RESUMO
AIMS: We discovered that some adults with coronary heart disease (CHD) have a high density lipoprotein (HDL) subclass which induces human aortic smooth muscle cell (ASMC) apoptosis in vitro. The purpose of this investigation was to determine what properties differentiate apoptotic and non-apoptotic HDL subclasses in adults with and without CHD. METHODS AND RESULTS: Density gradient ultracentrifugation was used to measure the particle density distribution and to isolate two HDL subclass fractions, HDL2 and HDL3, from 21 individuals, including 12 without CHD. The HDL fractions were incubated with ASMCs for 24 h; apoptosis was quantitated relative to C2-ceramide and tumour necrosis factor-alpha (TNF-α). The observed effect of some HDL subclasses on apoptosis was â¼6-fold greater than TNF-α and â¼16-fold greater than the cell medium. We observed that apoptotic HDL was (i) predominately associated with the HDL2 subclass; (ii) almost exclusively found in individuals with a higher apoC-I serum level and a novel, higher molecular weight isoform of apoC-I; and (iii) more common in adults with CHD, the majority of whom had high (>60 mg/dL) HDL-C levels. CONCLUSIONS: Some HDL subclasses enriched in a novel isoform of apoC-I induce extensive ASMC apoptosis in vitro. Individuals with this apoptotic HDL phenotype generally have higher apoC-I and HDL-C levels consistent with an inhibitory effect of apoC-I on cholesteryl ester transfer protein activity. The association of this phenotype with processes that can promote plaque rupture may explain a source of CHD risk not accounted for by the classical risk factors.
Assuntos
Apolipoproteína C-I/fisiologia , Apoptose , Lipoproteínas HDL/fisiologia , Miócitos de Músculo Liso/fisiologia , Adulto , Idoso , Idoso de 80 Anos ou mais , Apolipoproteína C-I/análise , Proteínas de Transferência de Ésteres de Colesterol/análise , Feminino , Humanos , Lipoproteínas HDL/análise , Lipoproteínas HDL/classificação , Masculino , Pessoa de Meia-Idade , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por MatrizRESUMO
Diagnosis of malignant pleural mesothelioma (MM) is limited. Novel proteomic techno_logies can be utilized to discover changes in expression of pleural proteins that might have diagnostic value. The objective of this study was to detect protein profiles that could be used to identify malignant pleural mesothelioma with surface enhanced laser desorption/ionization time-of-flight (SELDI-TOF) mass spectrometry (MS). Pleural effusions were collected from patients with confirmed mesothelioma (n = 41) and from patients with effusions due to other causes ([n = 48] cancerous and non-cancerous). Samples were fractionated using anion exchange chromatography and bound to different types of ProteinChip array surfaces. All samples were also subjected to other commercially available immunoassays (human epididymes protein 4 [HE4], osteopontin [OPN], soluble mesothelin-related proteins [SMRP], and the cytokeratin 19 fragment [CYFRA 21-1]). Peak intensity data obtained by SELDI-TOF were subjected to classification algorithms in order to identify potential classifier peaks. A protein peak at m/z 6614 was characterized as apolipoprotein (Apo) CI. In this setting, the sensitivity and specificity of this potential biomarker was 76 % and 69 %, respectively. The area under the receiver operating characteristic curve (AUC) for Apo CI was 0.755, thereby outperforming OPN, HE4, and CYFRA 21-1. SMRP performed best with an AUC of 0.860 with a sensitivity of 83% and specificity of 74%. Our study validates the use of SMRP as a diagnostic marker for pleural mesothelioma and furthermore suggests that Apo CI levels could be used in the future to discriminate pleural mesothelioma from other causes of exudates.
