Asunto(s)
Proteínas Adaptadoras Transductoras de Señales/inmunología , Autoanticuerpos/sangre , Biomarcadores de Tumor/sangre , Proteínas Inhibidoras de la Apoptosis/inmunología , Proteínas Asociadas a Microtúbulos/inmunología , Proteínas de Neoplasias/inmunología , Neoplasias/diagnóstico , Apoptosis/inmunología , Biomarcadores/sangre , Ensayo de Inmunoadsorción Enzimática/métodos , Valores de Referencia , Manejo de Especímenes , SurvivinRESUMEN
BACKGROUND: We previously reported that survivin-2B, a splicing variant of survivin, was expressed in various types of tumors and that survivin-2B peptide might serve as a potent immunogenic cancer vaccine. The objective of this study was to examine the toxicity of and to clinically and immunologically evaluate survivin-2B peptide in a phase I clinical study for patients with advanced or recurrent breast cancer. METHODS: We set up two protocols. In the first protocol, 10 patients were vaccinated with escalating doses (0.1-1.0 mg) of survivin-2B peptide alone 4 times every 2 weeks. In the second protocol, 4 patients were vaccinated with the peptide at a dose of 1.0 mg mixed with IFA 4 times every 2 weeks. RESULTS: In the first protocol, no adverse events were observed during or after vaccination. In the second protocol, two patients had induration at the injection site. One patient had general malaise (grade 1), and another had general malaise (grade 1) and fever (grade 1). Peptide vaccination was well tolerated in all patients. In the first protocol, tumor marker levels increased in 8 patients, slightly decreased in 1 patient and were within the normal range during this clinical trial in 1 patient. With regard to tumor size, two patients were considered to have stable disease (SD). Immunologically, in 3 of the 10 patients (30%), an increase of the peptide-specific CTL frequency was detected. In the second protocol, an increase of the peptide-specific CTL frequency was detected in all 4 patients (100%), although there were no significant beneficial clinical responses. ELISPOT assay showed peptide-specific IFN-gamma responses in 2 patients in whom the peptide-specific CTL frequency in tetramer staining also was increased in both protocols. CONCLUSION: This phase I clinical study revealed that survivin-2B peptide vaccination was well tolerated. The vaccination with survivin-2B peptide mixed with IFA increased the frequency of peptide-specific CTL more effectively than vaccination with the peptide alone, although neither vaccination could induce efficient clinical responses. Considering the above, the addition of another effectual adjuvant such as a cytokine, heat shock protein, etc. to the vaccination with survivin-2B peptide mixed with IFA might induce improved immunological and clinical responses.
Asunto(s)
Neoplasias de la Mama/inmunología , Neoplasias de la Mama/terapia , Vacunas contra el Cáncer/uso terapéutico , Carcinoma/inmunología , Carcinoma/terapia , Proteínas Asociadas a Microtúbulos/inmunología , Proteínas de Neoplasias/inmunología , Adulto , Anciano , Formación de Anticuerpos/fisiología , Neoplasias de la Mama/patología , Vacunas contra el Cáncer/inmunología , Carcinoma/patología , Femenino , Humanos , Inmunoterapia , Proteínas Inhibidoras de la Apoptosis/inmunología , Proteínas Asociadas a Microtúbulos/química , Persona de Mediana Edad , Proteínas de Neoplasias/química , Fragmentos de Péptidos/inmunología , Recurrencia , Pruebas Serológicas , SurvivinRESUMEN
