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1.
Nanobiomedicine (Rij) ; 6: 1849543519880762, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31908670

RESUMO

The purpose of this study was to investigate the efficacy of targeting peptides chemotherapy to overcome adverse event in the conventional chemotherapy for human hepatocellular carcinoma. Previously we reported several cancer-targeting peptides that bind specifically to cancer cells and their vascular endothelia: L-peptide (anti-cancer cell membrane), RLLDTNRPLLPY; SP-94-peptide (anti-hepatoma cell membrane), SFSHHTPILP; PC5-52-peptide (anti-tumor endothelia), SVSVGMKPSPRP; and control peptide, RLLDTNRGGGGG. In this study, these peptides were linked to liposomal iron oxide nanoparticles to localize the targeted tumor cells and endothelia, and to dextran-coated liposomal doxorubicin (L-D) to treat nonobese diabetic severe combined immunodeficient mice bearing hepatoma xenografts. Our results showed that L-peptide-linked liposomal doxorubicin could inhibit tumor growth with very mild adverse events. Use of the control peptide led to a decrease in the xenograft size but also led to marked apoptotic change in the visceral organ. In conclusion, L-peptide-linked liposomal doxorubicin, SP-94-peptide, and PC5-52-peptide can be used for the treatment of hepatoma xenografts in nonobese diabetic severe combined immunodeficient mice with minimal adverse events.

2.
Int J Oncol ; 53(3): 1105-1117, 2018 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-29956746

RESUMO

Paclitaxel (PTX) exhibits potent antineoplastic activity against various human malignancies; however, clinical application must overcome the inherent hydrophobicity of this molecule. The commercialized Taxol formulation utilizes Cremophor EL (CrEL)/ethanol as a solvent to stabilize and dispense PTX in an aqueous solution. However, adverse CrEL­induced hypersensitivity reactions have been reported in ~30% of recipients, and 40% of patients receiving premedication may also experience this adverse effect. Therefore, the development of a CrEL-free delivery system is crucial, in order to fully exploit the therapeutic efficacy of PTX. In the present study, a novel liposomal PTX (lipo­PTX) formulation was optimized with regards to encapsulation rate and long­term stability, arriving at a molar constituent ratio of soybean phosphatidylcholine:cholesterol:N-(carbonyl-methoxy-poly-ethylene glycol 2000)­1,2­distearoyl­sn-glycero­3-phosphoethanolamine, sodium salt:PTX at 95:2:1:2. Comparable doses of lipo­PTX and Taxol were bioequivalent in terms of therapeutic efficacy in xenograft tumor models. However, the systemic side effects, including hematopoietic toxicity, acute hypersensitivity reactions and cardiac irregularities, were significantly reduced in lipo­PTX­treated mice compared with those infused with reference formulations of PTX. In conclusion, the present study reported that lipo­PTX exhibited a higher therapeutic index than clinical PTX formulations.


Assuntos
Antineoplásicos Fitogênicos/farmacologia , Portadores de Fármacos/efeitos adversos , Composição de Medicamentos/métodos , Paclitaxel/farmacologia , Animais , Antineoplásicos Fitogênicos/uso terapêutico , Doenças Cardiovasculares/induzido quimicamente , Doenças Cardiovasculares/epidemiologia , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos dos fármacos , Portadores de Fármacos/química , Liberação Controlada de Fármacos , Ensaios de Seleção de Medicamentos Antitumorais , Feminino , Glicerol/efeitos adversos , Glicerol/análogos & derivados , Glicerol/química , Doenças Hematológicas/induzido quimicamente , Doenças Hematológicas/epidemiologia , Humanos , Lipossomos , Masculino , Camundongos , Camundongos Endogâmicos ICR , Camundongos SCID , Neutropenia , Paclitaxel/uso terapêutico , Equivalência Terapêutica , Distribuição Tecidual
3.
Nanomedicine ; 11(6): 1425-34, 2015 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-25881740

