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1.
Lasers Med Sci ; 27(2): 445-52, 2012 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-21769639

RESUMO

Photodynamic therapy (PDT) is attracting attention because of its noticeable inhibitory effects on the growth of dermatological and other solid tumors. Here, we studied the use of PDT in systemic diseases such as leukemia, lymphoma, and metastatic cancer, for which tumor formation areas cannot be clearly compartmentalized. We developed a systemic PDT method and examined its effect in a leukemia mouse model. Growth inhibition of A20 cells (H-2(d), murine B-lymphoma/leukemia, and Balb/c origin) induced by PDT/Photodithazine was evaluated by EZ-Cytox assay. After PDT, changes in cell morphology were assessed by light microscopy. Induction of apoptosis, as well as changes in the cell cycle, were assessed by fluorescence-activated cell sorting (FACS) analysis. A20 cells were injected into Balb/c mice through the tail veins, and PDT was performed. A total of 10 mg kg(-1) body weight of Photodithazine concentration was injected intravenously. After 5 min, micro photofibers (diameter, 200 µm) were inserted into the tail veins and irradiated at 1,200 J with a laser. PDT inhibited growth of A20 cells and resulted in marked morphological changes. PDT also induced apoptosis and G1 arrest. In a leukemia mouse model, systemic PDT increased the survival rate (p < 0.01). This is the first report of the effects of systemic PDT in a leukemia animal model. PDT has been applied only locally in most cases, for example to solid tumors. This study provides experimental evidence that systemic PDT could effectively be applied to systemic and spread tumors, for which tumor formation areas cannot clearly be determined.


Assuntos
Apoptose/efeitos dos fármacos , Leucemia Experimental/tratamento farmacológico , Linfoma/tratamento farmacológico , Fotoquimioterapia/métodos , Fármacos Fotossensibilizantes/uso terapêutico , Animais , Linhagem Celular Tumoral , Citometria de Fluxo , Leucemia Experimental/patologia , Linfoma/patologia , Camundongos , Camundongos Endogâmicos BALB C , Transplante de Neoplasias
2.
Immunology ; 124(4): 461-8, 2008 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-18397271

RESUMO

Immunotherapy with photodynamic therapy (PDT) offers great promise as a new alternative for cancer treatment; however, its use remains experimental. Here we investigated the utility of adenoviral delivery of interleukin-12 (AdmIL-12) as an adjuvant for PDT in mouse tumour challenge model. PDT was performed by irradiating Radachlorin in C57BL/6 mice transplanted with TC-1 cells. PDT plus AdmIL-12 treatment for tumour suppression as well as specific immune responses were evaluated with the following tests: in vitro and in vivo tumour growth inhibition, interferon-gamma (IFN-gamma) and tumour necrosis factor-alpha (TNF-alpha) assay, and cytotoxic T lymphocyte (CTL) assay. Direct intratumoral injection of AdmIL-12 resulted in a significant suppression of tumour growth compared to the control group. Treatment of PDT along with AdmIL-12 further enhanced antitumour effects significantly higher than either AdmIL-12 or PDT alone. This combined treatment resulted in complete regression of 9-mm sized tumour in every animal. We also evaluated immune responses induced by these treatments. Combined treatment significantly increased the production level of IFN-gamma and TNF-alpha compared with that by AdmIL-12 or PDT alone. PDT plus AdmIL-12 enhanced antitumour immunity through increased expansion of the CTL subset mediated by CD8+ T cells. Taken together, these results indicate that the high anti-cancer activity of PDT with AdmIL-12 is a powerful tool against cancer therapy and is a promising subject for further investigation.


Assuntos
Terapia Genética/métodos , Papillomavirus Humano 16 , Interleucina-12/genética , Neoplasias Experimentais/terapia , Infecções por Papillomavirus/terapia , Fotoquimioterapia/métodos , Adenoviridae/genética , Animais , Linfócitos T CD8-Positivos/imunologia , Terapia Combinada , Citotoxicidade Imunológica , Feminino , Vetores Genéticos , Imunidade Celular , Interferon gama/biossíntese , Interleucina-12/imunologia , Camundongos , Camundongos Endogâmicos C57BL , Transplante de Neoplasias , Neoplasias Experimentais/imunologia , Neoplasias Experimentais/patologia , Infecções por Papillomavirus/imunologia , Infecções por Papillomavirus/patologia , Fármacos Fotossensibilizantes/farmacocinética , Linfócitos T Citotóxicos/imunologia , Fator de Necrose Tumoral alfa/biossíntese
3.
Arch Pharm Res ; 31(10): 1281-5, 2008 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-18958418

RESUMO

To explore the anticancer effects of the flavonoid quercetin on human breast cancer MDA-MB-453 cells via cell cycle regulation and the induction of apoptosis, the antiproliferative effect of quercetin was first examined by MTT assay. When MDA-MB-453 cells were treated with quercetin for various periods of time (3-24 hrs) and at various doses (1-100 microM), cell growth decreased significantly in a time-and dose-dependent manner. To elucidate the mechanism underlying the antiproliferative effect of quercetin, cell cycle progression and the induction of apoptosis in MDA-MB-453 cells exposed to 100 microM quercetin for 24 hrs were investigated. Quercetin caused a remarkable increase in the number of sub-G1 phase cells, and an Annexin-V assay revealed that exposure to quercetin affected apoptosis. Moreover, treatment with quercetin increased Bax expression but decreased Bcl-2 expression. Cleaved caspase-3 and PARP expression was also increased by quercetin. Thus, quercetin has probable anticancer activity. Our results suggest the existence of multiple pathways for the induction of cell cycle arrest and apoptosis by quercetin.