We previously reported that beta-SQAG9 liposome, a sulfonoglycolipid extracted from sea urchin intestines, had a protective effect against hepatic ischemia reperfusion (I/R) injury. In this study, we made a detailed investigation of this protective effect and its mechanism. Rats were pretreated either with beta-SQAG9 liposome (treated group) or with phosphate-buffered saline solution (control group). Thereafter, they were subjected to partial hepatic I/R. The serum levels of aspartate aminotransferase, alanine aminotransferase, and lactate dehydrogenase were measured, and histological damage was evaluated with hematoxylin and eosin staining. To investigate the protective mechanism of beta-SQAG9 liposome on I/R injury, the serum levels and the tissue messenger RNA levels of TNF-alpha and IL-1beta were measured, and polymorphonuclear neutrophil (PMN) infiltration was histologically evaluated by immunohistochemistry. Moreover, to investigate an interaction between beta-SQAG9 liposome and L-selectin on PMNs, flow cytometric analysis and immunofluorescence were performed. beta-SQAG9 liposome reduced the hepatic I/R injury. The pretreatment with beta-SQAG9 liposome reduced the PMN infiltration into the liver parenchyma. On the other hand, there was no apparent difference in the serum levels and the tissue messenger RNA levels of the proinflammatory cytokines between the two groups. Thus, beta-SQAG9 liposome might reduce the hepatic I/R injury by inhibition of the PMN infiltration into the liver parenchyma, which was independent of the regulation of cytokine production. Moreover, we demonstrated that beta-SQAG9 liposome specifically bound to L-selectin on PMN cell surface, which mediated the PMN infiltration. beta-SQAG9 liposome might competitively antagonize L-selectin on PMNs and suppress the subsequent PMN infiltration, resulting in the reduction in I/R injury.
Asunto(s)
Glucolípidos/farmacología , Hígado/irrigación sanguínea , Hígado/lesiones , Daño por Reperfusión/prevención & control , Alanina Transaminasa/sangre , Animales , Aspartato Aminotransferasas/sangre , Glucolípidos/administración & dosificación , Glucolípidos/aislamiento & purificación , Glucolípidos/metabolismo , Técnicas In Vitro , Interleucina-1beta/sangre , Interleucina-1beta/genética , L-Lactato Deshidrogenasa/sangre , Selectina L/metabolismo , Liposomas , Hígado/efectos de los fármacos , Hígado/metabolismo , Masculino , Neutrófilos/metabolismo , ARN Mensajero/genética , ARN Mensajero/metabolismo , Ratas , Ratas Endogámicas Lew , Daño por Reperfusión/genética , Daño por Reperfusión/metabolismo , Daño por Reperfusión/patología , Erizos de Mar/química , Factor de Necrosis Tumoral alfa/sangre , Factor de Necrosis Tumoral alfa/genéticaRESUMEN
BACKGROUND: Although prewarming (PW) can reduce the confounding agglutination of cold-reactive antibodies when testing for warm-reactive antibodies, PW also can adversely affect the detection of warm-reactive antibodies. This study was conducted to determine PW conditions that optimized Rh antibody detection. STUDY DESIGN AND METHODS: In 38 plasma samples with Rh system antibodies detected by any of four methods, and in 8 other samples with cold-reactive antibodies, reactivity was assessed by titer and score. RESULTS: PW to 37 degrees C often reduced reaction scores by all methods especially the low-ionic-strength saline indirect antiglobulin test and bromelin methods. In analyses warming red blood cell (RBC) suspensions or plasma, the reaction scores were decreased only when the RBC suspension was warmed, suggesting that RBC PW decreased Rh antibody reactivity. The warming period ranging from 5 to 30 minutes all decreased Rh system reaction scores, an effect persisting up to 120 minutes. At lower temperatures (27, 30, and 33 degrees C), numbers of samples showing decreased reaction score for Rh system antibodies decreased in a temperature-dependent manner. Testing at 27 degrees C also permitted some agglutination involving cold-reactive antibodies, whereas higher temperatures successfully suppressed their agglutination. CONCLUSION: PW at 30 degrees C minimizes problems in accurately detecting Rh system antibodies.