RESUMO

The L-peptide plays a role as a universal ligand binding specifically to nasopharyngeal carcinoma (NPC) and other cancers but not normal cells. It was used to link iron oxide nanoparticles, and injected intravenously to SCID mice bearing NPC and breast cancer xenografts for MR analysis, and showed significant change of MR signal intensity in the xenograft regions. Using this conjugate as a ligand to localize the L-peptide targeted protein in the cancer surgical specimens, a clear reaction product was identified in the tumor cells of both cancer types. If the L-peptide-linked-liposomal doxorubicin was used to treat the SCID mice bearing other NPC or breast cancer xenograft, a high efficacy of chemotherapy with minimal adverse effect was observed. In conclusion, the L-peptide has a considerable potential for clinical usage for targeted imaging, peptide histochemical localization of targeted protein, and targeted chemotherapy for different cancer types. FROM THE CLINICAL EDITOR: Targeted chemotherapy to cancer cells will enable maximum drug delivery but minimal systemic side effects. In this article, the authors identified a protein, L-peptide, on tumor cells. They also subsequently confirmed the specificity of this protein in animal experiments using iron oxide nanoparticles. The discovery of this marker could lead to future development of better chemotherapy.


Assuntos
Neoplasias da Mama/diagnóstico , Neoplasias da Mama/tratamento farmacológico , Nanopartículas , Neoplasias Nasofaríngeas/diagnóstico , Neoplasias Nasofaríngeas/tratamento farmacológico , Peptídeos/uso terapêutico , Animais , Linhagem Celular Tumoral , Imageamento por Ressonância Magnética , Camundongos , Camundongos SCID , Peptídeos/administração & dosagem , Ensaios Antitumorais Modelo de Xenoenxerto
4.
J Biomater Appl ; 28(4): 580-94, 2013 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-23174955

RESUMO

The purpose of this study was to analyze the encapsulation of superparamagnetic iron oxide nanoparticles (SPION) by the lipid nanoparticle conjugated with the 12-mer peptides (RLLDTNRPLLPY, L-peptide), and the delivery of this complex into living cells. The lipid nanoparticles employed in this work were highly hydrophilic, stable, and contained poly(ethylene-glycol) for conjugation to the bioactive L-peptide. The particle sizes of two different magnetic lipid nanoparticles, L-peptide modified (LML) and non-L-peptide modified (ML), were both around 170 nm with a narrow range of size disparity. The transversal relaxivity, r2, for both LML and ML nanoparticles were found to be significantly higher than the longitudinal relaxivity r1 (r2/r1 > 20). The in vitro tumor cell targeting efficacy of the LML nanoparticles were evaluated and compared to the ML nanoparticles, upon observing cellular uptake of magnetic lipid nanoparticles by the nasopharyngeal carcinoma cells, which express cell surface specific protein for the L-peptide binding revealed. In the Prussian blue staining experiment, cells incubated with LML nanoparticles indicated much higher intracellular iron density than cells incubated with only the ML and SPION nanoparticles. In addition, the MTT assay showed the negligible cell cytotoxicity for LML, ML and SPION nanoparticles. The MR imaging studies demonstrate the better T2-weighted images for the LML-nanoparticle-loaded nasopharyngeal carcinoma cells than the ML- and SPION-loaded cells.


Assuntos
Meios de Contraste , Imageamento por Ressonância Magnética/métodos , Magnetismo , Nanopartículas , Neoplasias Nasofaríngeas/diagnóstico , Humanos , Técnicas In Vitro , Espectroscopia de Ressonância Magnética , Microscopia Eletrônica de Transmissão , Tamanho da Partícula , Difração de Pó
5.
Oncol Rep ; 27(1): 94-100, 2012 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-21956494