Assuntos
Antineoplásicos , Apoptose/efeitos dos fármacos , Neoplasias da Mama/tratamento farmacológico , Ciclo Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Quercetina/farmacologia , Western Blotting , Neoplasias da Mama/patologia , Linhagem Celular Tumoral , DNA de Neoplasias/biossíntese , DNA de Neoplasias/genética , Feminino , Citometria de Fluxo , Humanos , Proteínas de Neoplasias/biossíntese , Proteínas de Neoplasias/genética
4.
Int J Oncol ; 30(5): 1129-35, 2007 May.
Artigo em Inglês | MEDLINE | ID: mdl-17390014

RESUMO

As2O3 has been reported to induce apoptosis and inhibit the proliferation of various human cancer cells. We evaluated the ability of a novel arsenic compound, As4O6, along with As2O3 in vitro and in vivo. To examine the levels of apoptosis of HPV 16-positive SiHa cervical cancer cell, flow cytometry and Western blotting were employed at various time intervals after two arsenic compound treatments. Ingenuity Pathway Analysis (IPA) was applied to investigate the differential cell death pathway of As4O6 and As2O3. The results showed that As4O6 was more effective in suppressing SiHa cell growth in vitro and in vivo compared to As2O3. In addition, the cell cycle was arrested at the sub-G1 phase by As4O6. Western blot analysis showed that the proliferating cell nuclear antigen (PCNA) and Bcl-XL with sequence homology to Bcl-2 were significantly suppressed by As4O6. However, the apoptosis-related proteins such as p21 and Bax were overexpressed by As4O6. IPA suggested that there is a significant difference between As2O3- and As4O6-induced cell death pathways. Taken together, As4O6 has a specific cell death pathway and possesses more potent anti-tumor effects on human cervical cancer cells in vitro and in vivo.


Assuntos
Antineoplásicos/farmacologia , Apoptose , Arsenicais/farmacologia , Óxidos/farmacologia , Trióxido de Arsênio , Ciclo Celular , Linhagem Celular Tumoral , Proliferação de Células , Separação Celular , Ensaios de Seleção de Medicamentos Antitumorais , Citometria de Fluxo , Humanos , Neoplasias/tratamento farmacológico , Neoplasias/metabolismo , Antígeno Nuclear de Célula em Proliferação/química , Proteínas Proto-Oncogênicas c-bcl-2/química
5.
Hum Gene Ther ; 17(3): 347-52, 2006 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-16544983

RESUMO

Photodynamic therapy (PDT) has been reported to be effective for treating various tumors and to induce apoptosis in many tumor cells. In this study, we evaluated the ability of PDT combined with a tumor suppressor factor, recombinant adenovirus p53 (AdCMVp53), to induce apoptosis as well as cell growth inhibition in CaSki human cervical cancer cells and in nude mice with implanted CaSki cells. To examine levels of apoptosis, CaSki cells were treated with PDT and/or AdCMVp53, and an annexin V-staining assay was then conducted. In addition, Western blot analysis was done to identify p53 induction at the cellular and tumor tissue levels. PDT+AdCMVp53 cotreatment caused remarkable inhibition of CaSki cell proliferation, as compared with the individual treatments. In parallel with the inhibition of cell proliferation, the cotreatment caused a significantly greater increase in the annexin V-stained cell population compared with the individual treatments, as determined by fluorescence-activated cell-sorting analysis. The Western blotting assay also showed significantly more cellular p53 expressed after PDT+AdCMVp53 cotreatment than after each separate treatment. This was consistent with observations of tumor tissue in the mouse system. However, apoptosis- related protein, p21, was significantly suppressed by PDT+AdCMVp53 cotreatment, contrary to treatment with AdCMVp53 alone. Taken together, these findings suggest that PDT plus AdCMVp53 gene therapy exerts more potent antitumor effects on human cervical cancer cells, with induction of apoptosis at least through activation in p53 protein at the cellular and tumor tissue levels.


Assuntos
Adenoviridae/genética , Terapia Genética/métodos , Fotoquimioterapia , Fármacos Fotossensibilizantes/uso terapêutico , Proteína Supressora de Tumor p53/genética , Neoplasias do Colo do Útero/terapia , Animais , Apoptose/fisiologia , Apoptose/efeitos da radiação , Western Blotting , Linhagem Celular Tumoral , Proliferação de Células/efeitos da radiação , Terapia Combinada , Feminino , Vetores Genéticos/genética , Humanos , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Nus , Transfecção , Neoplasias do Colo do Útero/genética , Neoplasias do Colo do Útero/patologia , Ensaios Antitumorais Modelo de Xenoenxerto
6.
Biosens Bioelectron ; 20(11): 2236-43, 2005 May 15.
Artigo em Inglês | MEDLINE | ID: mdl-15797321

