The Study Of MHBs~t Gene Mutation And Its Significance In Peripheral Blood Of Hepatocellular Carcinoma

【Objective】The purpose of this study was to detect gene mutation in Truncated middle surfaceprotein (MHBst)of patients with HCC、HBC and CHB, make sure the types of MHBstgene mutation in HCC patients, explore mechanism of the HCC occurrence anddevelopment and its significance, and design a simple laboratory method for thedetected of MHBstgene mutation.【Methods】Genomic DNA was extracted from plasma sample of96patients with HCC、40patients with HBC and51patients with CHB respectively. MHBstgene was amplifiedby polymerase chain reaction (PCR), followed by DNA sequencing to detect genemutation; compare the rate of MHBstgene mutation between HCC group andnon-HCC (HBC、CHB) group in peripheral blood; compare the correlation betweenMHBstgene status and clinical pathological characteristic、 NF-κB in HCC group;The detected gene mutation was further tested by TaqMan-MGB probe-basedreal-time fluorescent quantitative PCR.【Results】1.Two MHBstgene mutations out of187patients were identified, which novel pointmutation was MHBst165(C>A) and reported point mutations was MHBst78(G>T)respectively.2.Compared with non-HCC group, HCC group（55/96，57.3%） have a higher rate ofMHBstgene mutation than the non-HCC group（37/91，40.7%）P=0.023;There wereno significant correlation between MHBstgene mutation and clinical pathologicalcharacteristic in cases with HCC, The expression of NF-κB in HCC cases with MHBstgene mutation was higher than that in cases without MHBstgene mutation in HCCtissue. 4.TaqMan-MGB probe method was successfully designed to detect point mutationMHBst165(C>A).【Conclusions】1. A novel point mutation MHBst165（C>A）was found in our research.2.The positive rate of MHBstof gene mutation is higher in patients with HCC.MHBstmight have a function of transactivation in patients with HCC through up-regualtingthe expression of NF-κB.3.TaqMan-MGB probe assay might contribute to mass screening of MHBst165（C>A）with its easy and quick operation.