Effect Of Fgl2 Prothrombinase On Growth Of Tumor And Neovascularization In Hepatocellular Carcinoma

Background and objective:Nowadays, malignancy is one of the most common diseases seriously imperiling human beings. In china, the annul number of cancer incidence cases was about 1.6 million. The study on carcinoma is becoming an issue increasingly concerned by scientists all over the world. The association between thrombosis and cancer was observed by Professor Armand Trousseau in 1865. Thrombin was now believed one of the key mediators of this link. We and many others have described a new procoagulant: fgl2 prothrombinase, a member of the fibrinogen superfamily with serine protease actively, which directly cleaves prothrombin to thrombin which increased the growth and metastatic potential of tumor cells, thereby, initiating a cascade coagulating reaction. Our previous research showed that fgl2 prothrombinase was expressed in malignant tumor tissues including colon, breast, lung, gastric, cervical, and esophageal tissues from patients. The hfgl2 expression was closely associated with local fibrin deposition. In this study, we aim to investigate the role of fgl2 in tumor growth by establishing the HCC nude mouse model and to detect the differential expression of fg12 prothrombinase in nude mice subcutaneous transplanting tumor among three groups. The effect of hfg12 on the tumor growth and neovascularization provides a new target for gene treatment of liver cancer. Method: pcDNA6.2-fgl2-miRNA was transfected into human hepatoma cell line HCCLM6. Then we established HCC nude mice model by subcutaneous injection of human HCCLM6,HCCLM6-noncorrelation and HCCLM6-fgl2(-)into the dorsal scapular skin of nude mice. Dynamic observation lasted for 42 days, tumor growth size, rate of tumor formation, latent period and its blood vessel rebirth were compared among HCCLM6 group ,HCCLM6-noncorrelation group and HCCLM6-fgl2(-)group. Then the nude mice were sacrificed and the tumors were removed. The immunoenzymatic staining was applied to assess the difference of fg12 expression and MVD in nude mice subcutaneous transplanting tumor tissue.Results:The HCC nude mouse model was successfully establised. Observation of the nude mice subcutaneous transplanting tumor showed that the incubation period and the formation rate of the tumors in HCCLM6-fgl2(-)group were longer and greater than HCCLM6 group and HCCLM6-noncorrelation group, the size of the transplanting tumor in HCCLM6-fgl2(-) group was smaller than the HCCLM6 group and HCCLM6-noncorrelation group. The immunoenzymatic staining of fgl2 and CD34 showed that the expression of fgl2 in tumor epithelial cells in HCCLM6 group was not apparent and MVD(microvessel density) in HCCLM6-fgl2(-)group was less than HCCLM6 group and HCCLM6-noncorrelation group.Conclusions:HCC nude mouse model was built successfully. Dynamic observation of HCC nude mice model showed that HCCLM6 cell after fgl2 silence could inhibit the tumor growth significantly. The immunoenzymatic staining suggested that HCCLM6 cell after fgl2 silence suppressed neovascularization and fgl2 expression in vivo. The results above explained that fgl2 might influence the biological behavior of malignant tumors by inducing angiogenesis through cruor-dependent and non cruor-dependent ways. Thus fg12 promoting the tumor growth and the neovascularization is one of the important links of pathology and physiology change of tumor growth and transfer.