RESUMO

Nucleolin (C23, NCL) mRNA was up-regulated in nasopharyngeal carcinoma (NPC) cells compared to that of normal nasomucosal (NNM) cells using a cDNA microarray approach. The level of nucleolin protein was also up-regulated in 13 NPC cell lines, 30 biopsy specimens and nine other cancer cell lines compared to five NNM cells or normal stromal cells, which were analyzed using immunoblotting or immunohistochemistry. We transfected nucleolin antisense oligodeoxynucleotides (phosphorothioate-modified oligodeoxynucleotides; S-ODNs) into NPC-TW01 cells to knockdown nucleolin expression to evaluate the function of nucleolin in cancer cells. Nucleolin knockdown induced NPC cells but not NNM cells to undergo apoptosis. Furthermore, treatment of NPC-TW01 xenograft tumors with nucleolin antisense oligodeoxynucleotides suppressed the growth of xenograft tumors without obvious side effects. Therefore, we suggest that nucleolin may be a potential cancer therapeutic target and that nucleolin antisense oligodeoxynucleotides may be used as a potential drug for therapy in NPC.


Assuntos
Antineoplásicos/farmacologia , Apoptose/genética , Carcinoma/genética , Neoplasias Nasofaríngeas/genética , Oligodesoxirribonucleotídeos Antissenso/farmacologia , Fosfoproteínas/antagonistas & inibidores , Proteínas de Ligação a RNA/antagonistas & inibidores , Animais , Apoptose/efeitos dos fármacos , Linhagem Celular Tumoral , Feminino , Técnicas de Silenciamento de Genes , Humanos , Immunoblotting , Imuno-Histoquímica , Marcação In Situ das Extremidades Cortadas , Camundongos , Camundongos SCID , Oligodesoxirribonucleotídeos Antissenso/genética , Fosfoproteínas/genética , RNA Mensageiro/análise , Proteínas de Ligação a RNA/genética , Reação em Cadeia da Polimerase em Tempo Real , Transfecção , Ensaios Antitumorais Modelo de Xenoenxerto , Nucleolina
6.
J Biomed Sci ; 18: 80, 2011 Nov 07.
Artigo em Inglês | MEDLINE | ID: mdl-22059741

RESUMO

BACKGROUND: MiR-1 (microRNA-1) has been used as a positive control in some microRNA experiments. We found that miR-1 transfection of nasopharyngeal carcinoma cells reveals a typical apoptotic process as shown by time-lapse microscopy so we investigated the mechanisms of miR-1 inducing apoptosis. METHODS: To confirm that miR-1 induces apoptosis, we used Annexin V and TUNEL staining and caspase assay. To determine that miR-1 directly targets genes that involve in apoptosis, we analyzed microRNA and pathway databases, and cDNA expression microarrays from miR-1 transfected cells. To demonstrate candidate miR-1 targeted genes, we used qRT-PCR analysis and luciferase reporter vector assays. To assess the miR-1 target gene PTMA (prothymosin alpha, ProTalpha) involves in apoptosis, we used PTMA siRNA to knock down PTMA. RESULTS: Annexin V and TUNEL staining and caspase assay confirm that miR-1 induces nasopharyngeal carcinoma cell apoptosis. MiR-1 transfection of HeLa, Cal-27, KYSE30 and NPC-TW06 cell lines which express low levels of endogenous miR-1 also induces apoptosis. However, miR-1 transfection of cell lines such as SW620, HepG2, HEK-293T, SAS and PC-13 which express high levels of endogenous miR-1 does not result in apoptosis. MiR-1 directly targets PTMA gene. PTMA siRNA and miR-1 accelerate the apoptotic process in cells treated with apoptosis inducers. CONCLUSIONS: The exogenous expression of miR-1 induces apoptosis in a number of cell lines. This is a model of microRNA-induced cell apoptosis. The PTMA is one of miR-1 target genes which involve in miR-1 inducing apoptosis. The apoptotic inducers including actinomycin D, camptothecin and etoposide are also the chemotherapeutic drugs in clinical cancer therapy and PTMA siRNA can accelerate apoptotic progression in cells treated with those apoptosis inducers. Therefore PTMA siRNA may have potential applications as an adjuvant in cancer chemotherapy.