RESUMO

We investigated the time-course tumor growth suppression effects of recombinant adenovirus expressing p53 on human cervical cancer cells and cell-specific E7 protein-protein interactions in cell lysates using surface plasmon resonance (SPR) biosensor. Six HPV-infected human cervical cancer cell lines (HPV 16-positive cells, CaSki and SiHa cells; HPV 18-positive cells, HeLa and HeLaS3 cells; and HPV negative C33A and HT3 cells) were used. After infection with AdCMVp53, the cell-specific growth inhibition was studied in vitro and in vivo. Also, we produced the recombinant E7 oncoprotein of HPV 16 type and tested chip-based protein-protein interactions with each cell lysate. For each cervical cancer cell, differential cell growth inhibitions were shown via cell count assay and MTT assay. Note that the same trend in suppression levels was shown in CaSki, HeLa and in SiHa, HeLaS3, respectively. In contrast, infection with AdCMVLacZ showed increased cell growth in a manner similar to the negative control group. The levels of p53 protein were notably expressed in CaSki and HeLa more than in SiHa and HeLaS3 for 4 days. In contrast, p53 expression was continually maintained in C33A and HT3 for 6 days. After transfection AdCMVp53 into CaSki- and SiHa-xenografted nude mice, the size of tumor was remarkably decreased in SiHa cells as compared to AdCMVLacZ transfection. The SPR sensor surface was successfully modified with the recombinant E7 oncoprotein and showed cell-specific interactions between E7 and its target proteins from cell lysates. The anti-tumor effects were accomplished via differential role of p53-specific apoptotic cell death, which is dependent upon the cervical cancer cell line. Also, a molecular level understanding of cell-dependent protein interaction effects of recombinant E7 was shown.


Assuntos
Adenoviridae/genética , Técnicas Biossensoriais/métodos , Proteínas Oncogênicas Virais/metabolismo , Mapeamento de Interação de Proteínas/métodos , Ressonância de Plasmônio de Superfície/métodos , Proteína Supressora de Tumor p53/metabolismo , Neoplasias do Colo do Útero/metabolismo , Neoplasias do Colo do Útero/virologia , Técnicas Biossensoriais/instrumentação , Linhagem Celular Tumoral , Proliferação de Células , Feminino , Terapia Genética/métodos , Humanos , Proteínas E7 de Papillomavirus , Mapeamento de Interação de Proteínas/instrumentação , Ressonância de Plasmônio de Superfície/instrumentação , Transfecção/métodos , Proteína Supressora de Tumor p53/administração & dosagem , Proteína Supressora de Tumor p53/genética , Neoplasias do Colo do Útero/genética , Neoplasias do Colo do Útero/terapia
7.
Hum Gene Ther ; 14(15): 1389-99, 2003 Oct 10.
Artigo em Inglês | MEDLINE | ID: mdl-14577920

RESUMO

Interleukin (IL)-12 has been reported to induce cellular immune responses for protection against tumor formation. Here we investigate the utility of adenoviral delivery of IL-12 as an adjuvant for a human papillomavirus E7 subunit vaccine in a mouse tumor challenge model. Direct intratumoral injection of AdIL-12 resulted in a significant suppression of tumor growth compared to the control group. Injection of E7 protein into either a tumor site or the distance site along with AdIL-12 further enhanced antitumor effects significantly higher than either AdIL-12 or E7 injection alone. This combined injection resulted in complete regression of 9-mm-sized tumor in 40% of animals as well as lasting antitumor immunity against tumor recurrence. We also evaluated immune responses induced by these injections. AdIL-12 plus E7 enhanced E7-specific antibody responses significantly higher than AdIL-12 or E7 injection. In particular, the production level of interferon (IFN)-gamma from E7-specific CD4(+) T cells was similar between AdIL-12 group and AdIL-12 + E7 group. However, IFN-gamma production from E7-specific CD8(+) T cells was the most significant when injected with AdIL-12 + E7. This was consistent with intracellular IFN-gamma staining levels of CD8(+) T cells, suggesting that AdIL-12 + E7 injection enhances antitumor immunity in the human papillomavirus (HPV) 16 tumor model through increased expansion of the cytotoxic T-lymphocyte (CTL) subset. This enhanced protection appeared to be mediated by CD8(+) T cells, as determined by in vivo T-cell subset deletion. Thus, these studies demonstrate that E7 vaccines can induce CTL responses responsible for antitumor effects in the presence of IL-12 delivered via adenovirus vectors. This likely provides one additional approach for immune therapy against cervical cancers.


Assuntos
Adenoviridae/genética , Interleucina-12/genética , Proteínas Oncogênicas Virais/genética , Proteínas Repressoras , Neoplasias do Colo do Útero/terapia , Animais , Linfócitos T CD4-Positivos/metabolismo , Linfócitos T CD8-Positivos/metabolismo , Separação Celular , Citocinas/biossíntese , Dimerização , Modelos Animais de Doenças , Ensaio de Imunoadsorção Enzimática , Feminino , Citometria de Fluxo , Deleção de Genes , Terapia Genética , Humanos , Interferon gama/metabolismo , Camundongos , Camundongos Endogâmicos C57BL , Proteínas Oncogênicas Virais/uso terapêutico , Proteínas E7 de Papillomavirus , Proteínas Recombinantes/metabolismo , Baço/citologia , Fatores de Tempo
8.
DNA Cell Biol ; 22(3): 217-24, 2003 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-12804120

RESUMO

A constituent of green tea, (-)-epigallocatechin-3-gallate (EGCG) has been known to possess antiproliferative properties. In this study, we investigated the anticancer effects of EGCG in human papillomavirus (HPV)-16 associated cervical cancer cell line, CaSki cells. The growth inhibitory mechanism(s) and regulation of gene expression by EGCG were also evaluated. EGCG showed growth inhibitory effects in CaSki cells in a dose-dependent fashion, with an inhibitory dose (ID)(50) of approximately 35 microM. When CaSki cells were further tested for EGCG-induced apoptosis, apoptotic cells were significantly observed after 24 h at 100 microM EGCG. In contrast, an insignificant induction of apoptotic cells was observed at 35 microM EGCG. However, cell cycles at the G1 phase were arrested at 35 microM EGCG, suggesting that cell cycle arrests might precede apoptosis. When CaSki cells were tested for their gene expression using 384 cDNA microarray, an alteration in the gene expression was observed by EGCG treatment. EGCG downregulated the expression of 16 genes over time more than twofold. In contrast, EGCG upregulated the expression of four genes more than twofold, suggesting a possible gene regulatory role of EGCG. This data supports that EGCG can inhibit cervical cancer cell growth through induction of apoptosis and cell cycle arrest as well as regulation of gene expression in vitro. Furthermore, in vivo antitumor effects of EGCG were also observed. Thus, EGCG likely provides an additional option for a new and potential drug approach for cervical cancer patients.