Asunto(s)
Calor , Técnicas Inmunológicas , Isoanticuerpos/inmunología , Aglutinación , Unión Competitiva , Bromelaínas/química , Crioglobulinas/inmunología , Eritrocitos/inmunología , Humanos , Concentración Osmolar , Polietilenglicoles/química , Factores de TiempoRESUMEN
BACKGROUND: The anti-apoptotic molecule survivin is expressed in human cancers of various origins. Since this molecule possesses multiple functions, including apoptosis inhibition, cell cycle promotion and enhancement of Fas ligand expression, survivin has attracted growing attention as a target in cancer treatment. A survivin-specific small inhibitory RNA (siRNA) was introduced into pancreatic cancer cells to investigate its effect on cancer cell growth. MATERIALS AND METHODS: Survivin mRNA and protein expression were examined by RT-PCR and Western blotting, respectively. DNA histogram analysis was performed using a flow cytometer. RESULTS: The introduction of survivin-specific siRNA reduced survivin mRNA and protein expression in PANC-1 cells by over 90% and to an undetected amount, respectively, and induced growth inhibition. The siRNA transfectants showed pronounced morphological changes including enlargement of cells and multinucleation. siRNA transfectants did not show cell cycle arrest, but underwent apoptosis. CONCLUSION: Our data suggest that the use of survivin-specific siRNA deserves further investigation as a novel approach to cancer therapy.
Asunto(s)
Proteínas Asociadas a Microtúbulos/genética , Proteínas de Neoplasias/genética , Neoplasias Pancreáticas/genética , Neoplasias Pancreáticas/patología , ARN Interferente Pequeño/genética , Procesos de Crecimiento Celular/genética , Línea Celular Tumoral , Terapia Genética/métodos , Humanos , Proteínas Inhibidoras de la Apoptosis , Neoplasias Pancreáticas/terapia , Interferencia de ARN , Survivin , TransfecciónRESUMEN
OBJECTIVES: Autoantibodies to tumor-associated antigens including survivin have been detected in sera from patients with hepatocellular carcinoma (HCC). However, little is known about autoantibody responses to tumor-associated antigens in patients with chronic hepatitis, which strongly predisposes to development of HCC. METHODS: We subjected sera from 57 patients with chronic hepatitis and 29 patients with HCC to an enzyme-linked immunosorbent assay (ELISA) using a full-length recombinant survivin protein. A cutoff value for positivity was determined as the mean absorbance +2SD for sera from healthy volunteers. RESULTS: In patients with chronic viral hepatitis, elevated anti-survivin antibodies were detected in 10 of 57 sera (17.5%); in HCC patients, such elevation were detected in 7 of 29 sera (24.1%). The levels of anti-survivin antibodies in HCC patients with HCV infection were significantly higher than those in the healthy control and HCC patients with HBV infection. However, there were no significant differences in the levels of anti-survivin antibodies between HCV and HCC patients with HCV infection. CONCLUSIONS: We demonstrated that elevated anti-survivin antibodies were detected for the first time in patients with chronic viral hepatitis. The results suggest that the levels of anti-survivin antibodies have no association with the progression of HCV or HBV to HCC.
