ABSTRACT
<p><b>OBJECTIVE</b>To explore correlation of seven apoptosis-related proteins (Hsp90a, p53, MDM2, Bcl-2, Bax, Cytochrome C, and Cleaved caspase3) with clinical outcomes of ALK+ anaplastic large-cell lymphoma (ALCL).</p><p><b>METHODS</b>Using immunohistochemistry and immunofluorescence double staining methods, the expressions of these seven apoptosis-associated proteins were studied to clarify their relationship with clinical outcomes of 36 ALK+ and 25 ALK-systemic ALCL patients enrolled between 1996 and 2006. The relationship of these apoptosis-regulating proteins with NPM-ALK status was also evaluated with the tyrosine inhibitor herbimycin A (HA) in vitro by immunocytochemistry, Western blotting and flow cytometric assays.</p><p><b>RESULTS</b>The presence of Hsp90α-, MDM2-, Bax-, Cytochrome C, and Cleaved caspase3-positive tumor cells was found significantly different in ALK+ and ALK-ALCLs, which was correlated with highly favorable clinical outcome. The Bcl-2- and p53-positive tumor cells were found in groups of patients with unfavorable prognosis. Inhibition of NPM-ALK by HA could reactivate the p53 protein and subsequent apoptosis-related proteins and therefore induced apoptosis in ALK+ ALCL cells.</p><p><b>CONCLUSION</b>Our results suggest that these seven proteins might be involved in apoptosis regulation and associated with clinical outcome of ALK+ systemic ALCLs. We also reveal a dynamic chain relation that NPM-ALK regulates p53 expression and subsequent apoptosis cascade in ALK+ ALCLs.</p>
Subject(s)
Adolescent , Adult , Aged , Child , Child, Preschool , Female , Humans , Male , Middle Aged , Young Adult , Apoptosis , Apoptosis Regulatory Proteins , Metabolism , Benzoquinones , Pharmacology , Biomarkers, Tumor , Metabolism , Blotting, Western , Cell Culture Techniques , Cell Survival , Disease-Free Survival , Enzyme Inhibitors , Pharmacology , Flow Cytometry , Immunohistochemistry , Kaplan-Meier Estimate , Lactams, Macrocyclic , Pharmacology , Lymphoma, Large-Cell, Anaplastic , Metabolism , Pathology , Microscopy, Fluorescence , Neoplasm Staging , Prognosis , Protein-Tyrosine Kinases , Metabolism , Receptor Protein-Tyrosine Kinases , Metabolism , Retrospective Studies , RifabutinABSTRACT
This study was aimed to investigate the relationship between immunophenotype of the background lymphocytes and histological subtype of Hodgkin's lymphoma (HL), and its significance. The relative protein expressions of background lymphocytes were detected in 37 HL specimens on the basis of instant-rapid MaxVision(TM) immunohistochemical method and assessed quantitatively with image analysis software IPP6.0. The adoptive antibody included anti-CD3/CD45RO, anti-CD20/CD79a, anti-CD4, anti-CD8, anti-GrB, anti-TIA-1. The results indicated that out of 37 cases 4 were NLPHL, 33 were CHL including 6 of MCHL, 14 of NSHL, 13 of LRHL. In addition, 10 cases (1 was NLPHL, 4 were NSHL, 5 were LRHL) were involved in the analysis of T/B ratios. The ratio of T/B in NLPHL was 0.28 +/- 0.07, in CHL 4.34 +/- 2.45 (p = 0.001), in CHL the ratio was LRHL > NSHL > MCHL (p = 0.649); CD4(+)/CD8(+) ratio in NLPHL was 4.55 +/- 1.28, in CHL 4.10 +/- 1.50 (p = 0.574), in CHL it was MCHL > NSHL > LRHL (p = 0.037); GrB(+)/TIA-1(+) ratio in NLPHL was 0.71 +/- 0.57, in CHL 0.74 +/- 0.39, it was MCHL > NSHL > LRHL (p > 0.05). It is conduced that immune cell composition of the diagnostic HL lymph node represents the immune microenvironment. It is different between NLPHL and CHL in terms of the T- and B-lymphocyte distribution. NLPHL is of unique feature. The subtypes of CHL are of peculiar. The T/B ratio is in order as LRHL > NSHL > MCHL, but CD4(+)/CD8(+) and GrB(+)/TIA-1(+) ratios are in the opposite order. Combining with prognosis of subtypes of CHL i.e, LRHL > NSHL > MCHL, these data suggest that low ratio of T/B with high ratios of CD4(+)/CD8(+) and GrB(+)/TIA-1(+) may represent biological markers predicting an unfavorable outcome of CHL subtypes.
