The Impact Of MiRNA Interference Vector Targeting Alpha-1,3 Fucosyltransferase Ⅳ On The Tumor Biological Behaviors Of HepG₂

Background Primary liver cancer was the 6th most common cancer and the 3st leading cause of cancer deaths in the world, and was the 2nd main cause of cancer deaths in China, there were about 600,000 people died of liver cancer worldwide every year. Primary liver cancer not only affected the human life span and the quality of life seriously, but also created a heavy burden on individual family and the national economy[1-3]. Currently, the most effective treatment for the primary liver cancer is still the comprehensive treatments in which surgical resection is the main way, but the metastasis and recurrence of liver cancer after surgery impact on the long-term outcome of patients with liver cancer severely, there were documents indicating that the rate of hepatocarcinoma recurrence rate was 20-64% after 1 year, and up to 57-81% after 3 years.Metastasis and recurrence of cancer was a complex pathological process, and the adhesion between cells had a close ties with the process. The study showed that the adhesion molecules involved in the process of tumor metastasis and recurrence, and played an important role. Some studies of cancers including the colon, breast, prostate cancer and so on showed that the expression of SLeX, cell surface antigen expressed in these cancer, was elevated and correlated with the prognosis significantly[4.5]. In liver cancer tissue, the expression of SLeX was higher than that of the adjacent normal tissues[6]. Similarly, Zhang found that the expression of FUT4 was also increased in the study of hepatocellular carcinoma[7]. The documents indicated that, FUTs took participation in the human physiological and pathological processes, such as body development, angiogenesis, fertilization, cell adhesion, signal conditioning, inflammation, tumor metastasis and so on[8]. According to the research of inflammation and other cancers, we found that the synthesis of SLeX was relevant to FUTs, and that FUTs had diversity and showed a certain difference expression with the temporal and stage[9-13]. However, the relationship between SLeX and FUT4 is vague in HCC, and their roles in the mechanisms of metastasis and recurrence of the liver cancer have not been elaborated in detail. The research on FUT4 and SLeX can help us understand the mechanism of metastasis and recurrence of liver cancer further, and provide a theoretical guidance for the treatment of liver cancer.Objective To investigate the impact of FUT4 on the expression of SLeX on the surface of cell line-HepG₂, and on the tumor biological behaviors of HepG₂, then explore the role of FUT4 in the metastasis of hepatocellular carcinoma.Methods Construct the miRNA interference vector targeting FUT4, transfect the vector into HepG₂ with Lipofectamine, and screen the transfected cells with antibiotics to get the stable transfected cell line; identity the interference efficiency of miRNA vector for FUT4 with FQ-PCR and Western-blot; compare the difference expressions of SLeX between the untransfected cells and the transfected cells and the differences of migration, invasion and adhesion abilities between the untransfected cells and transfected cells.Results The mRNA and protein of FUT4 was significantly decreased in hepatocarcinoma cell line after miRNA interference, the expression of SLeX on the surface of transfected cells was reduced correspondingly, and the migration, invasion and adhesion abilities of cell lines-HepG₂ were inhibited to some extent in the transfected cells.Conclusions In hepatocacinoma cell line-HepG₂, FUT4 takes participation in the synthesis of SLeX partly and has impact on the tumor biological behaviors of cell line HepG₂. Therefore, FUT4 plays a role in the mechanism in the metastasis of hepatocarcinoma.