Asunto(s)
Autoanticuerpos/biosíntesis , Carcinoma Hepatocelular/inmunología , Hepatitis B Crónica/inmunología , Hepatitis C Crónica/inmunología , Neoplasias Hepáticas/inmunología , Proteínas Asociadas a Microtúbulos/inmunología , Proteínas de Neoplasias/inmunología , Autoanticuerpos/sangre , Ensayo de Inmunoadsorción Enzimática , Virus de la Hepatitis B/inmunología , Humanos , Proteínas Inhibidoras de la Apoptosis , SurvivinAsunto(s)
Biomarcadores de Tumor/análisis , Neoplasias de la Mama/diagnóstico , Antígeno Carcinoembrionario/análisis , Pezones/metabolismo , Enfermedades de la Mama/diagnóstico , Femenino , Humanos , Técnicas para Inmunoenzimas/métodos , Juego de Reactivos para Diagnóstico , Valores de Referencia , Manejo de EspecímenesAsunto(s)
Proteínas Adaptadoras Transductoras de Señales/inmunología , Proteínas Asociadas a Microtúbulos/inmunología , Proteínas de Neoplasias/inmunología , Neoplasias/diagnóstico , Apoptosis/inmunología , Autoanticuerpos/sangre , Biomarcadores/sangre , Ensayo de Inmunoadsorción Enzimática/métodos , Humanos , Proteínas Inhibidoras de la Apoptosis , SurvivinRESUMEN
BACKGROUND: Survivin and livin are highly expressed in cancer cells and transformed cells, but show little or no expression in normal differentiated tissues. Human antibody responses to tumor-associated antigens have been detected, but little is known about the response to survivin and livin in breast cancer patients. METHODS: We examined the prevalence of anti-survivin and livin antibodies in breast cancer patients with a specific enzyme-linked immunosorbent assay (ELISA) using recombinant protein. RESULTS: Using a cutoff value for positivity determined as the mean absorbance +2SD for healthy control samples, sera from 11 of 46 breast cancer patients (23.9%) were positive by the ELISA using recombinant survivin protein. Of 46 samples from the same breast cancer patients, 15 (32.6%) were positive for anti-livin antibodies. In addition, 24 (52.2%) were positive for 1 or both ELISAs using the respective proteins. Intensity of anti-livin antibody responses did not correlate with intensity of anti-survivin responses. CONCLUSIONS: Anti-livin antibodies were detected in sera from breast cancer patients by an anti-livin ELISA using full-length recombinant livin protein. Like survivin, livin may act as a major cancer antigen in breast cancer patients.
Asunto(s)
Proteínas Adaptadoras Transductoras de Señales/sangre , Proteínas Adaptadoras Transductoras de Señales/inmunología , Autoanticuerpos/inmunología , Neoplasias de la Mama/sangre , Neoplasias de la Mama/inmunología , Proteínas Inhibidoras de la Apoptosis/sangre , Proteínas Inhibidoras de la Apoptosis/inmunología , Proteínas Asociadas a Microtúbulos/sangre , Proteínas Asociadas a Microtúbulos/inmunología , Proteínas de Neoplasias/sangre , Proteínas de Neoplasias/inmunología , Absorción , Autoanticuerpos/sangre , Biomarcadores de Tumor/inmunología , Humanos , SurvivinRESUMEN
Beclin 1, identified as a Bcl-2-interacting protein, is known to enhance autophagy. However, the effect of Beclin 1 on apoptotic signaling has remained unclear. Here, we show that overexpression of Beclin 1 in MKN28 human gastric cancer cells augmented cis-diamminedichloroplatinum (CDDP)-induced apoptosis. Conversely, "knockdown" of Beclin 1 by a small inhibitory RNA in MKN 1 cells attenuated this cytotoxicity. Furthermore, not only caspase-3/7 activities, but also caspase-9 activity was increased in Beclin 1 gene transfectants treated with CDDP, and caspase-9 inhibitor completely abolished augmentation of CDDP-induced apoptosis by Beclin 1 as did a caspase-3 inhibitor. Thus, Beclin 1 augments CDDP-induced apoptosis through enhancing caspase-9 activity and functions as a pro-apoptotic molecule.