Assuntos
Antineoplásicos Fitogênicos/farmacologia , Apoptose/efeitos dos fármacos , Carcinoma/tratamento farmacológico , Catequina/análogos & derivados , Catequina/farmacologia , Fase G1/efeitos dos fármacos , Neoplasias do Colo do Útero/tratamento farmacológico , Animais , Camellia sinensis/química , Testes de Carcinogenicidade , Carcinoma/genética , Carcinoma/patologia , Carcinoma/virologia , Divisão Celular/efeitos dos fármacos , Relação Dose-Resposta a Droga , Ensaios de Seleção de Medicamentos Antitumorais , Feminino , Citometria de Fluxo , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Humanos , Camundongos , Camundongos Nus , Análise de Sequência com Séries de Oligonucleotídeos , Papillomaviridae , Células Tumorais Cultivadas , Infecções Tumorais por Vírus/tratamento farmacológico , Infecções Tumorais por Vírus/patologia , Infecções Tumorais por Vírus/virologia , Neoplasias do Colo do Útero/genética , Neoplasias do Colo do Útero/patologia , Neoplasias do Colo do Útero/virologia
9.
Oncol Rep ; 12(3): 573-80, 2004 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-15289840

RESUMO

An arsenical compound, As2O3 has been reported to be effective for treating acute leukemia and induce apoptosis in many different tumor cell types. In this study we designed a novel arsenical compound, As4O6, and compared its ability to induce cell growth inhibition as well as gene expression profiles along with As2O3 in HPV16 infected SiHa cervical cancer cells. Both As2O3 and As4O6 induced apoptosis in SiHa cells, as determined by a DNA ladder formation. As4O6 was more effective in suppressing the growth of SiHa cells in vitro, as compared to As2O3. To further compare gene expression profiles between these two drugs, we used a 384 cDNA microarray system. The gene expression profiles were also classified into the Gene Ontology (GO) to investigate apoptosis-related cellular processes. In the case of As2O3, 41 genes were up- or down-regulated at least 2-fold, as compared to non-treatment, whereas, 65 genes were up- or down-regulated by As4O6 treatment. In particular, 27 genes were commonly regulated by both arsenic compounds. The GO analysis also indicated that down-regulation of cell-regulatory functions, such as cell cycle, protein kinase activity and DNA repair, induces an anti-tumor effect. Taken together, these data support that As4O6 could be more effective than As2O3 in inhibiting the growth of HPV16 infected cervical cancer cells. This appears to be mediated through a unique but overlapping regulatory mechanism(s), suggesting that the regulated genes and cellular processes could be used for a new potential drug approach for treating cervical cancer in clinical settings.


Assuntos
Arsenicais/uso terapêutico , Regulação Neoplásica da Expressão Gênica , Análise de Sequência com Séries de Oligonucleotídeos/métodos , Óxidos/uso terapêutico , Antineoplásicos/uso terapêutico , Apoptose , Trióxido de Arsênio , Northern Blotting , Ciclo Celular , Divisão Celular/efeitos dos fármacos , Linhagem Celular Tumoral , DNA/química , Fragmentação do DNA , Reparo do DNA , DNA Complementar/metabolismo , Regulação para Baixo , Feminino , Humanos , Hibridização de Ácido Nucleico , RNA/química , Fatores de Tempo , Regulação para Cima , Neoplasias do Colo do Útero/tratamento farmacológico
10.
Int J Oncol ; 43(2): 539-47, 2013 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-23707988

RESUMO

There is an urgent need for molecular marker studies of adenocarcinoma (AC) and squamous cell carcinoma (SCC) of the uterine cervix. This study utilized oligomicroarray and pathway analyses to characterize a transcriptomic signature with molecular networks associated with AC and SCC. A 10K oligomicroarray was used to identify potential transcripts that were differentially expressed in cervical cancers from 28 patients and common reference RNAs from 17 different normal cervixes. Molecular networks were correlated using genomics tools to globally explore cellular pathways. Gene expression levels of 46 transcripts separated cancer samples into AC and SCC groups. Genes including: KRT17, IGFBP2, CALCA and VIPR1 were differentially expressed in AC and SCC. In addition, we identified a transcriptomic signature that predicted tumor classification and progression based upon its cellular processes. The downregulated signatures for SCC were cell death of pheochromocytoma cells (P=0.0037), apoptosis of neurons (P=0.009) and damage to DNA (P=0.0038). By contrast, the upregulated molecular signatures in AC were immunological disorder (P=0.006), splenomegaly (P=0.0053) and hepatic system disorder (P=0.006). The G2/M DNA damage checkpoint regulation pathway (P=0.05) was found to be significantly linked to IGF1R as a new regulatory component of a putative cytoplasmic signaling cascade in SCC. By contrast, the antigen presenting canonical pathway (P=0.038) appeared to be linked to PPARγ in AC. Taken together, these experiments provide important new information regarding the role of molecular networks in mediating SCC and AC, possibly through two independent pathways, and contribute to provide new targets for the prevention and treatment of cervical cancer.


