1. Enhanced T cell immunity by B7-H4 downregulation in nonsmall-cell lung cancer cell lines
- Author
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Jiang Cg, Huang Pl, Siqing Sun, Jin Yl, and Yong Lin
- Subjects
Lung Neoplasms ,T cell ,T-Lymphocytes ,Down-Regulation ,Apoptosis ,Biochemistry ,Jurkat cells ,Interferon-gamma ,Jurkat Cells ,Immune system ,Carcinoma, Non-Small-Cell Lung ,Cell Line, Tumor ,medicine ,Humans ,RNA, Messenger ,RNA, Small Interfering ,Cell Proliferation ,A549 cell ,Immunity, Cellular ,Cell growth ,business.industry ,Biochemistry (medical) ,Cell Cycle ,Cell Biology ,General Medicine ,Cell cycle ,V-Set Domain-Containing T-Cell Activation Inhibitor 1 ,Coculture Techniques ,Interleukin-10 ,Gene Expression Regulation, Neoplastic ,medicine.anatomical_structure ,Cell culture ,Cancer research ,Cytokines ,Interleukin-2 ,RNA Interference ,business - Abstract
OBJECTIVES: T cell immunity plays a critical role in host immune surveillance of tumour cell growth and metastatic spread. This study used small hairpin (sh)RNA-mediated gene silencing to target VTCN1 (B7-H4) expression in a nonsmall-cell lung cancer (NSCLC) cell line (A549) and evaluated the effects on T cell immune activity using an in vitro coculture system. METHODS: VTCN1-specific shRNA-expressing plasmid was transfected into A549 cells. Mock transfected and empty plasmid-transfected A549 cells served as controls. VTCN1 expression in A549 cells was determined by reverse transcription– polymerase chain reaction (RT—PCR) for VTCN1 mRNA and Western blotting for B7-H4 protein. Transfected A549 cells were cocultured with Jurkat cells. Jurkat cells were examined for proliferation, apoptosis, cell cycle distribution and intracellular cytokine mRNA and protein levels. RESULTS: VTCN1-specific shRNA efficiently knocked down VTCN1 mRNA and B7-H4 protein levels in A549 cells. This downregulation led to enhanced Jurkat cell proliferation, decreased apoptosis, stimulated cell cycle progression and elevated production of interferon-γ, interleukin (IL)-10 and IL-2. CONCLUSIONS: B7-H4 negatively regulates T cell-mediated antitumour immunity in NSCLC.
- Published
- 2012