Abstract:Objective:To quantitatively detect the expression level of RUNX3 in human primary hepatocellular carcinoma (HCC) by real-time fluorescent quantitative RT-PCR, and assess its relationship with the clinicopathological features of the disease.
Methods:Total RNA isolated from samples, including sixty-five HCC, adjacent liver tissues, six human liver cancer cell lines and a human normal liver cell line, were reversed into cDNA. Real-time fluorescent quantitative RT-PCR method was used to analyze the expression level of RUNX3 gene. All the quantitative results were confirmed by Western blotting.
Results:The expression of RUNX3 mRNA was 4.2-fold lower in HCC tissues than in adjacent liver tissues (P<0.001), 8-fold lower in 18 HCC tissues,4-fold lower in 21 HCC tissues and 2-fold lower in 3 HCC tissues. A signicficant down-regulation of RUNX3 mRNA was found in 66.67%(4/6)of liver tumor cell lines. Furthermore, Western blot analysis showed that there was also significant difference in RUNX3 protein expression between the HCC and adjacent liver tissues, which was in accordant with the results of quantitative RT-PCR analysis of RUNX3 mRNA levels. Lower expression showed significant correlation to cirrhosis (P=0.028) and histologic type of HCC (P=0.000).
Conclusions:The expression of RUNX3 is decreased in HCC. The RUNX3 gene may play an important role in the development of HCC.