| Objective:The hepatocellular carcinoma(HCC) is one of the highest incidence malignant tumor in digestive system. Respecting of the high incidence of hepatitis in our country, the morbidity and mortality of hepatoma have the elevated tendency year by year in recent years. At present, there are no efficacious therapia to cure hepatoma, because it has the bionomics of the powerful infiltration, easy recrudescence and metastasis, and so on. It has become the oncological hot topic to research the etiopathogenisis, pathogenesy, bionomics and original therapeutics of hepatocellular carcinoma. Heparanase (hpa), which expresses commonly in malignant tumors, is essential in activating angiogenesis for the proliferation and migration of tumor cells. TNF-αand IL-1β, which have the biological functions of immunological regulation and antineoplastic effect, are the most important and typical cytokines secreted by several kind of immunological cells. Recently it has been reported that endothelial heparanase can be highly induced by inflammatory cytokines, such as TNF-αand IL-1β, in inflammatory diseases. Whether this effect in tumor takes place the way equally did not appear in literature. In this research, we explored whether heparanase can be up-regulated in hepatocellular carcinoma cells SMMC-7721 treated with TNF-αand IL-1βrespectively. By detecting the expression of Hpa mRNA and protein in SMMC-7721 cultured in normal and treated conditions by RT-PCR and Western blot, we investigate the effect of TNF-α,IL-1βon the expression of heparanase. It will provide a theoretical foundation for clarifying the mechanism of invasion and metastasis of HCC and exploring a new therapeutic way.Methods: SMMC-7721 cells were divided as control group and experiment group. The former were cultured with normal condition, the later were treated by TNF-αor IL-1βwith various concentration and time. All of the cells were collected on time. The expression of Hpa mRNA level in cultured SMMC-7721 was evaluated by RT-PCR. The Hpa protein level in cytoplasm was evaluated by western blot. The OD value of electrophoresis strip of DNA product or that of Hpa protein was analyzed by gel scanner. Beta-actin was used as an internal standard. The relative expression level of Hpa was represented with the ratio between produces of Hpa and that of beta-actin. Data were analyzed with ANOVA and T test using SPSS12.0 statistical software. A level of P<0.05 was considered statistically significant.Results: 1 At the same time, following the increasing of the concentration of TNF-αor IL-1β, expression of Hpa mRNA was up-regulated gradually. The effect was dose-dependent at the concentration of 106U/L or lower for TNF-αand 5.0μg/L or lower for IL-1β. Furthermore, the effect of promotion was close to peak when the TNF-αconcentration up to 106U/L or the IL-1βconcentration up to 5.0μg/L, but no time-dependent pattern observed.2 The Hpa protein level in cytoplasm was elevated gradually with the concentration increasing, too. It's tendency is same to that of Hpa mRNA following the increasing of the concentration of TNF-αor IL-1β. Both the dose-dependent pattern and the time-dependent pattern were observed.Conclutions: 1 TNF-αand IL-1βcan promote the expression of Hpa at level of mRNA and protein in hepatic cancer cell line, and the effect of promotion was dose-dependent. Furthermore, time-dependent pattern were observed at the level of Hpa protein.2 Low concentration of TNF-αor IL-1βcan not resist invasion and metastasis of tumor,on the contrary, it will promotes the secretion of Hpa,and then promote to degrade the extracellular matrix and vascular basement membrane and induces angiogenesis to promotes invasion and metastasis of tumor. |
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