Asunto(s)
Antineoplásicos/toxicidad , Apoptosis/efectos de los fármacos , Caspasas/metabolismo , Cisplatino/toxicidad , Proteínas/metabolismo , Proteínas Reguladoras de la Apoptosis , Beclina-1 , Western Blotting , Caspasa 3 , Caspasa 7 , Caspasa 8 , Caspasa 9 , Línea Celular Tumoral , Activación Enzimática/efectos de los fármacos , Precursores Enzimáticos/metabolismo , Regulación Neoplásica de la Expresión Génica , Genes Supresores de Tumor , Humanos , Proteínas de la Membrana , Poli(ADP-Ribosa) Polimerasas/metabolismo , Pruebas de Precipitina , ARN Mensajero/metabolismo , ARN Interferente Pequeño/metabolismo , Neoplasias Gástricas/genética , Neoplasias Gástricas/patologíaRESUMEN
Survivin and livin are highly expressed in cancer cells and transformed cells, but show little or no expression in normal differentiated tissues. Although human antibody responses to cancer-associated antigens have been detected, the response to livin has not yet been described in lung cancer patients. We examined prevalence of anti-livin antibodies in such patients with a specific enzyme-linked immunosorbent assay (ELISA) using recombinant protein. Using a cutoff value for positivity determined as the mean absorbance +2S.D. for healthy control samples, 19 of 37 lung cancer patients (51.3%) were positive for anti-livin antibodies. Of 31 samples from the same lung cancer patients, 18 (58.1%) were positive for anti-survivin antibodies. When sera from 31 lung cancer patients were assessed simultaneously by anti-survivin and anti-livin ELISAs. Twenty-one patients (71%) were positive for survivin, livin, or both. Intensity of anti-livin antibody responses did not correlate with intensity of anti-survivin responses. Like anti-survivin antibodies, anti-livin antibodies, thus, can be detected in many lung cancer patients. Testing for both antibodies together may prove useful in detecting lung cancer, but more extensive studies are needed to establish the clinical significance of anti-livin antibodies.
Asunto(s)
Proteínas Adaptadoras Transductoras de Señales/inmunología , Autoanticuerpos/análisis , Carcinoma de Pulmón de Células no Pequeñas/inmunología , Carcinoma de Células Pequeñas/inmunología , Neoplasias Pulmonares/inmunología , Proteínas Asociadas a Microtúbulos/inmunología , Proteínas de Neoplasias/inmunología , Anticuerpos Antineoplásicos , Formación de Anticuerpos , Estudios de Casos y Controles , Ensayo de Inmunoadsorción Enzimática , Humanos , Proteínas Inhibidoras de la Apoptosis , Neoplasias Pulmonares/fisiopatología , SurvivinRESUMEN
Suppression of apoptosis is thought to contribute to carcinogenesis. Survivin, a member of the inhibitor-of-apoptosis family, blocks apoptotic signaling activated by various cellular stresses. Since elevated expression of survivin observed in human cancers of varied origin was associated with poor patient survival, survivin has attracted growing attention as a potential target for cancer treatment. Immortalization of cells also is required for carcinogenesis; telomere length maintenance by telomerase is required for cancer cells to proliferate indefinitely. Yet how cancer cells activate telomerase remains unclear. We therefore examined possible interrelationships between survivin expression and telomerase activity. Correlation between survivin and human telomerase reverse transcriptase (hTERT) expression was observed in colon cancer tissues, and overexpression of survivin enhanced telomerase activity by up-regulation of hTERT expression in LS180 human colon cancer cells. DNA-binding activities of specificity protein 1 (Sp1) and c-Myc to the hTERT core promoter were increased in survivin gene transfectant cells. Phosphorylation of Sp1 and c-Myc at serine and threonine residues was enhanced by survivin, while total amounts of these proteins were unchanged. Further, "knockdown" of survivin by a small inhibitory RNA decreased Sp1 and c-Myc phosphorylation. Thus survivin participates not only in inhibition of apoptosis, but also in prolonging cellular lifespan.