Assuntos
Apoptose , MicroRNAs/metabolismo , Neoplasias Nasofaríngeas/genética , Precursores de Proteínas/genética , Timosina/análogos & derivados , Carcinoma , Linhagem Celular Tumoral , Humanos , Carcinoma Nasofaríngeo , Neoplasias Nasofaríngeas/metabolismo , Neoplasias Nasofaríngeas/patologia , Precursores de Proteínas/metabolismo , Timosina/genética , Timosina/metabolismo , Transfecção
7.
J Pathol ; 222(3): 299-309, 2010 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-20635349

RESUMO

Nasopharyngeal carcinoma (NPC) is prevalent in south-eastern Asia, particularly southern China, Singapore and Taiwan. The aim of this study was to identify the pivotal genes that may be altered during NPC progression. Using cDNA microarray analysis, we compared the expression of 18 genes between NPC and normal nasomucosal cells. qRT-PCR analysis found the expression of IBFBP-6 in NPC cell lines and immunolocalization of IGFBP-6 in NPC to be very weak. To explore the effects of IGFBP-6 on NPC tumour growth, we constructed inducible plasmids containing full-length IGFBP-6 cDNA (pBIG2i-IGFBP-6) and established pBIG2i-IGFBP-6-transfected NPC stable cell lines (NPC-TW01-pBIG2i-IGFBP-6). We then performed functional analysis of the IGFBP-6 in cell lines in vitro and in vivo. Over-expression of IGFBP-6 significantly suppressed the proliferation, invasion and metastatic activity of NPC cells and increased their apoptosis. We found the EGR-1, caspase-1 and TSP-1 genes to be markedly up-regulated when NPC-pBIG2i-IGFBP-6 was treated with doxycycline. Knocking down EGR-1 with EGR-1 siRNA resulted in a decrease in expression of caspase-1, TSP-1 and EGR-1 but not the expression of IGFBP-6. However, in knockdown cells the unchanged expression of IGFBP-6 did not inhibit the migration of NPC cells. Chromatin immunoprecipitation and luciferase reporter assay experiments showed that IGFBP-6 bound the EGR-1 promoter regions and activated EGR-1 promoter. We conclude that IGFBP-6 can regulate the progression of NPC by regulating the expression of EGR-1. These results suggest that IGFBP-6 could be used as a new target in NPC therapy.


Assuntos
Proteína 1 de Resposta de Crescimento Precoce/metabolismo , Genes Supressores , Proteína 6 de Ligação a Fator de Crescimento Semelhante à Insulina/genética , Neoplasias Nasofaríngeas/genética , Animais , Biópsia , Regulação Neoplásica da Expressão Gênica , Humanos , Proteína 6 de Ligação a Fator de Crescimento Semelhante à Insulina/metabolismo , Camundongos , Camundongos SCID , Neoplasias Nasofaríngeas/metabolismo , Neoplasias Nasofaríngeas/patologia , Proteínas de Neoplasias/metabolismo , Regiões Promotoras Genéticas , Reação em Cadeia da Polimerase Via Transcriptase Reversa/métodos , Transplante Heterólogo , Células Tumorais Cultivadas , Regulação para Cima
8.
Pharmacol Res ; 61(2): 108-15, 2010 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-19643180

RESUMO

DB-67 and its lactone homolog DB-91 are derivatives of topoisomerase I inhibitor camptothecin (CPT) with silyl moiety, which may exhibit a slower inactivation process by changed kinetics of protein binding and/or hydrolysis of its lactone ring and result in increased antitumor activity and decreased toxicity. Pharmacokinetic properties and antitumor activities of the two silatecans were studied and compared. The lactone ring of DB-91 is more stable than those of all the other CPT derivatives in mouse plasma. Both silatecans were metabolized faster than CPT in mouse and human liver microsomes. Pharmacokinetic study revealed a plasma elimination half-life (t(1/2)) of 33 and 94min for DB-67 and DB-91, respectively; similar systemic exposure in plasma between DB-67 and DB-91; and similar volume of distribution at the steady state between DB-67 and DB-91, approximately 15-fold smaller than that of CPT. While DB-91 showed limited activities, DB-67 exhibited activities against the growth of in vivo-like histocultured human tumors and s.c. xenografted human tumors in nude mice. In conclusion, DB-67 is more effective, compared to DB-91, against human tumor growth in in vitro, in vivo-like and in vivo systems. Further pre-clinical and clinical investigations of DB-67 are warranted.


