Silencing of LMP2A by Lentivirus-mediated RNAi Inhibits Growth of Epstein-Barr Virus-associated Gastric Carcinoma Cells in vitro / 胃肠病学

ABSTRACT

Background:Epstein-Barr virus (EBV)is associated with various human lymphoid and epithelial malignancies such as Burkitt's lymphoma,nasopharyngeal carcinoma and gastric carcinoma. LMP2A,a virus-encoded latent membrane protein is expressed in a portion of EBV-associated gastric carcinoma (EBVaGC)and has been shown to be related with the tumorigenesis and progression of EBVaGC. Aims:To explore the effect of LMP2A silencing on growth of EBVaGC cells in vitro by using a lentivirus-mediated RNA interference (RNAi)to inhibit LMP2A gene expression. Methods:A lentivirus vector pGCSIL-LMP2A-shRNA-LV and a negative control vector were constructed and transfected into the EBVaGC cell line GT38. Real-time PCR and Western blotting were used to determine the inhibitory effect of the lentivirus vector;CCK-8 assay,colony formation assay and flow cytometry were employed to assess the cell growth,cell cycle and apoptosis of GT38 cells. Results:In GT38 cells transfected with LMP2A-shRNA-LV,the expression level of LMP2A mRNA was decreased by 65. 4%,and that of LMP2A protein was reduced by 50. 8%;the cell proliferation was inhibited,the colony formation ability was suppressed,the percentage of cells in G0 / G1 phase and apoptotic rate were increased when compared with those transfected with negative control vector or without transfection (P < 0. 05). Conclusions:Lentivirus-mediated RNAi is effective for silencing LMP2A gene expression,subsequently inhibiting the growth of EBVaGC cells,inducing G0 / G1 phase arrest and enhancing cell apoptosis in vitro. LMP2A is supposed to be a potential target for gene therapy of EBVaGC.