Expression Of Co - Associated Protein ANXA7 / Gal-3 / SRI In Mouse Hepatocarcinoma Cells And Its Effect On Cell Biological Behavior

Objective To study the expression, subcellular location and roles of annexin A7/galectin-3/sorcin in mouse hepatocarcinoma ascites cell lines with different metastaticpotentials in vitro, for further discuss its roles in tumor metastasis.Methods Mouse hepatocarcinoma cell lines Hca-F with high lymphatic metatasticpotential and Hca-P with low lymphatic metastatic potential were used. qRT-PCR wasperformed to analyze the level of gene expression differences in Hca-F/Hca-P; Westernblot was performed to detect the level of protein expression difference in Hca-F/Hca-P;Immunofluorescence assay was used to detecting the localization of annexin A7/galectin-3/sorcin.Results The gene level and protein level expression of annexin A7in Hca-F wererespectively3.13folds and1.31folds higher than that in Hca-P cells the gene level andprotein level expression of galectin-3in Hca-F were respectively1.56folds and1.35folds higher than that in Hca-P cells; the gene level and protein level expression ofsorcin in Hca-F were respectively1.5folds and1.8folds higher than that in Hca-P cells.Annexin A7had two kinds of protein subtypes:51kDa and47kDa, The47kDaannexin A7expressed lots than51kDa annexin A7, the ODs of47kDa annexin A7versus51kDa annexin A7in Hca-P cells was45±5versus174±12. The proliferation,migration and invasion ability of Hca-F cells were significantly higher than Hca-P cellsin vitro. Immunofluorescence assay indicate annexin A7/galectin-3/sorcin mainlyexpressed in cytoplasm.Conclusions The expressions of annexin A7and its binding partner galectin-3/sorcinconsist with the biology ability of mouse hepatocelluar carcinoma cells, they may playimportant roles in the process of tumor proliferation, migration, invasion, and lymph metastases of mouse hepatocelluar carcinoma, and lay the foundations for the furtherexplore its mechanism and function. Objective To study the influences of annexin A7on the expression of galectin-3inhepatocellular carcinoma and its biology activity in vitro, for further discuss its roles intumor metastasis.Methods Mouse hepatocarcinoma cell lines Hca-P/PANXA7-control/PANXA7-downwere used.qRT-PCR was performed to analyze the level of gene expression differences in Hca-P/PANXA7-control/PANXA7-down; Western blot was performed to detect the level of proteinexpression difference in Hca-P/PANXA7-control/PANXA7-down; Immunofluorescence assaywas used to detecting the localization of annexin A7and galectin-3; Cell proliferationwas assessed by cell counting kit-8; the Boyden-transwell assay was performed toanalyze cell migration ability and cell invasion ability; IPA and genego (metacore)software was used to analyze the roles of galectin-3and annexin a7in network andpathway.Results Annexin A7was successfully down-regulated, the mRNA level and proteinlevels of galectin-3were decreased, the proliferation, migration and invasion ability ofHca-P cells were significantly suppressed. immunofluorescence assay indicate annexinA7and galectin-3were co-located. Annexin A7and galectin-3played roles in DNAdamage and cell proliferation cycle checkpoint arrest pathway.Conclusions The interaction of annexin A7and its binding partner galectin-3may playimportant roles in the process of tumor proliferation, migration, invasion and lymphmetastases of mouse hepatocelluar carcinoma.