The Relationship Between Over-expression Of Annexin A7and Proliferation, Migration And Invasion Of Mouse Hepatoma Cells

Objective:Hepatocarcinoma is one of common malignant tumors.According tothe statistics,about110000people died of liver cancer each year, the mortality rate ofmalignant tumors in third in the digestive system,accounted for about45%of livercancer deaths in world, the clinical and pathological features of hepatocarcinoma arehigh invasivenes metastastic potential recurrence rate and poor prognosis.As its highmetastic potential is the basic reason of its high mortality rate and poor prognosis,it’svery important to make sure the tumor metastic mechanism and explore the effectivetreatment method to improve the prognosis of patients with liver cancer and reducemortality.Recently,many researchers found that abnormal expression of Annexin A7were correlated with the development among breast cancer,prostate cancer andinvasive phenotype of malignant melanoma.This suggests that Annexin A7mayattend to the process of tumor lymphatic metastasis.Hca-F and Hca-P are a pair of syngenetic mouse hepatocarcinoma ascites celllines which have different potential of lymphatic metastasis. Hca-F has highmetastatic rate more than70%,and Hca-P has low metastatic rate less than30%.Inour previous studies,we found that Annexin A7gene express higher in Hca-F thanHca-P.Reducing the Annexin A7expression level can inhibit the proliferation、promote apoptosis,reduce migration and invasion capacities of mouse liver cancercells. Indicately, Annexin A7is one of the key factors which decide lymphaticmetastasize in Hca-F cell.Methods:Constructing PG-CMV-EGFP-Kan/neo-Annexin A7expressionvector and PG-CMV-EGFP-Kan/neo-negative control expression vector,and transfectthe vector into Hca-F cell.Fanally,we gain stable transfection cells which is the overexpression of Annexin A7on Hca-F cells. qRT-PCR and Western blot are used to detect the expressiong of Annexin A7in both mRNA and protein levels.Cell countingkit-8(CCK8) and Transwell are used to compare the ability of multiplication、migration and invasion between the over expression of Annexin A7on Hca-F cellsand the nomal Hca-F cells.Results:1.The Restructuring plasmid gene sequence of synthesized and Annexin A7gene sequences completely consistent,which indicates the restructuring of theAnnexin A7genes plasmid is succeed.The stable transfection cells with overexpressed Annexin A7have been detected by qRT-PCR detection.2. The result of Western blot test is Annexin A7protein in stable transfectioncells are higher than the normal Hca-F cells and the stable transfect NC cells.3. Cell counting kit-8(CCK8) test result is the latest cell proliferation capacityincrease apparently with Annexin A7gene over expressed.4. Transwell test result is the cells migrating ability and the ability of invasionare enhanced with Annexin A7gene over expressed.Conclusion:1. Constructing PG-CMV-EGFP-Kan/neo-Annexin A7expression vector andPG-CMV-EGFP-Kan/neo-negative control expression vector, and transfect intoHca-F cell.Fanally,we gain stable transfection cells with the over expression ofAnnexin A7on Hca-F cells successful.2. The over expressed Annexin A7is proved to enhance the cell proliferation ofHca-F cells,enhance the migration and invation capacities of Hca-F cells.