Initial Study On The Expression Of Spp-1, Gdf-15 And Cxcl-1 In Human Small Hepatocellular Carcinoma

Primary liver cancer is the fifth most common malignancy worldwide, which about half of cases are from China. The long-term prognosis of patients undergoing potentially curative hepatic resection is still poor, with reported 5-year survival rates ranging from 17% to 53%. Despite resection with curative intent, the clinical course is variable and recurrence occurs in a high proportion of cases. The poor prognosis of liver cancer is majorly due to patients'no response to almost all conventional chemotherapy and over 80% patients diagnosed at advanced stage and lost the opportunity for curative resection.The common validated serology tumor-marker in current use for diagnosis of HCC is AFP, which can be detected in the serum of more than 56.1% of cases with HCC. However, AFP is thought to be robust only in following the progression of the disease, but is not sensitive enough as an early stage diagnostic marker. Thus, there is an imperative need for additional diagnostic markers for this disease. Microarray technology allows researchers to simultaneously monitor the expression of thousands of genes, which provides molecular biomarkers to differentiate normal cells from malignant ones.Our group used Affymetrix Rat Genenome 230.2.0 GeneChip to detect the gene expression profile in early stage malignant liver tissues. Furthermore, web-based bio-information and sequence analysis were applied to screen target genes which significantly elevated expressed in tumor tissues and encode secreted proteins.The expression of several genes that encode secreted protein were confirmed at transcript and protein expression level. In this dissertation, we quantitatively validate the expression of SPP-1, GDF-15 and CXCL-1 which are not reported in small hepatocellular carcinoma by real-time fluorescence quantitative PCR and double antibody sandwich ELISA and investigate the clinical value.Methods1,The results of Rat Expression Array 230 2.0 were analyzed by bioinformatics methods. Web-based bio-information and sequence analysis were used to screen target genes which significantly elevated expressed in tumor tissues and encode secreted proteins.2,The mRNA expression of SPP-1, GDF-15 and CXCL-1 was determined by real-time PCR in samples from 15 small HCCs, including tumor tissues and their matched normal liver tissues.3,Protein expression was surveyed by double antibody sandwich ELISA in serum samples, including 15 cases of corresponding tumor patients and randomly healthy adults.Results1,14 candidate diagnostic markers were screened which may encode secreted proteins.SPP-1, GDF-15 and CXCL-1 are three of them which are not reported to have relationship with early HCC.2,The mRNA expression of SPP-1, GDF-15 and CXCL-1 significantly increased in small HCC tissues compared to the paired normal tissues (P<0.05);3,Double antibody sandwich ELISA analysis revealed that GDF-15 protein was expressed higher in the serum in small HCC than in the control group (P<0.05) and the difference was not significant in SPP-1(P>0.05); Protein expression of CXCL-1 revealed the opposite expression pattern of in the serum in small HCC (P<0.05).Conclusions NF-κB signal pathway gene(CXCL-1), P53 signal pathway gene (GDF-15) and SPP-1 may be associated with the hepatocarcinogenesis ; The current results suggested CXCL-1 and GDF-15 should be a potential tumor marker in detecting HCCs at early stage.