Assuntos
Adenocarcinoma/genética , Biomarcadores Tumorais/genética , Carcinoma de Células Escamosas/genética , Neoplasias do Colo do Útero/genética , Calcitonina/genética , Peptídeo Relacionado com Gene de Calcitonina , Regulação para Baixo , Feminino , Pontos de Checagem da Fase G2 do Ciclo Celular , Humanos , Proteína 2 de Ligação a Fator de Crescimento Semelhante à Insulina/genética , Queratina-17/genética , PPAR gama/genética , Precursores de Proteínas/genética , Receptor IGF Tipo 1/genética , Receptores Tipo I de Polipeptídeo Intestinal Vasoativo/genética , Transdução de Sinais/genética , Regulação para Cima
11.
PLoS One ; 7(6): e38583, 2012.
Artigo em Inglês | MEDLINE | ID: mdl-22715393

RESUMO

The effects of As(4)O(6) as adjuvant on photodynamic therapy (PDT) were studied. As(4)O(6) is considered to have anticancer activity via several biological actions, such as free radical production and inhibition of VEGF expression. PDT or As(4)O(6) significantly inhibited TC-1 cell proliferation in a dose-dependent manner (P<0.05) by MTT assay. The anti-proliferative effect of the combination treatment was significantly higher than in TC-1 cells treated with either photodynamic therapy or As(4)O(6) alone (62.4 and 52.5% decrease compared to vehicle-only treated TC-1 cells, respectively, P<0.05). In addition, cell proliferation in combination of photodynamic therapy and As(4)O(6) treatment significantly decreased by 77.4% (P<0.05). Cell survival pathway (Naip1, Tert and Aip1) and p53-dependent pathway (Bax, p21(Cip1), Fas, Gadd45, IGFBP-3 and Mdm-2) were markedly increased by combination treatment of photodynamic therapy and As(4)O(6). In addition, the immune response in the NEAT pathway (Ly-12, CD178 and IL-2) was also modulated after combination treatment, suggesting improved antitumor effects by controlling unwanted growth-stimulatory pathways. The combination effect apparently reflected concordance with in vitro data, in restricting tumor growth in vivo and in relation to some common signaling pathways to those observed in vitro. These findings suggest the benefit of combinatory treatment with photodynamic therapy and As(4)O(6) for inhibition of cervical cancer cell growth.


Assuntos
Arsenicais/farmacologia , Neoplasias Experimentais/tratamento farmacológico , Óxidos/farmacologia , Fotoquimioterapia/métodos , Neoplasias do Colo do Útero/tratamento farmacológico , Animais , Trióxido de Arsênio , Linhagem Celular Tumoral , Ensaios de Seleção de Medicamentos Antitumorais/métodos , Feminino , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Camundongos , Proteínas de Neoplasias/biossíntese , Neoplasias Experimentais/metabolismo , Neoplasias Experimentais/patologia , Neoplasias do Colo do Útero/metabolismo , Neoplasias do Colo do Útero/patologia
12.
PLoS One ; 7(9): e44960, 2012.
Artigo em Inglês | MEDLINE | ID: mdl-22970327

RESUMO

CA125 as a biomarker of ovarian cancer is ineffective for the general population. The aim of this study was to evaluate multiplexed bead-based immunoassay of multiple ovarian cancer-associated biomarkers such as transthyretin and apolipoprotein A1, together with CA125, to improve the identification and evaluation of prognosis of ovarian cancer. We measured the serum levels of CA125, transthyretin, and apolipoprotein A1 from the serum of 61 healthy individuals, 84 patients with benign ovarian disease, and 118 patients with ovarian cancer using a multiplex liquid assay system, Luminex 100. The results were then analyzed according to healthy and/or benign versus ovarian cancer subjects. When CA125 was combined with the other biomarkers, the overall sensitivity and specificity were significantly improved in the ROC curve, which showed 95% and 97% sensitivity and specificity, respectively. At 95% specificity for all stages the sensitivity increased to 95.5% compared to 67% for CA125 alone. For stage I+II, the sensitivity increased from 30% for CA125 alone to 93.9%. For stage III+IV, the corresponding values were 96.5% and 91.6%, respectively. Also, the three biomarkers were sufficient for maximum separation between noncancer (healthy plus benign group) and stage I+II or all stages (I-IV) of disease. The new combination of transthyretin, and apolipoprotein A1 with CA125 improved both the sensitivity and the specificity of ovarian cancer diagnosis compared with those of individual biomarkers. These findings suggest the benefit of the combination of these markers for the diagnosis of ovarian cancer.


Assuntos
Biomarcadores Tumorais/sangue , Imunoensaio/métodos , Neoplasias Ovarianas/diagnóstico , Estudos de Casos e Controles , Diagnóstico Precoce , Feminino , Humanos , Neoplasias Ovarianas/sangue , Curva ROC , Sensibilidade e Especificidade
13.
PLoS One ; 7(2): e32259, 2012.
Artigo em Inglês | MEDLINE | ID: mdl-22393393

