【摘要】 目的建立实时荧光定量PCR(FQ-RT-PCR)检测DHX32 mRNA的方法并研究其在结直肠癌中的表达。方法用Taq-Man探针建立检测DHX32 mRNA的FQ-RT-PCR方法,评价其特异性、重复性及扩增效率,并用该法检测DHX32 mRNA在结直肠癌中的表达水平。结果批内变异系数1.1%～1.9%,批间变异系数1.2%～2.5%;扩增效率为0.85,相关系数为0.9990。癌组织DHX32 mRNA表达水平中位数为1.90×10-3(四分位间距5.47×10-3),明显高于癌旁组织0.09×10-3(四分位间距1.41×10-3),两者差异有统计学意义(P<0.05)。DHX32 mRNA表达水平与结直肠癌癌栓形成、淋巴结转移、组织学分级以及Dukes分期关系密切(P<0.05)。肿瘤的恶性程度越高,DHX32 mRNA表达水平越高。结论FQ-RT-PCR检测DHX32 mRNA的方法特异性好,重复性高,操作简单,无污染。DHX32 mRNA表达上调可能在结直肠癌发生、发展中起重要作用,其表达水平可作为结直肠癌恶性程度的评价指标。更多还原

【Abstract】 Objective To establish real-time fluorescence quantitative polymerase chain reaction (FQ-RT-PCR) method for detection of DHX32 mRNA and investigate DHX32 expression in colorectal carcinoma.Methods Using TaqMan probe, a real-time fluorescence quantitative PCR method was established, and its specificity, reproducibility and amplification efficiency were evaluated. The expression of DHX32 mRNA in colorectal carcinoma tissues and adjacent tissues was determined.Results The intra-assay coefficient variation was from 1.1% to 1.9%.The inter-assay coefficient of variation was from 1.2% to 2.5%.The amplification efficiency was 0.85. The coefficient correlation was 0.990. The expression level of DHX32 mRNA in colorectal carcinoma(M=1.90 ×10-3,QD=5.47×10-3) was significantly higher than that in adjacent colorectal tissue (M=0.09×10-3,QD=1.41×10-3).The different expression of DHX32 in colorectal carcinoma was significantly associated with tumor thrombus, lymphatic follicle diversion, histological grade and DUKES stage (P<0.05).The more malignant, the higher the expression level of DHX32 was.Conclusions FQ-RT-PCR method for determination of DHX32 mRNA is specific, reproductive, simple and pollution-free. The upregulation of DHX32 may play an important role in the tumorigenesis and development of colorectal carcinoma and may be a reference indicator for malignancy of colorectal carcinoma.更多还原