Clinical value of methylation of plasma adenomatous polyposis coli gene in the molecular diagnosis of hepatocellular carcinoma / 肿瘤

ABSTRACT

Objective: To establish a method of methylation-sensitive restriction enzymes-quantitative PCR (MSRE-qPCR) for methylation analysis of adenomatous polyposis coli (APC ) gene, and to further assess the clinical value of plasma methylation detection by using this method in diagnosis of hepatocellular carcinoma (HCC). Methods: Hha I was used to digest genomic DNA, and the digestion efficiency was evaluated by using qPCR technique. Then the optimized MSRE-qPCR method was established. The methylation levels of APC in 45 liver tissues (20 surgically resected HCC specimens and the matched non-cancerous tissues, as well as 5 normal liver tissues) were detected by MSRE-qPCR, and then further validated by using bisulfite sequencing PCR (BSP). The results of MSRE-qPCR were compared with those of methylation-specific PCR (MSP) assay. MSRE-qPCR method was used to detect the APC methylation status of plasma samples from 72 cases of H CC, 37 cases of benign liver diseases and 41 healthy volunteers. Results: The established MSRE-qPCR method could detect as low as 1% methylated target sites in given DNA samples. The results of MSRE-qPCR and MSP showed that APC gene was hypermethylated in HCC tissues. The result of MSRE-qPCR was verified by BSP, and it was comparable with that of MSP (Kappa=0.955, P<.000 1). Methylation level of plasma APC in patients with H CC was significant higher than those in patients with benign liver diseases and the healthy volunteers (P<.000 1).Combined analysis of plasma APC methylation and serum alpha-fetoprotein (AFP) revealed an increased diagnostic efficacy for HCC. Conclusion: MSRE-qPCR is a method for quantitative analysis of APC methylation level. Methylation analysis of plasma APC is a valuable method for the noninvasive diagnosis of HCC. Copyright© 2011 by TUMOR.