Assuntos
Apolipoproteína C-I/análise , Biomarcadores Tumorais/análise , Mesotelioma/diagnóstico , Proteínas de Neoplasias/análise , Derrame Pleural Maligno/diagnóstico , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz , Adulto , Idoso , Idoso de 80 Anos ou mais , Área Sob a Curva , Feminino , Humanos , Imunoensaio , Masculino , Mesotelioma/química , Pessoa de Meia-Idade , Estadiamento de Neoplasias , Derrame Pleural Maligno/química , Prognóstico , Análise Serial de Proteínas , Proteoma/análise , Sensibilidade e EspecificidadeRESUMO
Androgens play a central role in prostate cancer pathogenesis, and hence most of the patients respond to androgen deprivation therapies. However, patients tend to relapse with aggressive prostate cancer, which has been termed as hormone refractory. To identify the proteins that mediate progression to the hormone-refractory state, we used protein-chip technology for mass profiling of patients' sera. This study included 16 patients with metastatic hormone-refractory prostate cancer who were initially treated with androgen deprivation therapy. Serum samples were collected from each patient at five time points: point A, pre-treatment; point B, at the nadir of the prostate-specific antigen (PSA) level; point C, PSA failure; point D, the early hormone-refractory phase; and point E, the late hormone-refractory phase. Using surface-enhanced laser desorption/ionization time-of-flight mass spectrometry, we performed protein mass profiling of the patients' sera and identified a 6 640-Da peak that increased with disease progression. Target proteins were partially purified, and by amino acid sequencing the peak was identified as a fragment of apolipoprotein C-I (ApoC-I). Serum ApoC-I protein levels increased with disease progression. On immunohistochemical analysis, the ApoC-I protein was found localized to the cytoplasm of the hormone-refractory cancer cells. In this study, we showed an increase in serum ApoC-I protein levels in prostate cancer patients during their progression to the hormone-refractory state, which suggests that ApoC-I protein is related to progression of prostate cancer. However, as the exact role of ApoC-I in prostate cancer pathogenesis is unclear, further research is required.
Assuntos
Antineoplásicos Hormonais/uso terapêutico , Apolipoproteína C-I , Resistencia a Medicamentos Antineoplásicos , Neoplasias da Próstata/tratamento farmacológico , Neoplasias da Próstata/metabolismo , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz , Idoso , Sequência de Aminoácidos , Apolipoproteína C-I/análise , Apolipoproteína C-I/sangue , Apolipoproteína C-I/isolamento & purificação , Western Blotting , Linhagem Celular , Progressão da Doença , Humanos , Imuno-Histoquímica , Masculino , Pessoa de Meia-Idade , Dados de Sequência Molecular , Prognóstico , Análise Serial de ProteínasRESUMO
Previous studies have shown that postprandial triglyceride-rich lipoproteins (TRLs) are enriched with apolipoprotein-C-I (apoC-I) in healthy individuals with increased intima-media thickness and in patients with coronary artery disease. The purpose of the present study was to determine apoC-I in TRL in persons with carotid atherosclerosis and its relation to plaque area. A population-based case (n = 42)-control (n = 39) study was conducted in persons with carotid atherosclerosis, assessed by B-mode ultrasound, and healthy controls. VLDL (Sf 20-400) was isolated in the fasting state and 4 h after ingestion of a standard fat meal. In the fasting state, persons with carotid atherosclerosis had increased number of apoC-I per VLDL-particle compared to persons without carotid atherosclerosis (8.6 +/- 5.4 vs. 6.3 +/- 4.2, P = 0.018). Total plaque area increased linearly (P = 0.017) across tertiles of apoC-I per VLDL-particle. In the postprandial state, a similar increase in the number of apoC-I per VLDL-particle occurred in both cases and controls (P < 0.001), but no significant difference was observed between groups. The number of apoC-I per VLDL-particle in the fasting state was accompanied by delayed clearance of TRL in the postprandial state, and associated with cholesterol enrichment of the VLDL-particles. Our findings support the concept that the number of apoC-I per VLDL-particle may be of importance for initiation and progression of atherosclerosis.