Abstract

Digoxin (DG) is widely used in the treatment of various heart conditions, namely atrial fibrillation, atrial flutter and sometimes heart failure. DG inhibits the growth of cancer cell lines at concentrations commonly found in cardiac patients. Annexin 2 (ANXA2), a calcium-dependent phospholipid-binding protein, is involved in diverse cellular processes such as cell motility, linkage of membrane-associated protein complexes to the actin cytoskeleton, endocytosis, fibrinolysis, ion channel formation, and cell matrix interactions. ANXA2 overexpression is important to maintain the malignancy of cancer cells. The aim of this study is to find the roles of ANXA2 in DG-regulatory hepatocellular carcinoma (HCC) by observing the proliferation, apoptosis, and morphology in DG treatment. Material and Methods:The shRNA is transfected in 293T breast cancer cell line to product the virus cultured suspension, which can make the several HCC cancer cell lines to express ANXA2 or not in virus infection, and they were used to treated with DG ineffective dose. Cells apoptosis was observed by MTT assay, cells containing transforming oncogenes grown in focus-forming assay. Results: 9 liver cancer cell lines were tested by Western blot and found in ANXA2 expression. We choose HA22T due to its higher expression in ANXA2 and characteristic in migration. Moreover, ANXA2 shRNA (6144/6145/6322/9719) were bought by Academia Sinica and used to knockdown HA22T cell line by Transfection and Infection. The shANXA2-6322 and shANXA2-9717 were confirmed in QPCR, RT-PCR and Western blot, and chosen to used in Transwell. We found that whether coating or uncoating, the cell migration and invasion were decreased in shANXA2-6322 and shANXA2-9717 cell line. After treating with DG, the decrease in cell migration and invasion were stronger especially in DG D at dose of 0.1uM . Moreover, the shANXA2-6322 and shANXA2-9717cell survival rate were also decreased under DG treatment in MTT assay. Conclusion: Weather coating or uncoating, the cell migration and invasion were decreased in shANXA2-HA22T. After treating with DG, the decrease in cell migration and invasion were stronger especially in DG at dose of 0. 1uM Moreover, the shANXA2-HA22T cell survival rate were also decreased under DG treatment in MTT assay. Keywords Annexin 2 (ANXA2), shANXA2-HA22T, Digoxin (DG), migration, invasion