Selective blockage of abnormal Wnt pathway inhibits the growth and invasion of hepatocarcinoma cells / 复旦学报（医学版）

ABSTRACT

Objective To construct the recombinant expression vector encoding antisense Tcf fragment for the blockage of abnormal Wnt pathway, and to investigate its effect on the biological behaviors of human hepatocarcinoma cells. Methods Antisense expression vector was transfected into hepatocarcinoma cells SMMC-7721 with GeneJammer. RT-PCR and Western blot were used to detect Tcf expression. Cell proliferation and motility were compared by growth curves and Transwell plate assay. Cell apoptosis was determined by Annexin V and cell cycle was examined by fluorescent staining. Results The stable transfection of antisense Tcf in SMMC-7721 cells significantly reduced Tcf expression at both mRNA and protein levels. Compared with parental and mock-transfected 7721 (7721-vector) cells, antisense Tcf RNA transfected cells 7721-pTas showed much decreased activities of proliferation, migration and invasion in vitro. Furthermore, the apoptosis rate of 7721-pTas cells [(26.34±2.07)%] was significantly higher than that of 7721-vector cells [(6.53±1.02)%] and parental SMMC-7721 cells [(4.33±0.68)%] (P<0.001). The percentages of G0-G1 phase antisense transfected cells were 20.24% and 20.95%, higher than parental SMMC-7721 and 7721-vector cells, and percentages of S phase antisense transfected cells were 11.8% and 11.38%, lower than parental SMMC-7721 and 7721-vector cells, respectively. Conclusions Antisense RNA suppress the growth ability of liver cancer cells by inducing cell apoptosis and impeding the progress of cell cycle, which suggests that selective blockage of abnormal Wnt signal pathway by antisense Tcf RNA may be a potential new gene therapy for liver cancer.