Asunto(s)
Neoplasias del Colon/enzimología , Proteínas Asociadas a Microtúbulos/fisiología , Proteínas Proto-Oncogénicas c-myc/metabolismo , Factor de Transcripción Sp1/metabolismo , Telomerasa/metabolismo , Regulación hacia Arriba , Cromonas/farmacología , Neoplasias del Colon/genética , Neoplasias del Colon/metabolismo , Proteínas de Unión al ADN , Regulación Neoplásica de la Expresión Génica , Humanos , Proteínas Inhibidoras de la Apoptosis , Proteínas Asociadas a Microtúbulos/genética , Morfolinas/farmacología , Proteínas de Neoplasias , Inhibidores de las Quinasa Fosfoinosítidos-3 , Fosforilación , Regiones Promotoras Genéticas/genética , ARN Interferente Pequeño/genética , Survivin , Telomerasa/genética , Transcripción GenéticaRESUMEN
It has been suggested that suppression of apoptosis may contribute to the development and progression of cancer. Anti-apoptotic survivin and livin genes are highly expressed in cancer cells and transformed cells, but show little or no expression in normal differentiated tissues. However, there are no available data concerning livin expression in non-small-cell lung cancer (NSCLC). We therefore measured livin mRNA and survivin mRNA expression in 38 NSCLC cancer samples and 15 paired non-cancerous lung tissue samples using a quantitative reverse transcription-polymerase chain reaction (RT-PCR). While both mRNAs showed higher expression in cancers than non-cancerous tissues (P < 0.001), livin mRNA and survivin mRNA expression did not correlate with one another. When a cut-off value for positivity was set at the mean + S.D. for expression values in non-cancerous tissues, positivity rates for livin mRNA and survivin mRNA expression were 76.3% (29 of 38) and 36.8% (14 of 38) in lung cancers and 6.7% (1 of 15) and 0% (0 of 15), respectively, in paired non-cancerous lung tissue samples. Livin mRNA and survivin mRNA expression in tumors were up-regulated in 23 of 31 (74.2%) early-stage NSCLC patients and 11 of 31 (35.5%), respectively. Expression of both mRNAs in tumors varied independently of tumor histology. These results support the possibility that the livin gene may play a role in NSCLC development and increased expression of livin mRNA may serve as a new target for lung cancer treatment as well as survivin.
Asunto(s)
Proteínas Adaptadoras Transductoras de Señales/biosíntesis , Proteínas Adaptadoras Transductoras de Señales/genética , Apoptosis , Carcinoma de Pulmón de Células no Pequeñas/genética , Carcinoma de Pulmón de Células no Pequeñas/fisiopatología , Perfilación de la Expresión Génica , Neoplasias Pulmonares/genética , Neoplasias Pulmonares/fisiopatología , Proteínas Asociadas a Microtúbulos/biosíntesis , Proteínas Asociadas a Microtúbulos/genética , Proteínas de Neoplasias/biosíntesis , Proteínas de Neoplasias/genética , Estudios de Casos y Controles , Transformación Celular Neoplásica , Progresión de la Enfermedad , Humanos , Proteínas Inhibidoras de la Apoptosis , Estadificación de Neoplasias , ARN Mensajero/análisis , ARN Mensajero/biosíntesis , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Survivin , Dedos de ZincRESUMEN
Expression of survivin, a member of the inhibitor-of-apoptosis (IAP) family, is elevated in fetal tissues and in various human cancers originating in the breast, lung, prostate, colon, pancreas, and stomach. Since overexpression of the survivin gene has been linked to poor patient survival in several cancers, survivin may be an important prognostic marker. Mechanisms up-regulating survivin gene expression in cancer are poorly understood. Recently, wild-type p53 was found to repress expression of the survivin gene by binding to the survivin promoter, thereby inhibiting promoter activity. Further, loss of heterozygosity (LOH) at 17p13 distal to the p53 gene is associated with more aggressive behavior of breast cancers. We therefore tested the hypothesis that not only p53 gene mutation but also LOH at 17p13 can up-regulate survivin expression in breast cancer. Survivin mRNA expression was greater in cancers than in uninvolved tissues (p < 0.0001). Mutations of the p53 gene were detected in 5 of 25 tumors; higher survivin gene expression was evident in these. LOH at the D17S938 locus (17p13.1) was found in 10 of 25 tumors, and most of these also showed increased survivin gene expression. Thus expression of survivin may be regulated not only by p53 but additionally by a putative tumor suppressor gene located at 17p13 distal to the p53 gene.