Assuntos
Antineoplásicos Fitogênicos/farmacologia , Camptotecina/análogos & derivados , Inibidores Enzimáticos/farmacologia , Neoplasias/tratamento farmacológico , Compostos de Organossilício/farmacologia , Inibidores da Topoisomerase I , Animais , Antineoplásicos Fitogênicos/administração & dosagem , Antineoplásicos Fitogênicos/sangue , Antineoplásicos Fitogênicos/farmacocinética , Camptotecina/administração & dosagem , Camptotecina/sangue , Camptotecina/farmacocinética , Camptotecina/farmacologia , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Cromatografia Líquida de Alta Pressão , Resistencia a Medicamentos Antineoplásicos , Estabilidade de Medicamentos , Inibidores Enzimáticos/administração & dosagem , Inibidores Enzimáticos/farmacocinética , Feminino , Meia-Vida , Humanos , Injeções Intravenosas , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Nus , Microssomos Hepáticos/metabolismo , Neoplasias/enzimologia , Neoplasias/patologia , Compostos de Organossilício/administração & dosagem , Compostos de Organossilício/sangue , Compostos de Organossilício/farmacocinética , Fatores de Tempo , Carga Tumoral/efeitos dos fármacos , Ensaios Antitumorais Modelo de Xenoenxerto
9.
Mol Cancer ; 8: 120, 2009 Dec 11.
Artigo em Inglês | MEDLINE | ID: mdl-20003326

RESUMO

BACKGROUND: Insulin-like growth factor binding protein (IGFBP-3) is an antiproliferative, pro-apoptotic and invasion suppressor protein which is transcriptionally regulated by p53. Promoter methylation has been linked to gene silencing and cancer progression. We studied the correlation between IGFBP-3 and p53 expression as well as IGFBP-3 promoter methylation in ovarian endometrioid carcinoma (OEC) by immunohistochemical staining and quantitative methylation-specific PCR (qMSP). Additionally, we assessed the molecular regulatory mechanism of wild type (wt) p53 on IGFBP-3 expression using two subclones of OEC, the OVTW59-P0 (low invasive) and P4 (high invasive) sublines. RESULTS: In 60 cases of OEC, 40.0% showed lower IGFBP-3 expression which was significantly correlated with higher IGFBP-3 promoter methylation. p53 overexpression was detected in 35.0% of OEC and was unrelated to clinical outcomes and IGFBP-3. By Kaplan-Meier analysis, patients with lower IGFBP-3, higher IGFBP-3 promoter methylation, and normal p53 were associated most significantly with lower survival rates. In OEC cell line, IGFBP-3 expression was correlated with IGFBP-3 promoter methylation. IGFBP-3 expression was restored after treatment with a DNA methy-transferase inhibitors (5-aza-deoxycytidine) and suppressed by a p53 inhibitor (pifithrin-alpha). The putative p53 regulatory sites on the promoter of IGFBP-3 were identified at -210, -206, -183 and -179 bases upstream of the transcription start site. Directed mutagenesis at these sites quantitatively reduced the transcription activity of IGFBP-3. CONCLUSION: Our data suggests that IGFBP-3 silencing through IGFBP-3 promoter methylation in the absence of p53 overexpression is associated with cancer progression. These results support a potential role of IGFBP-3 methylation in the carcinogenesis of OEC.


