浙江医学2018，Vol.40Issue(4)：333-338,6.DOI:10.12056/j.issn.1006-2785.2017.40.4.2017-2955
Wnt6、Wnt3A、LEF1和FZD1基因启动子甲基化在结直肠癌筛查及早期诊断中的临床应用
The clinical application of detecting Wnt6, Wnt3A, LEF1 and FZD1 gene methylation in the screening, early diagnosis of colorectal cancer
摘要
Abstract
Objective To explore clinical application of Wnt6,Wnt3A,LEF1 and FZD1 gene methylation in colorectal cancer (CRC) screening and early diagnosis.Methods Fifty-five patients with colorectal cancer and 55 healthy people were enrolled in the Affiliated Hospital of Ningbo University Medical College during December 2015 to December 2016.The DNA of colorectal cancer tissue,blood and stool were isolated by QIAGEN kit,and the DNA of the three control groups were also isolated by QIAGEN kit.MS-PCR (methylation-specific PCR) was applied to analyze the promoter hypermethylation of Wnt6,Wnt3A,LEF1 and FZD1 genes.The mRNA levels of Wnt6,Wnt3A,LEF1 and FZD1 were detected by RT-PCR and the protein expression levels of Wnt6,Wnt3A,LEF1 and FZD1 were detected by Western-blot,to explore Whether the promoter hypermethylation of theWnt6,Wnt3A,LEF1 and FZD1 genes causes downregulation gene expression.Results A total of 110 cases of tissue,blood,and fecal abscission cells (DNA) were extracted to verify the existence of human genome DNA and complete MSP in 110 cases.The detection rates of Wnt6,Wnt3A,LEF1 and FZD1 promoter methylation in the DNA of fecal exfoliated cells were 69.1％,63.6％,54.5％ and 56.4％ respectively in the observation group.The detection rate of 4 gene promoter methylation of the observation group was 90.9％.The detection rates of Wnt6,Wnt3A,LEF1 and FZD1 promoter methylation in the DNA of fecal exfoliated cells were 0.0％,1.8％,0.0％ and 1.8％ espectively in the control group.The detection rate of 4 gene promoter methylation of the control group was 3.6％.he detection rates of Wnt6,Wnt3A,LEF1 and FZD1 promoter methylation in the DNA of fecal exfoliated cells were 80.9％,71.4％,57.1％ and 57.1％ for 21 cases of the stage Ⅰ-Ⅱ CRC respectively.The detection rate of 4 gene promoter methylation for stage Ⅰ to Ⅱ CRC was 90.5％.Methylation of Wnt6,Wnt3A,LEF1 and FZD1 genes in DNA of fecal exfoliated cells from CRC patients was not correlated with gender,age,distant metastasis,tumor location,TNM stage and lymph node metastasis (P ＞0.05).The methylation positive rates of Wnt6,Wnt3A,LEF1 and FZD1 genes in the colorectal tissues,blood samples and feces samples of the observation group and the control group were statistically significant (all P＜0.01).The mRNA and protein levels of Wnt6,Wnt3A,LEF1 and FZD1 gene promoter in the two groups were all down regulated,and the difference between the observation group and the control group was significant.Conclusion Detection of methylation of Wnt6,Wnt3A,LEF1 and FZD1 genes in DNA and exfoliated cells from blood and feces by MSP is a effective clinical method for early CRC screening.关键词
结直肠癌/甲基化特异性PCR/甲基化Key words
Colorectal cancer/MSP/Promoter methylation引用本文复制引用
杨齐华,叶国良,徐拔群,洪晓波..Wnt6、Wnt3A、LEF1和FZD1基因启动子甲基化在结直肠癌筛查及早期诊断中的临床应用[J].浙江医学,2018,40(4):333-338,6.基金项目
宁波市自然科学基金项目(2016A610123) （2016A610123）
