ABSTRACT
BACKGROUND: The antitumor activity of lymphokine activated killer (LAK) cells immunotherapy is not always effective in all patients, especially when used alone. In this study, we investigated the in vitro antitumor activities of a combination of LAK immunotherapy and gene therapy employing an adenovirus carrying the p53 gene (Ad-p53) in human head and neck squamous cell carcinoma. MATERIALS AND METHODS: The in vitro cytotoxicity of LAK cells was tested in H891 cells infected with or without Ad-p53, and the mRNA expression levels of natural killer group 2D ligands (UL16 binding protein (ULBP) 1 to 5) and tumor necrosis factor (TNF-α) in these cells were measured by real-time reverse transcription polymerase chain reaction. RESULTS: Ad-p53 infection increased the cytotoxicity of LAK cells against H891 cells, and also increased the mRNA expression levels of the ULBPs in H891 cells and TNF-α in the LAK cells. CONCLUSION: The antitumor activities of LAK cells in H891 cells were enhanced by Ad-p53. CONCLUSION: The combinational therapy of LAK immunotherapy and Ad-p53 gene therapy may represent a new paradigm for the treatment of head and neck cancer.
Subject(s)
Carcinoma, Squamous Cell/therapy , Genes, p53 , Genetic Therapy/methods , Head and Neck Neoplasms/therapy , Hypopharyngeal Neoplasms/therapy , Immunotherapy, Adoptive/methods , Killer Cells, Lymphokine-Activated/immunology , Adenoviridae/genetics , Carcinoma, Squamous Cell/genetics , Carcinoma, Squamous Cell/immunology , Cell Line, Tumor , Combined Modality Therapy , Cytotoxicity, Immunologic , GPI-Linked Proteins/biosynthesis , GPI-Linked Proteins/genetics , Head and Neck Neoplasms/genetics , Head and Neck Neoplasms/immunology , Humans , Hypopharyngeal Neoplasms/genetics , Hypopharyngeal Neoplasms/immunology , Intracellular Signaling Peptides and Proteins/biosynthesis , Intracellular Signaling Peptides and Proteins/genetics , RNA, Messenger/biosynthesis , RNA, Messenger/genetics , Squamous Cell Carcinoma of Head and Neck , Tumor Necrosis Factor-alpha/biosynthesis , Tumor Necrosis Factor-alpha/genetics , Tumor Suppressor Protein p53/geneticsABSTRACT
CONCLUSION: Our results indicate that siRNA E6 and/or E7 may have potential as a gene-specific therapy for human papillomavirus (HPV) type 16 (HPV16)-related squamous cell carcinoma of the head and neck (HNSCC). OBJECTIVES: To evaluate the effectiveness of siRNA targeting E6 and/or E7 on the in vitro and in vivo growth suppression of HPV16-related HNSCC. METHODS: HPV16-related HNSCC (UM-SCC47) cell lines were used for the present study. Expression of HPV viral oncogenes E6 and/or E7 and their cellular targets, p53 and pRb, was evaluated by real-time PCR, Western blotting, and immunofluorescence staining. To study the effect of siRNA on tumor growth in vivo, we developed animal models. Representative tumors harvested from each group were processed for apoptosis analyses (TUNEL assay) and immunofluorescence staining for p53 and pRb. RESULTS: E6 and E7 oncogenes of HPV16 were down-regulated by E6 and/or E7 targeting siRNAs, respectively. The expression of p53 and pRb proteins in both the E6 siRNA group and E7 siRNA group was up-regulated compared with those of control groups. The cellular proliferation and apoptosis indexes of E6 and/or E7 siRNA groups were higher than those of controls. In vivo studies showed significant inhibitory effect of E6 and/or E7 siRNA compared with those of control groups, which was consistent with in vitro studies.
