Subject(s)
Respiratory Insufficiency , Wakefulness , Humans , Prone Position , Respiratory Insufficiency/therapy , OxygenABSTRACT
Our study aimed to investigate the isoform-specific distribution of 14-3-3 in tongue squamous cell carcinoma (TSCC) and their association with cancer progression, and to further discuss their roles in cancer cell survival. In this study, 42 TSCC specimens and their matched normal para-carcinoma sections were collected. The immunohistochemistry analysis identified that 14-3-3σ and ζ isoforms presented significantly higher expression in cancerous tissues compared with the matched normal tongue tissue sections. 14-3-3ζ expression was associated with tumor T stage, lymph node metastasis and poor prognosis of TSCC. In vitro study revealed that 14-3-3ζ silencing alleviated the proliferation and migration of TSCC cells while promoted cancer cell apoptosis. 14-3-3ζ could bind to and inactivate FOXO3a transcription factor, in turn leading to the movement of the 14-3-3ζ-FOXO3a complex from nucleus to cytoplasm, which was inhibited after 14-3-3ζ silencing. Both 14-3-3ζ and FOXO3a silencing increased caspase 3 and 9 activation, while reduced inner mitochondrial membrane potential. Collectively, 14-3-3ζ may serve as a hallmark and prognostic marker of TSCC. 14-3-3ζ can bind to the FOXO3a transcription factor to promote the export of the complex to the cytoplasm, leading to enhanced proliferation and migration of tongue cancer cells.
Subject(s)
14-3-3 Proteins/genetics , Carcinoma, Squamous Cell/metabolism , Tongue Neoplasms/metabolism , 14-3-3 Proteins/metabolism , Apoptosis , Carcinoma, Squamous Cell/mortality , Carcinoma, Squamous Cell/pathology , Cell Movement , Cell Proliferation , Cell Survival , Disease Progression , Female , Forkhead Box Protein O3/genetics , Forkhead Box Protein O3/metabolism , Gene Knockdown Techniques , Gene Silencing , Humans , Male , Membrane Potential, Mitochondrial , Middle Aged , Prognosis , Protein Transport , RNA, Small Interfering/genetics , Tongue Neoplasms/mortality , Tongue Neoplasms/pathologyABSTRACT
Chicken interleukin 2 (IL-2) is one of important nonmammalian cytokines isolated recently. The influencing of IL-2 on immunogenicity of DNA vaccine was examined using infectious bursal disease virus as a model. The IL-2 cDNA of Xiaoshan chicken and the polyprotein gene of IBDV-ZJ2000 were amplified by RT-PCR, cloned, sequenced and inserted into the control of CMV promoter and enhancer of pCI vector. 14-day-old chickens were vaccinated intramuscularly with DNA vaccine, two weeks later, they were boosted with DNA, and two weeks post boost, they were challenged with virulent IBDV. The results showed that protective responses and neutralization antibody responses of DNA vaccine co-administrated with chicken IL-2 were much higher than those of injected with DNA vaccine alone. Furthermore, the T lymphocyte proliferation response of peripheral blood, thymus and spleen, and the B lymphocyte proliferation response of bursa induced by DNA vaccine can be significantly enhanced by chicken IL-2. These results obviously indicated that chicken IL-2 was a strong adjuvant which can significantly enhance the immunogenicity of IBDV DNA vaccine.
Subject(s)
Infectious bursal disease virus/genetics , Interleukin-2/pharmacology , Polyproteins/genetics , Vaccines, DNA/agonists , Animals , B-Lymphocytes/physiology , Cell Division/drug effects , Cell Division/physiology , Chickens , Drug Synergism , Immunity/drug effects , Infectious bursal disease virus/immunology , Interleukin-2/biosynthesis , Interleukin-2/genetics , Neutralization Tests , Spleen/cytology , T-Lymphocytes/physiology , Thymus Gland/cytology , Vaccines, DNA/administration & dosageABSTRACT
Chicken interleukin 2 (cIL-2) is one of important nonmammalian cytokines isolated recently. In this paper, optimum condition for production of chicken IL-2 in vitro was developed. Isolation of spleen lymphocytes from Xiaoshan chicken, activation by ConA, followed by RT-PCR in a single step, resulted in the synthesis of chicken IL-2 cDNA. The full-length chicken IL-2 cDNA was 737 bp, encoding a 143 amino acids precursor. Only 1--5 amino acid difference were found compared with other three published chicken IL-2s. This IL-2 shared 69.4% homology with turkey IL-2 and shared 21.2%--9.4% homology with mammalian IL-2. The predicted protein had a leader sequence composed of 22 amino acids, and four conversed cysteines allowing the formation of two intrachain disulfide bonds. There were four regions of heptad repeats, with hydrophobic amino acids at positions 1 and 4, were presumably forming amphipathic alpha-helices. These regions were equivalent to mammalian helices A, B, C and D. The amino acids at positions 40(D), 65(Y), 82(E), 108(N) and 142(Q) might play roles in binding to receptors of chicken IL-2. Phylogenetic tree analysis indicated that the chicken IL-2 may have evolutionary relationship with mammalian IL-2 they showed however species difference in function because of selective pressure of immune systems.
