The Study On Liver Cancer Cells Growth In Nude Mice Model Treated By A Combination Of Sorafenib And 5-FU

Purpose:In this experiment human liver cancer nude mice model are treated with Sorafenib and/or 5-Fu. We quantify the expression of pERK and drug resistant protein like P-gp and TOP2A to testify these two agents’complementary and cooperation in tumor inhibiting which can provide the experimental evidences for clinical use.Methods:Make nude mice liver cancer model using human high metastasis liver cancer cell line (HCCLM3 cell). Treat HCCLM3 cells and nude mice models with negative control, Sorafenib,5-Fu and combination(Sorafenib and 5-Fu). Access the treat response through tumor volume and size measuring and make clear the complementary and cooperation between Sorafenib and 5-Fu by the expression of pERK, P-gp and TOP2A in cells through immunohistochemistry, westernblot and immunocytochemistry.Results:1. In nude mice experiments, the size, volume and lung metastasis number in single Sorafenib group and combination group (combine Sorafenib and 5-Fu) significantly reduce (P<0.05) compare to negative control. But the two groups, single Sorafenib and combination, have no difference (P>0.05).2. In cell experiments, the expression of pERK, P-gp and TOP2A in single Sorafenib group and combination group significantly reduce (P<0.05) compare to negative control. The two groups, single Sorafenib and combination, have no difference (P>0.05).3. In tumor tissue, the expressions of pERK and P-gp in single Sorafenib group and combination group significantly reduce (P<0.05) compare to negative control. The expression of TOP2A in single Sorafenib group significantly reduces (P<0.05) compare to negative control. The expressions of three proteins have no difference between combination group and single Sorafenib group (P>0.05)Conclusion:1. Sorafenib remarkably inhibits the tumor growth and lung metastasis in nude mice model2. Sorafenib remarkably inhibits the expression of pERK in tissue and HCCLM3 cells.3. Sorafenib remarkably inhibits the expression of resistant protein P-gp and TOP2A in tissue and HCCLM3 cells.4. Single 5-Fu can’t inhibits the tumor growth and lung metastasis in nude mice model and expression of pERK, P-gp and TOP2A in tissue and HCCLM3 cells.5. Combination treatment (Sorafenib and 5-Fu) remarkably inhibits the tumor growth and lung metastasis in nude mice model and expression of pERK, P-gp in tumor, it remarkably inhibits the expression of pERK, resistant protein P-gp and TOP2A in HCCLM3 cells.Focus on these former indexes combination treatment (Sorafenib+ 5-Fu) has no further advantage compare to use Sorafenib alone which means the inhibitions come from Sorafenib.6. What kinds of pathways make the low expression of P-gp and TOP2A in Sorafenib treated tissue, direct function or Raf/MEK/ERK signal pathway, are not certain and need further explore?7. Sorafenib can inhibit the expression of resistant protein P-gp and TOP2A in tissue. We still need more experiments to prove that the longer exposure time of cancer tissue in Sorafenib and 5-Fu can give 5-Fu more chance to kill the cancer cells and get better response because of lower 5-Fu resistance.