RESUMO

The accurate genotyping of human papillomavirus (HPV) is clinically important because the oncogenic potential of HPV is dependent on specific genotypes. Here, we described the development of a bead-based multiplex HPV genotyping (MPG) method which is able to detect 20 types of HPV (15 high-risk HPV types 16, 18, 31, 33, 35, 39, 45, 51, 52, 53, 56, 58, 59, 66, 68 and 5 low-risk HPV types 6, 11, 40, 55, 70) and evaluated its accuracy with sequencing. A total of 890 clinical samples were studied. Among these samples, 484 were HPV positive and 406 were HPV negative by consensus primer (PGMY09/11) directed PCR. The genotyping of 484 HPV positive samples was carried out by the bead-based MPG method. The accuracy was 93.5% (95% CI, 91.0-96.0), 80.1% (95% CI, 72.3-87.9) for single and multiple infections, respectively, while a complete type mismatch was observed only in one sample. The MPG method indiscriminately detected dysplasia of several cytological grades including 71.8% (95% CI, 61.5-82.3) of ASCUS (atypical squamous cells of undetermined significance) and more specific for high grade lesions. For women with HSIL (high grade squamous intraepithelial lesion) and SCC diagnosis, 32 women showed a PPV (positive predictive value) of 77.3% (95% CI, 64.8-89.8). Among women >40 years of age, 22 women with histological cervical cancer lesions showed a PPV of 88% (95% CI, 75.3-100). Of the highest risk HPV types including HPV-16, 18 and 31 positive women of the same age groups, 34 women with histological cervical cancer lesions showed a PPV of 77.3% (95% CI, 65.0-89.6). Taken together, the bead-based MPG method could successfully detect high-grade lesions and high-risk HPV types with a high degree of accuracy in clinical samples.


Assuntos
Técnicas Genéticas , Papillomaviridae/genética , Infecções por Papillomavirus/diagnóstico , Infecções por Papillomavirus/genética , Adulto , Idoso , Feminino , Genes Virais , Genótipo , Humanos , Pessoa de Meia-Idade , Modelos Genéticos , Hibridização de Ácido Nucleico , Reprodutibilidade dos Testes , Sensibilidade e Especificidade , Análise de Sequência de DNA , Especificidade da Espécie , Neoplasias do Colo do Útero/diagnóstico , Neoplasias do Colo do Útero/genética
14.
J Photochem Photobiol B ; 110: 50-7, 2012 May 02.
Artigo em Inglês | MEDLINE | ID: mdl-22487363

RESUMO

Chlorin-based photosensitizers in photodynamic therapy are the promising anticancer agents, but some of their properties such as specific-targeting to tumor need to be improved. The aim of this study was to synthesize chlorin-based unsaturated fatty acid conjugates to obtain an optimal photosensitizers. Thus four chlorin-based fatty acid conjugates were successfully synthesized through an esterification reaction using carbodiimide coupling reagents in enough yields. Then, structures of these conjugates were confirmed by (1)H NMR, MALDI-MS, and UV-vis spectroscopy. Furthermore, their in vitro phototoxicity and cellular uptake were evaluated on TC-1 lung cancer cell line and HeLa cell line.


Assuntos
Ácidos Graxos Insaturados/química , Fármacos Fotossensibilizantes/síntese química , Porfirinas/síntese química , Linhagem Celular Tumoral , Ácidos Graxos Insaturados/uso terapêutico , Células HeLa , Humanos , Neoplasias Pulmonares/tratamento farmacológico , Ressonância Magnética Nuclear Biomolecular , Fotoquimioterapia , Fármacos Fotossensibilizantes/uso terapêutico , Porfirinas/uso terapêutico
15.
PLoS One ; 7(5): e37772, 2012.
Artigo em Inglês | MEDLINE | ID: mdl-22662218

RESUMO

BACKGROUND: We previously reported that the USP17 deubiquitinating enzyme having hyaluronan binding motifs (HABMs) interacts with human SDS3 (suppressor of defective silencing 3) and specifically deubiquitinates Lys-63 branched polyubiquitination of SDS3 resulting in negative regulation of histone deacetylase (HDAC) activity in cancer cells. Furthermore, USP17 and SDS3 mutually interact with each other to block cell proliferation in HeLa cells but the mechanism for this inhibition in cell proliferation is not known. We wished to investigate whether the HABMs of USP17 were responsible for tumor suppression activity. METHODOLOGY/PRINCIPAL FINDINGS: Similarly to USP17, we have identified that SDS3 also has three consecutive HABMs and shows direct binding with hyaluronan (HA) using cetylpyridinium chloride (CPC) assay. Additionally, HA oligosaccharides (6-18 sugar units) competitively block binding of endogenous HA polymer to HA binding proteins. Thus, administration of HA oligosaccharides antagonizes the interaction between HA and USP17 or SDS3. Interestingly, HABMs deleted USP17 showed lesser interaction with SDS3 but retain its deubiquitinating activity towards SDS3. The deletion of HABMs of USP17 could not alter its functional regulation on SDS3-associated HDAC activity. Furthermore, to explore whether HABMs in USP17 and SDS3 are responsible for the inhibition of cell proliferation, we investigated the effect of USP17 and SDS3-lacking HABMs on cell proliferation by soft agar, apoptosis, cell migration and cell proliferation assays. CONCLUSIONS: Our results have demonstrated that these HABMs in USP17 and its substrate SDS3 are mainly involved in the inhibition of anchorage-independent tumor growth.


Assuntos
Endopeptidases/química , Endopeptidases/metabolismo , Ácido Hialurônico/metabolismo , Domínios e Motivos de Interação entre Proteínas , Proteínas Repressoras/química , Proteínas Repressoras/metabolismo , Sequência de Aminoácidos , Sequência de Bases , Proliferação de Células , Endopeptidases/genética , Ativação Enzimática/genética , Células HeLa , Humanos , Dados de Sequência Molecular , Neoplasias/genética , Neoplasias/metabolismo , Ligação Proteica , Proteínas Repressoras/genética , Deleção de Sequência
16.
Oncol Rep ; 28(2): 576-84, 2012 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-22614712