Asunto(s)
Neoplasias de la Mama/genética , Cromosomas Humanos Par 17/genética , Genes p53 , Proteínas Asociadas a Microtúbulos/biosíntesis , Adulto , Anciano , Biomarcadores de Tumor/análisis , Neoplasias de la Mama/fisiopatología , Análisis Mutacional de ADN , Femenino , Perfilación de la Expresión Génica , Humanos , Proteínas Inhibidoras de la Apoptosis , Pérdida de Heterocigocidad , Persona de Mediana Edad , Proteínas de Neoplasias , Pronóstico , Regiones Promotoras Genéticas/genética , ARN Mensajero/biosíntesis , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Survivin , Regulación hacia ArribaRESUMEN
Survivin is a member of the inhibitor of apoptosis protein (IAP) family containing a single baculovirus IAP repeat domain. It is expressed during fetal development but becomes undetectable in terminally differentiated normal adult tissues. We previously reported that survivin and its splicing variant survivin-2B was expressed abundantly in various types of tumor tissues as well as tumor cell lines and was suitable as a target antigen for active-specific anti-cancer immunization. Subsequently, we identified an HLA-A24-restricted antigenic peptide, survivin-2B80-88 (AYACNTSTL) recognized by CD8+ cytotoxic T lymphocytes (CTLs). We, therefore, started a phase I clinical study assessing the efficacy of survivin-2B peptide vaccination in patients with advanced or recurrent colorectal cancer expressing survivin. Vaccinations with survivin-2B peptide were given subcutaneously six times at 14-day intervals. Of 15 patients who finished receiving the vaccination schedule, three suffered slight toxicities, including anemia (grade 2), general malaise (grade 1), and fever (grade 1). No severe adverse events were observed in any patient. In 6 patients, tumor marker levels (CEA and CA19-9) decreased transiently during the period of vaccination. Slight reduction of the tumor volume was observed in one patient, which was considered a minor responder. No changes were noted in three patients while the remaining eleven patients experienced tumor progression. Analysis of peripheral blood lymphocytes of one patient using HLA-A24/peptide tetramers revealed an increase in peptide-specific CTL frequency from 0.09% to 0.35% of CD8+ T cells after 4 vaccinations. This phase I clinical study indicates that survivin-2B peptide-based vaccination is safe and should be further considered for potential immune and clinical efficacy in HLA-A24-expression patients with colorectal cancer.
RESUMEN
Cancer cells are thought to possess mechanisms for evading the host's immune surveillance system. Survivin, a member of the inhibitor-of-apoptosis family overexpressed by cancer cells, inhibits Fas-mediated apoptosis induced by immune cells. In addition, cancer cells express Fas ligand (FasL) on their surfaces as a counterattack against immune cells. Mechanisms by which cancer cells express FasL, including involvement of survivin, are unclear. In the present study, we demonstrated that survivin up-regulated FasL expression and investigated how this might occur. Quantitative immunostaining showed correlation between survivin and FasL protein expression in colon cancer tissues (r=0.79). FasL expression was up-regulated in LS180 colon cancer cells transfected with the survivin gene. Transfectants showed increased cytotoxicity against a Fas-sensitive human T leukemia cell line, Jurkat. In contrast, FasL expression was down-regulated in SW480 cells transfected with a small inhibitory RNA to prevent survivin expression. Survivin gene transfectants showed increased DNA binding of transcription factor specificity protein 1 (Sp1) to the FasL promoter, and up-regulation of Sp1 phosphorylation at serine and threonine residues; the total amount of Sp1 was unchanged. Thus, survivin enables cancer cells not only to suppress immune cell attack by inhibiting Fas-mediated apoptotic signaling, but to attack immune cells by induction of FasL.
