AIM: To investigate the e xpression and clinical significance of DEK, cyclin D1, insulin-like growth factor Ⅱ (IGF-Ⅱ),glypican 3 (GPC3), ribosomal phosphoprotein 0 (rpP0) mRNA in hepatocellular carcinoma (HCC) and its paraneoplastic tissues.METHODS: The expression of mRNAs of DEK, cyclin D1,IGF-Ⅱ, GPC3 and rpP0 mRNA was detected in HCC and its paraneoplastic tissues by multiplex RT-PCR.RESULTS: By the simplex RT-PCR, the overexpression of mRNAs of DEK, cyclin D1, IGF-Ⅱ, GPC3, rpP0 mRNA in HCC and its paraneoplastic tissues was 78.1%, 87.5%,87.5%, 75.0%, 81.3% and 15.6%, 40.6%, 37.5%, 21.9%,31.3% respectively (P＜0.05). By the multiplex RT-PCR,at least one of the mRNAs was detected in all HCC samples and in 75.0% of paraneoplastic samples (P＞0.05). However,all these five mRNAs were found in 68.8% of HCC samples,but only in 9.4% of paraneoplastic tissues (P＜0.05). The positive expression of mRNAs of DEK, cyclin D1, IGF-Ⅱ,GPC3, rpP0 in well- and poorly-differentiated HCC was 89.0%, 66.7%, 66.7%, 66.7%, 77.8% and 73.9%, 95.7%,95.7%, 95.7%, 82.6%, respectively (P＞0.05). The expression of these genes in HCCs with α-feto protein (AFP) negative and positive was 90.0%, 80.0%, 90.0%, 90.0%, 90.0% and 72.7%, 86.3%, 77.3%, 90.9%, 68.2% respectively (P＞0.05).CONCLUSION: The expression of DEK,, cyclin D1, IGF-Ⅱ,GPC3, rpP0 mRNA in HCC is much higher in HCC than in its paraneoplastic tissues. Multiplex RT-PCR assay is an effective, sensitive, accurate, and cost-effective diagnostic method of HCC.