Subject(s)
Adolescent , Adult , Aged , Child , Child, Preschool , Female , Humans , Male , Middle Aged , Young Adult , B-Lymphocytes , Cell Biology , CD4-CD8 Ratio , Hodgkin Disease , Allergy and Immunology , Pathology , Immunophenotyping , Lymphocytes , Allergy and Immunology , Pathology , T-Lymphocytes , Cell BiologyABSTRACT
<p><b>OBJECTIVE</b>To observe the changes in the activity of dendritic cells (DCs) after carcino-embryonic antigen (CEA) gene transfection mediated by recombinant adeno-associated virus type2 (rAAV) and tumor cell lysate.</p><p><b>METHODS</b>Immature DCs isolated from peripheral blood monocytes of HLA-A11-positive healthy volunteers were infected with the rAAV carrying CEA gene or loaded with tumor cell lysate. The surface markers of the DCs such as CD40, CD 1alpha, and CD86 were analyzed by flow cytometry. Interleukin-12 (IL-12) in the supernatants of DCs and interferon-gamma (IFN-gamma) released by the cytotoxic T lymphocytes (CTLs) were determined by ELISA detection kit. The specific killing activity of CTL against LoVo cells was assessed by MTT assay.</p><p><b>RESULTS</b>The DCs following antigen loading with the two methods both highly expressed CD40, CD86 and IL-12, and induced specific CTL that specifically recognized and killed LoVo cells, but the killing effect resulting from rAAV infection of the DCs was much better than that induced by tumor cell lysate loading.</p><p><b>CONCLUSION</b>Both methods of antigen loading can induce mature DCs from peripheral blood monocyte cells, but rAAV infection of the DCs can be more effective than tumor cells lysate loading. DCs infected with rAAV may have the potential to serve as an adjuvant immunotherapy for patients with colorectal carcinoma.</p>
Subject(s)
Humans , B7-2 Antigen , Metabolism , CD40 Antigens , Metabolism , Cancer Vaccines , Allergy and Immunology , Carcinoembryonic Antigen , Genetics , Cell Line, Tumor , Colorectal Neoplasms , Therapeutics , Dendritic Cells , Allergy and Immunology , Metabolism , Dependovirus , Genetics , Genetic Vectors , Interleukin-12 , Metabolism , TransfectionABSTRACT
<p><b>OBJECTIVE</b>To investigate the effects of procyanidins on proliferation and apoptosis of a hormone-dependent prostate cancer cell line LNCaP in vitro.</p><p><b>METHODS</b>LNCaP cells were cultured in the presence of procyanidin (at 100, 200 and 300 microg/ml, respectively). After 24, 48 and 72 h of culture, the morphological changes of LNCaP cells were examined, and the inhibition of cell proliferation was measured by MTT assay and cell apoptosis evaluated by flow cytometry.</p><p><b>RESULTS</b>After procyanidins treatment, some LNCaP cells presented characteristic morphological changes. Procyanidins inhibited the growth of LNCaP cells in a concentration- and time-dependent manner. Flow cytometry analysis showed that procyanidins induced apoptosis of LNCaP cells and the percentage of apoptotic cells increased with the concentration of procyanidins administered and also the elongation of treatment time.</p><p><b>CONCLUSION</b>Procyanidins inhibit the proliferation and promote the apoptosis of prostate cancer cells.</p>