Asunto(s)
Adyuvantes Inmunológicos/fisiología , Neoplasias del Colon/genética , Neoplasias del Colon/inmunología , Glicoproteínas de Membrana/biosíntesis , Proteínas Asociadas a Microtúbulos/fisiología , Factor de Transcripción Sp1/fisiología , Regulación hacia Arriba/inmunología , Adyuvantes Inmunológicos/biosíntesis , Adyuvantes Inmunológicos/genética , Línea Celular Tumoral , Neoplasias del Colon/metabolismo , Regulación hacia Abajo/inmunología , Proteína Ligando Fas , Humanos , Proteínas Inhibidoras de la Apoptosis , Células Jurkat , Ligandos , Glicoproteínas de Membrana/antagonistas & inhibidores , Glicoproteínas de Membrana/genética , Glicoproteínas de Membrana/metabolismo , Proteínas Asociadas a Microtúbulos/antagonistas & inhibidores , Proteínas Asociadas a Microtúbulos/biosíntesis , Proteínas Asociadas a Microtúbulos/genética , Proteínas de Neoplasias , Fosforilación , Regiones Promotoras Genéticas/inmunología , Factor de Transcripción Sp1/metabolismo , Survivin , Transcripción Genética/inmunología , Transfección , Receptor fas/metabolismoRESUMEN
Survivin, a member of the inhibitor-of-apoptosis family, inhibits apoptosis by blocking caspase-3 and -7 activation. Gastric cancer, which is among the most intractable of malignant tumors, is known for resistance to various drugs, including cis-diamminedichloroplatinum (CDDP). Since this agent induces apoptosis via caspase-3 activation, survivin may mediate the drug resistance. We investigated survivin messenger RNA (mRNA) expression in gastric cancers and the relationship between expression and sensitivity to CDDP. Expression of the survivin gene was significantly up-regulated in gastric cancers compared to the tissues of normal mucosa, atrophic gastritis, and intestinal metaplasia (P < 0.0001) as assessed by a quantitative reverse transcription-polymerase chain reaction (RT-PCR), and was negatively associated with overall survival of patients who received CDDP-based chemotherapy. To investigate whether survivin is a resistance factor against CDDP-induced apoptosis, we transfected wild-type and dominant-negative mutants of the survivin gene into gastric cancer cells using a lipofection method. Overexpression of survivin protected MKN45 cells from CDDP-induced apoptosis. Expression of the dominant-negative mutant of the survivin gene sensitized NUGC-3 cells to drug-induced apoptosis. These results indicate that survivin may be pivotal in the development of gastric cancer and resistance to CDDP, and therefore controlling expression of the survivin gene may be therapeutically useful.
Asunto(s)
Antineoplásicos/farmacología , Cisplatino/farmacología , Resistencia a Antineoplásicos/fisiología , Proteínas Asociadas a Microtúbulos/biosíntesis , Neoplasias Gástricas/tratamiento farmacológico , Adulto , Anciano , Anciano de 80 o más Años , Apoptosis/efectos de los fármacos , Western Blotting , Línea Celular Tumoral , Femenino , Expresión Génica , Humanos , Proteínas Inhibidoras de la Apoptosis , Masculino , Proteínas Asociadas a Microtúbulos/genética , Persona de Mediana Edad , Proteínas de Neoplasias , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Neoplasias Gástricas/mortalidad , Survivin , Transfección , Regulación hacia ArribaRESUMEN
BACKGROUND AND AIMS: The recently identified aspartate protease gene ALP56 is up-regulated in human malignant tumors, including colorectal cancers, but the relationship remain unclear between ALP56 gene expression and clinicopathological findings, as well as when genetic alterations in ALP56 occur during the colorectal adenoma-carcinoma sequence. We therefore investigated expression of ALP56 mRNA in various human colorectal tissues. MATERIALS AND METHODS: We examined 18 colorectal adenomas 22 cancers, and 24 adjacent normal mucosal samples from patients undergoing conventional resection or endoscopic mucosal resection. Expression of ALP56 mRNA was determined by quantitative reverse-transcription polymerase chain reaction. RESULTS: Up-regulation of ALP56 gene transcription was observed in both adenomas and cancers compared to normal mucosa. ALP56 expression in exophytic adenomas was significantly greater than in flat adenomas. CONCLUSION: ALP56 may contribute to colorectal adenoma formation and to an exophytic growth pattern in these adenomas.