RESUMO

Selenium, an essential trace element possessing anti-carcinogenic properties, can induce apoptosis in cancer cells. Our goal was to investigate the enhanced antitumor effects of photodynamic therapy (PDT) plus selenium in TC-1 tumor cells and implanted mice. Cell viability was evaluated at various time intervals after PDT treatment and/or selenium by methyl thiazolyl tetrazolium (MTT) assay. When only PDT treatment was administered to TC-1 tumor cells, TC-1 cell growth recovered over time. On the other hand, co-treatment of PDT and selenium extended the inhibition time of tumor cell growth. Co-treatment of PDT and selenium showed serious morphological changes in TC-1 cells and induced a more apoptotic population by FACS analysis. By signal transduction pathway SuperArray analysis, genes closely involved in the NFκB, p53 and phopholipase C pathways, such as VCAM1, MDM2 and FOS, were significantly downregulated at least 10-fold in TC-1 cells following PDT and selenium cotreatment. In an in vivo study, tumor-bearing mice were intravenously injected with Radachlorin 3 h before irradiation with 300 J/cm2 of light. Selenium was administered daily for 20 days. Combination therapy against the mouse tumors generated by TC-1 cells was more effective than PDT or selenium alone. These data suggest that selenium plus PDT can induce a significant tumor suppression response compared with PDT alone. Additionally, it can be an effective anticancer therapy strategy.


Assuntos
Neoplasias Experimentais/tratamento farmacológico , Fotoquimioterapia/métodos , Fármacos Fotossensibilizantes/farmacologia , Porfirinas/farmacologia , Selênio/farmacologia , Animais , Processos de Crescimento Celular/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Sinergismo Farmacológico , Feminino , Expressão Gênica , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Nus , Neoplasias Experimentais/metabolismo , Neoplasias Experimentais/patologia
17.
Biol Trace Elem Res ; 148(1): 25-31, 2012 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-22328307

RESUMO

The aim of this study was to determine serum selenium (Se) levels during the development of liver disease as well as the possible Se supplementation benefits in liver disease patients. Serum was collected from 187 patients with liver diseases and 120 normal healthy people living in Seoul. The samples were collected at the Kangnam St. Mary's Hospital College of Medicines, The Catholic University of Korea, in accordance with procedures approved by the Institutional Review Board of the Catholic University of Korea. Serum Se levels were quantified by inductively coupled plasma mass spectrometry and were compared between healthy and liver diseases patients. Se levels were 92.65 ± 32.50 µg/l in hepatitis infection, 92.33 ± 30.66 µg/l in hepatitis B virus infection and 96.41 ± 51.50 µg/l in hepatitis C virus infection, 96.42 ± 32.80 µg/l in cirrhosis, and 67.47 ± 14.30 µg/l in hepatoma patients. Findings were significantly lower in hepatitis and hepatoma as compared with the healthy participants (P < 0.001). The Se level of the healthy population was 108.38 ± 29.50, 119.37 ± 28.31 for males and 97.87 ± 26.99 µg/l for females. Our data shows the same parallelism between liver disease progression and decrease of Se levels except in the case of liver cirrhosis. And also, our study confirms the previous findings of significantly lower Se levels in Korean hepatoma patients. Se levels that decrease parallel to liver disease progression should be further integrated and analyzed with liver function blood biomarkers.


Assuntos
Biomarcadores Tumorais/sangue , Carcinoma Hepatocelular/sangue , Selênio/sangue , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Povo Asiático , Carcinoma Hepatocelular/epidemiologia , Feminino , Hepatite/sangue , Hepatite/epidemiologia , Humanos , Cirrose Hepática/sangue , Cirrose Hepática/epidemiologia , Masculino , Pessoa de Meia-Idade , República da Coreia/epidemiologia
18.
J Cancer Res Clin Oncol ; 138(7): 1173-86, 2012 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-22419440

RESUMO

OBJECTIVE: The aim of this study was to identify novel genes following genomic DNA copy number changes using a genome-wide array-based comparative genomic hybridization (array-CGH) analysis in uterine leiomyosarcoma (ULMS). METHODS: Genomic DNA copy number changes were analyzed in 15 cases of ULMS from St Mary's Hospital of the Catholic University of Korea. The paraffin-fixed tissue samples were micro-dissected under microscope, and DNA was extracted. Array-based CGH and genomic polymerase chain reaction were carried out with statistical analyses such as hierarchical clustering and Gene Ontology. RESULTS: All of 15 cases of ULMS showed specific gains and losses. The percentage of average gains and losses were 8.4 and 16.6 %, respectively. The analysis limit of average gains and losses was 40 %. The regions of high level of gain were 1q23.3, 7p14.2, 7q34, 7q35, 7q36.3, 13q34, and 16p13.3. And the regions of homozygous loss were 2q21.1, 2q22.1, 2p23.2, 12q23.3, 4q21.22, 4q34.3, 11q24.2, 12q23.3, 13q13.1, 13q21.33, and 14q24.3. In ULMS samples, recurrent regions of gain were 1p36.33, 1p36.32, 5q35.3, 7q36.3, and 8q24.3 and recurrent regions of loss were 1p31.1-p31.3, 1p32.1-p32.3, 2p12, 2p13.3, 2p14, 2p16.2-p16.3, 2q12.1-q12.3, 2q21.1-q21.2, 2q22.2-q22.3, 2q34, 2q36.1-q36.3, 5q21.3, 5q23.3, 5q31.1, 6p11.2, 6p12.1, 10q11.23, 10q21.2-q21.3, 10q23.2, 10q23.31, 10q25.1-q25.2, 10q25.3, 10q26.13, 10q26.2-q26.3, 11p11.2, 11p11.12, 11p12, 11p13, 11p15.4, 11q23.1-q23.2, 11q23.3, 13q14.12, 13q14.13-13q14.2, 13q14.2, 13q14.2, 13q14.3, 13q21.33, 13q22.1-q22.3, 14q24.2, 14q24.3, 14q31.1, 14q32.33, 15q11.2-q13, 15q14, 16q22.3, 16q23.1, 16q23.2, 16q24.1, 20p12.1, and 21q22.3. Representative frequently gained BAC clones encoded genes were HDAC9, CRR9, SOX18, PTPRN2, SKI, SOLH, and KIAA1199. The genes encoded by frequently lost BAC clones were LOC150516 and AMY2A. A subset of cellular processes from each gene were clustered by Gene Ontology database. CONCLUSIONS: The present study using array-CGH analyses sought a deeper elucidation of the specific genomic alterations related to ULMS. The high resolution of array-CGH combined with human genome database would give a chance at identifying relevant target genes.


