OBJECTIVETo evaluate the application of DNA vector-based RNA interference for silencing LMP-1 expression in EBV-positive nasopharyngeal cancer (NPC) C611 cells and analyze the inhibiting effects of LMP-1 silencing on metastatic property of NPC cells.

METHODSDNA vectors with LMP-1 shRNA template were transfected into C611 cells. After G418 selection, a LMP-1 silenced sub-clone named C611L was isolated. Effect of LMP-1 inhibition on its metastatic properties was evaluated by cell adhesion, invasion and wound healing assays.

RESULTSshRNA vector mediated stable RNAi against LMP-1 in C611 cells. Loss of LMP-1 significantly altered cell motility, substratum adhesion and transmembrane invasion ability.

CONCLUSIONStable suppression of LMP-1 induced by shRNA vector can inhibit metastatic properties of NPC cells.

Cadherins ; metabolism ; Cell Adhesion ; Cell Line, Tumor ; Cell Movement ; Genetic Vectors ; Herpesvirus 4, Human ; genetics ; growth & development ; Humans ; Nasopharyngeal Neoplasms ; metabolism ; pathology ; virology ; Neoplasm Metastasis ; RNA Interference ; RNA, Small Interfering ; genetics ; Transfection ; Viral Matrix Proteins ; genetics ; physiology

Nasopharyngeal carcinoma(NPC) is a metastatic carcinoma that is highly prevalent in Southeast Asia. Our laboratory has previously demonstrated that the C-terminal 27-kDa polypeptide of human telomerase reverse transcriptase (hTERTC27) inhibits the growth and tumorigenicity of human glioblastoma and melanoma cells. In this study, we investigated the antitumor effect of hTERTC27 in human C666-1 NPC cells xenografted in a nude mouse model. A cocktail of vectors comprising recombinant adeno-associated virus (rAAV) and recombinant adenovirus (rAdv) that each carry hTERTC27 (rAAV-hTERTC27 and rAdv-hTERTC27; the cocktail was abbreviated to rAAV/rAdv-hTERTC27) was more effective than either rAAV-hTERTC27 or rAdv-hTERTC27 alone in inhibiting the growth of C666-1 NPC xenografts. Furthermore, we established three tumors on each mouse and injected rAAV/rAdv-hTERTC27 into one tumor per mouse. Although hTERTC27 expression could only be detected in the injected tumors, reduced tumor growth was observed in the injected tumor as well as the uninjected tumors, demonstrating that the vector cocktail could provoke an antitumor effect on distant, metastasized tumors. Further studies showed the observed antitumor effects included inducing necrosis and apoptosis and reducing microvessel density. Together, our data suggest that the rAAV/rAdv-hTERTC27 cocktail can potently inhibit NPC tumor growth in both local and metastasized tumors and should be further developed as a novel gene therapy strategy for NPC.
Adenoviridae ; genetics ; Animals ; Apoptosis ; Carcinoma ; Cell Line, Tumor ; Dependovirus ; genetics ; Genetic Therapy ; methods ; Genetic Vectors ; Green Fluorescent Proteins ; metabolism ; Humans ; Male ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Microvessels ; Nasopharyngeal Neoplasms ; metabolism ; pathology ; Neoplasm Transplantation ; Recombinant Proteins ; genetics ; metabolism ; Telomerase ; genetics ; metabolism ; Tumor Burden