The Effect Of The Expression Of Annexin A7 On Lymphatic Metastasis Of Mouse Hepatocarcinoma

Objective:Hepatocarcinoma is one of common malignant tumors, featured as high invasion,early metastasis,high recurrence after operative or interventional treatment and poor consequences.Early detection and prevention of hepatocarcinoma metastasis is very important to improve the prognosis.Thus,we wish to explore an appropriate approach to understand the mechanism of metastasis and proper treatment in hepatocarcinoma. Recently,researchers all over the world have done many works on Annexin A7,which found that abnormal expression of Annexin A7 were correlated with the development among breast cancer,prostate cancer and invasive phenotype of malignant melanoma.some studies also found that Annexin A7 involved in invasion and metastasis of tumor cells,so further studies about Annexin A7 which focused on molecular mechanism and signal pathway of lymphatic metastasis were very important to improve the prognosis of hepatocarcinoma patients.Methods:We focused on the following six kinds of cells:Hca-F, Hca-P, Hca-F-pSilencer,Hca-F-shRNA, Hca-P-pcDNA3.1 and Hca-P-Annexin A7, and made a model of lymphatic metastases via injecting theses cells mentioned above into the foot pads of 615 rats.Lymphatic metastases rates and proteins changes in primary tumor and metastasized lymph nodes were detected by HE stain,immmunocytochemistry and western blot after down and up regulating the expression of Annexin A7 gene.Results:1. Compared with Hca-F-pSilencer cell group,lymphatic metastases rates were relatively higher in Hca-F-shRNA cell group(P=0.018). Compared with Hca-P-pcDNA3.1 cell group, lymphatic metastases rates were significantly decreasing in Hca-P-Annexin A7 cell group,(P=0.005), the differences between the two groups were notable significant.2.1)The expression tendency of Annexin A7 in primeval tumor cell of Hca-F-shRNA cell group were lower than Hca-F-pSilencer cell group (P=0.000),the differences between the two groups were notable significant; the expression of Annexin A7 in primeval tumor cell of Hca-P-Annexin A7 cell group were higher than Hca-P-pcDNA3.1 cell group(P=0.000),the differences between the two groups were notable significant.2)The expression of Annexin A7 in metastasized lymph nodes of Hca-F-shRNA cell group were higher than Hca-F-pSilencer cell group (P=0.002), the differences between the two groups were notable significant.3)The expression of Annexin A7 in primeval tumor cell of Hca-F-pSilencer cell group were higher than in metastasized lymph nodes (P=0.001),the differences between the two groups were notable significant. The expression of Annexin A7 in metastasized lymph nodes of Hca-F-shRNA cell group were higher than in primeval tumor cell (P=0.000), the differences between the two groups were notable significant. The expression of Annexin A7 in metastasized lymph nodes of Hca-P-pcDNA3.1 cell group were higher than in primeval tumor cell (P=0.001),the differences between the two groups were notable significant.3. In Hca-F-pSilencer cell group and Hca-F-shRNA cell group, the expression of Annexin A7 in primeval tumor cell mainly located in cytosol, partially located in membrane,while the expression of Annexin A7 in metastasized lymph nodes was only located in cytosol; In Hca-P-pcDNA3.1 cell group and Hca-P-Annexin A7 cell group, the expression of Annexin A7 in primeval tumor cell was mainly located in cytosol, partially located in nucleus, in metastasized lymph nodes the expression of Annexin A7only located in cytosol too.4. In Hca-F cell with a high metastases potential, the expression tendency of 51KD and 47KD are consistent with total protein,charactered as high-lower-high.In Hca-P cell with a low metastases potential,only 47KD are consistent with total protein charactered as decreasing gradually; the expression of total protein and 47KD in F-shRNA28 cell group are lower than F28 cell group, while, the expression of 51KD in F-shRNA28 cell group are higher than F28 cell group; the expression of 51KD and total protein in PA728 cell group are higher than P28 cell group,but the expression of 47KD are lower than P28 cell group.Conclusion:1. After down regulating Annexin A7 expressed in Hca-F cell, its' lymphatic metastases rates increased in vivo.After up regulating Annexin A7 expressed in Hca-P cell, its'lymphatic metastases rates decreased in vivo.2. After down and up regulating the expression of Annexin A7 gene, Annexin A7 protein have different expression in primeval tumor cell and metastasized lymphatic node which indicated that Annexin A7 gene might play an importent role in the process of tumor lymph node metastases.3. The expression of Annexin A7 in Hca-F cell and Hca-P cell was significant different location,which indicated that Annexin A7 had close relationship with the development of hepatocarcinoma.4. The two subtypes of Annexin A7,51KD and 47KD which had different tendency of expression were related to the progression of hepatocarcinoma.