Assuntos
Carcinoma Endometrioide/genética , Metilação de DNA , Proteína 3 de Ligação a Fator de Crescimento Semelhante à Insulina/genética , Neoplasias Ovarianas/genética , Regiões Promotoras Genéticas , Proteína Supressora de Tumor p53/genética , Adulto , Feminino , Regulação Neoplásica da Expressão Gênica , Humanos , Imuno-Histoquímica , Pessoa de Meia-Idade , Taxa de Sobrevida , Transcrição Gênica
10.
Ai Zheng ; 28(8): 791-804, 2009 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-19664324

RESUMO

Nasopharyngeal carcinoma (NPC) is one of the common cancers among Chinese living in South China, Taiwan, Singapore, and several other countries or regions in distinct areas. The etiological factors have not been clearly identified yet. So far, no major gene related with hereditary factor has been identified in NPC carcinogenesis; however, some environmental factors, such as consumption of salted fish and long-term exposure to sulfuric-acid vapor, have been tentatively linked to NPC induction, while research has proposed that there is a close association between Epstein-Barr virus (EBV) and NPC pathogenesis. To investigate the relationship between NPC and EBV, we have established ten NPC cell lines. After extensive investigation, we conclude that EBV may establish an infection only in nasopharyngeal neoplastic cells, not in metaplastic epithelial cells, through the IgA receptor (secretory component protein)-mediated endocytosis. Our observations indicate that EBV plays an important role in enhancement of NPC progression, but is involved in neither the initiation nor the promotion of NPC pathogenesis.


Assuntos
Infecções por Vírus Epstein-Barr/virologia , Herpesvirus Humano 4/patogenicidade , Neoplasias Nasofaríngeas/virologia , Receptores Fc/metabolismo , Animais , Linhagem Celular Tumoral , Endocitose , Infecções por Vírus Epstein-Barr/metabolismo , Infecções por Vírus Epstein-Barr/patologia , Infecções por Vírus Epstein-Barr/fisiopatologia , Herpesvirus Humano 4/genética , Herpesvirus Humano 4/isolamento & purificação , Humanos , Queratinas/metabolismo , Neoplasias Nasofaríngeas/metabolismo , Neoplasias Nasofaríngeas/patologia , Neoplasias Nasofaríngeas/fisiopatologia , RNA Viral/metabolismo , Receptores Fc/fisiologia , Sequências Repetidas Terminais , Proteínas da Matriz Viral/metabolismo
11.
J Cell Biochem ; 108(1): 315-25, 2009 Sep 01.
Artigo em Inglês | MEDLINE | ID: mdl-19565568

RESUMO

The identification of molecules that are down-regulated in malignant phenotype is important for understanding tumor biology and their role in tumor suppression. We compared the expression profile of four normal nasal mucosal (NNM) epithelia and a series of nasopharyngeal cancinoma (NPC) cell lines using cDNA microarray and confirmed the actual expression of the selected genes, and found osteoprotegerin (OPG) to be ubiquitously deficient in NPC cells. We also found OPG to be down-regulated in various cancer cell lines, including oral, cervical, ovarian, lung, breast, pancreas, colon, renal, prostate cancer, and hepatoma. Administration of recombinant OPG (rOPG) brought about a reduction in cancer cell growth through apoptotic mechanism. We generated eleven monoclonal antibodies (MAbs) against OPG to study OPG's expression and biological functions in cancer cells. OPG was detected in the tumor stromal regions, but not in the cancer cell per se in surgical specimens of liver cancer. Quantitative reverse transcription-polymerase chain reaction (Q-RT-PCR) revealed that OPG was down-regulated in NPC tissues compared with normal nasal polyp (NNP) tissues. In addition, we showed OPG silencing to be associated with promoter methylation as well as histone modifications. In OPG-silenced cancer cell lines, the OPG gene promoter CpG dinucleotides were highly methylated. Compared to normal cells, silenced OPG gene in cancer cells were found to have reduced histone 3 lysine 4 tri-methylation (H3K4me3) and increased histone 3 lysine 27 tri-methylation (H3K27me3). Taken together, these results suggest that OPG silencing in carcinoma cancer cells occurs through epigenetic repression.