Assuntos
Aberrações Cromossômicas , Hibridização Genômica Comparativa , Leiomiossarcoma/genética , Neoplasias Uterinas/genética , Adulto , Bases de Dados Genéticas , Feminino , Dosagem de Genes , Perfilação da Expressão Gênica , Genoma Humano , Humanos , Leiomiossarcoma/patologia , Pessoa de Meia-Idade , Neoplasias Uterinas/patologia
19.
Oncol Rep ; 28(2): 585-91, 2012 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-22641176

RESUMO

The purpose of the present study was to evaluate multiplex liquid assay-based measurement of multiple ovarian cancer-associated biomarkers such as hemoglobin, haptoglobin and apolipoprotein E, together with CA125, which has been widely used in the diagnosis of ovarian cancer, in order to provide a higher diagnostic power. We measured the serum levels of CA125, hemoglobin, haptoglobin and apolipoprotein E from the serum of 76 healthy individuals and 69 ovarian cancer patients using a multiplex liquid assay system, Luminex 100. The results were analyzed according to normal versus ovarian cancer, tumor stages and tumor histology. In addition, to validate the use of these biomarkers for the diagnosis of ovarian cancer, the sensitivity and specificity of each biomarker was analyzed by its receiver operating characteristics (ROC) curve. The serum levels of all four biomarkers in ovarian cancer patients were significantly higher than those of healthy individuals. When CA125 was combined with the biomarkers, the overall sensitivity and specificity were significantly improved in the ROC curve, which showed 95 and 75% sensitivity and specificity, respectively. At 95% specificity for all stages the sensitivity increased to 75% compared to 41% for CA125 alone. For stage I+II increased the sensitivity to 68% from 36% for CA125 alone. For stage III+IV the corresponding values were 100 and 95%, respectively. Taken together, the new combination of hemoglobin, haptoglobin and apolipoprotein E with CA125 significantly improved both the sensitivity and the specificity of ovarian cancer diagnosis compared with those of individual biomarkers. These findings suggest the benefit of the combination of these markers for the diagnosis of ovarian cancer.


Assuntos
Biomarcadores Tumorais/sangue , Imunoensaio/métodos , Neoplasias Ovarianas/sangue , Neoplasias Ovarianas/diagnóstico , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Antígeno Ca-125/sangue , Estudos de Casos e Controles , Feminino , Humanos , Pessoa de Meia-Idade , Neoplasias Ovarianas/patologia , Análise de Sobrevida , Adulto Jovem
20.
Int J Oncol ; 41(6): 2038-46, 2012 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-23023522

RESUMO

Host genomic alterations in addition to human papillomavirus (HPV) are needed for cervical precursor lesions to progress to invasive cancer because only a small percentage of women infected by the virus develop disease. However, the genomic alterations during the progression of cervical lesions have not been systematically examined. The aim of this study was to identify differential genomic alterations among cervical intraepithelial neoplasia CIN1, CIN2, CIN3 and cervical squamous cell carcinoma (SCC). Genomic alterations were examined for 15 cases each of CIN1, CIN2, CIN3 and SCC by array-based comparative genomic hybridization (array CGH). The chromosomal regions showing significant differential in DNA copy number aberrations (DCNAs) among CIN1, CIN2, CIN3 and SCC were successfully identified by resampling-based t-test. The chromosomal regions of 5q35.3 and 2q14.3 showed significant DCNAs between CIN1 and CIN2, and between CIN2 and CIN3, respectively, while a significant difference in DCNAs between CIN3 and SCC was observed at 1q24.3, 3p14.1, 3p14.2, 5q13.2, 7p15.3, 7q22.1 and 13q32.3. In addition, the status of DCNAs in 1q43, 2p11.2, 6p11.2, 7p21.1, 7p14.3, 10q24.1, 13q22.3, 13q34 and 16p13.3 was conserved throughout the progression of CIN to SCC. The presence of differential and common DCNAs among CIN1, CIN2, CIN3 and SCC supports that the CIN progression may include continual clonal selection and evolution. This approach also identified 34 probe sets consistently overexpressed when amplified, suggesting an unbiased identification of candidate genes in SCC during cervical cancer progression.


Assuntos
Variações do Número de Cópias de DNA , Displasia do Colo do Útero/genética , Displasia do Colo do Útero/patologia , Neoplasias do Colo do Útero/genética , Neoplasias do Colo do Útero/patologia , Adulto , Aberrações Cromossômicas , Análise por Conglomerados , Hibridização Genômica Comparativa , Progressão da Doença , Feminino , Humanos , Pessoa de Meia-Idade , Invasividade Neoplásica , Estadiamento de Neoplasias , Infecções por Papillomavirus/virologia , Neoplasias do Colo do Útero/virologia , Displasia do Colo do Útero/virologia
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