang of Northern China. The disease incidence of hidden when its clinical symptoms are delayed There are a mount of people die of esophageal carcinoma every year. Study on pathogenesis becomes more important in order to decrease the morbility and mortality of esophageal carcinoma.Recently, the DNA methylation in pathogenesis of tumor has been the hot spot. The purpose of this study is to detect the methylation and the mRNA expression of wif-1, dkk3, sfrp1, sfrp2, sfrp4 and sfrp5 gene in ESCC and corresponding normal tissues, to approach the relationship between gene methylation and the carcinogenesis, diversion and the differentiation degree of ESCC, and to provide a new theory and experiment evidence for pathogenesy and clinical prognosis of esophageal squamous cell carcinoma.Methods:1.We detected the methylation status of the 5' CpG island of wif-1, dkk3, sfrp1, sfrp2, sfrp4 and sfrp5 gene in 65 cases of ESCC and 30 cases of corresponding normal tissues using methylation specific PCR (MSP) method.2.We examined the mRNA expression level of wif-1, dkk3, sfrp1, sfrp2, sfrp4 and sfrp5 gene in 20 cases of ESCC and 10 cases of corresponding normal tissues by reverse transcriptase PCR (RT-PCR) method.3.The data of experiment was analyzed by SPSS 13.0.Results:1.The methylation status of wif-1, dkk3, sfrp1, sfrp2, sfrp4 and sfrp5 gene in 65 cases of ESCC and 30 cases of corresponding normal tissues, and the relationship between their methylation and clinical pathology data were analyzed..wif-1gene was methylated in 18 of 65 (27.7%) tumor specimens, which was significantly higher than that in corresponding normal tissues 0% (0/30) (p<0.05). dkk3 gene was methylated in 21 of 65 (32.3%) tumor specimens, which was significantly higher than that in corresponding normal tissues 0% (0/30) (p<0.05). sfrp1 gene was methylated in 44 of 65 (67.7%) tumor specimens, which was significantly higher than that in corresponding normal tissues 43.3% (13/30) (p<0.05). sfrp2 gene was methylated in 45 of 65 (69.2%) tumor specimens, which was significantly higher than that in corresponding normal tissues 36.7% (11/30) (p<0.05). sfrp4 gene was methylated in 43 of 65 (66.2%) tumor specimens, which was significantly higher than that in corresponding normal tissues 6.7% (2/30) (p<0.05). sfrp5 gene was methylated in 37 of 65 (57.0%) tumor specimens, which was significantly higher than that in corresponding normal tissues 6.7% (2/30) (p<0.05).The methylation status of wif-1, dkk3, sfrp1, sfrp2 and sfrp5 gene have no relationship with clinical pathology data, such as, Gender, Age, Clinical stage, pathological differentiation, Lymph node status.Methylation frequence of sfrp4 gene in poor differentiation group 30.0% was significantly lower than moderate and poor-moderate differentiation groups 74.5% (p=0.006, p<0.05).2.The mRNA expression of wif-1, dkk3, sfrp1, sfrp2, sfrp4 and sfrp5 gene in 20 cases of ESCC and 10 cases of corresponding normal tissues.wif-1gene mRNA expression in ESCC group (0.67±0.32) was significantly lower than that in corresponding normal group (0.95±0.23) (p<0.05). dkk3 gene mRNA expression in ESCC group (0.62±0.22) was significantly lower than that in corresponding normal group (0.86±0.33) (p<0.05). sfrp1 gene mRNA expression in ESCC group (0.66±0.35) was significantly lower than corresponding normal group (0.93±0.19) (p<0.05). sfrp2 gene mRNA expression in ESCC group (0.62±0.19) was significantly lower than that in corresponding normal group (0.89±0.32) (p<0.05). sfrp4 gene mRNA expression in ESCC group (0.67±0.29) was significantly lower than that in corresponding normal group (0.93±0.16) (p<0.05). sfrp5 gene mRNA expression in ESCC group (0.59±0.23) was significantly lower than that in corresponding normal group (0.92±0.36) (p<0.05).3.The relationship between methylation of wif-1, dkk3, sfrp1, sfrp2, sfrp4 and sfrp5 gene and mRNA expression in 20 cases of esophageal squamous cell carcinoma tissues.wif-1, dkk3, sfrp1, sfrp2, sfrp4 and sfrp5 gene mRNA expression are lower in methylated ESCC tissues than that in unmethylated ESCC tissues, but the differences have no significance (p>0.05).Conclusions:1.The methylation of wif-1, dkk3, sfrp1, sfrp2, sfrp4 and sfrp5 gene in ESCC were all significantly higher than that in corresponding normal tissues. The results indicated that the methylation of wif-1, dkk3, sfrp1, sfrp2, sfrp4 and sfrp5 gene may be related with oncogenesis of ESCC.2.The methylation frequence of sfrp4 in poor differentiation group was significantly decrease than that in moderate and poor-moderate differentiation groups. The results indicated that sfrp4 may be related with the malignant biological behavior of ESCC.3.The mRNA expression of wif-1, dkk3, sfrp1, sfrp2, sfrp4 and sfrp5 gene in ESCC were all significantly decreased than that in corresponding normal tissues. The results indicated that abnormal expression of the six genes mRNA may be related with oncogenesis of ESCC.4.The expression difference of wif-1, dkk3, sfrp1, sfrp2, sfrp4 and sfrp5 gene mRNA was not significantly between methylated ESCC tissues and unmethylated ESCC tissues, which indicated that methylation maybe not the main reason that cause the abnormal expression of mRNA.
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