Assuntos
Metilação de DNA , Regulação Neoplásica da Expressão Gênica , Inativação Gênica , Histonas/metabolismo , Neoplasias/genética , Osteoprotegerina/genética , Linhagem Celular Tumoral , Epigênese Genética , Humanos , Masculino , Metilação , Neoplasias/patologia , Osteoprotegerina/metabolismo , Neoplasias da Próstata/genética , Neoplasias da Próstata/metabolismo , Neoplasias da Próstata/patologia
12.
Am J Pathol ; 175(1): 342-54, 2009 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-19541936

RESUMO

Nasopharyngeal carcinoma (NPC) is one of the most common cancers among Chinese living in South China, Singapore, and Taiwan. At present, its etiological factors are not well defined. To identify which genetic alterations might be involved in NPC pathogenesis, we identified genes that were differentially expressed in NPC cell lines and normal nasomucosal cells using subtractive hybridization and microarray analysis. Most NPC cell lines and biopsy specimens were found to have higher expression levels of the gene encoding nucleolar and coiled-body phosphoprotein 1 (NOLC1) as compared with normal cells. Severe combined immunodeficiency mice bearing NPC xenografts derived from NOLC1-short hairpin-RNA-transfected animals were found to have 82% lower levels of tumor growth than control mice as well as marked tumor cell apoptosis. Measuring the expression levels of genes related to cell growth, apoptosis, and angiogenesis, we found that the MDM2 gene was down-regulated in the transfectants. Both co-transfection and chromatin immunoprecipitation experiments showed that tumor protein 53-regulated expression of the MDM2 gene requires co-activation of NOLC1. These findings suggest that NOLC1 plays a role in the regulation of tumorigenesis of NPC and demonstrate that both NOLC1 and tumor protein 53 work together synergistically to activate the MDM2 promoter in NPC cells.


Assuntos
Regulação Neoplásica da Expressão Gênica , Neoplasias Nasofaríngeas/genética , Proteínas Nucleares/genética , Fosfoproteínas/genética , Proteínas Proto-Oncogênicas c-mdm2/biossíntese , Proteína Supressora de Tumor p53/metabolismo , Animais , Western Blotting , Progressão da Doença , Expressão Gênica , Perfilação da Expressão Gênica , Humanos , Imuno-Histoquímica , Imunoprecipitação , Camundongos , Neoplasias Nasofaríngeas/patologia , Análise de Sequência com Séries de Oligonucleotídeos , Interferência de RNA , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Transfecção , Transplante Heterólogo
13.
PLoS One ; 4(1): e4171, 2009.
Artigo em Inglês | MEDLINE | ID: mdl-19137069

RESUMO

Lung cancer is the leading cause of cancer-related mortality worldwide. The lack of tumor specificity remains a major drawback for effective chemotherapies and results in dose-limiting toxicities. However, a ligand-mediated drug delivery system should be able to render chemotherapy more specific to tumor cells and less toxic to normal tissues. In this study, we isolated a novel peptide ligand from a phage-displayed peptide library that bound to non-small cell lung cancer (NSCLC) cell lines. The targeting phage bound to several NSCLC cell lines but not to normal cells. Both the targeting phage and the synthetic peptide recognized the surgical specimens of NSCLC with a positive rate of 75% (27 of 36 specimens). In severe combined immunodeficiency (SCID) mice bearing NSCLC xenografts, the targeting phage specifically bound to tumor masses. The tumor homing ability of the targeting phage was inhibited by the cognate synthetic peptide, but not by a control or a WTY-mutated peptide. When the targeting peptide was coupled to liposomes carrying doxorubicin or vinorelbine, the therapeutic index of the chemotherapeutic agents and the survival rates of mice with human lung cancer xenografts markedly increased. Furthermore, the targeting liposomes increased drug accumulation in tumor tissues by 5.7-fold compared with free drugs and enhanced cancer cell apoptosis resulting from a higher concentration of bioavailable doxorubicin. The current study suggests that this tumor-specific peptide may be used to create chemotherapies specifically targeting tumor cells in the treatment of NSCLC and to design targeted gene transfer vectors or it may be used one in the diagnosis of this malignancy.


Assuntos
Antineoplásicos/administração & dosagem , Carcinoma Pulmonar de Células não Pequenas/tratamento farmacológico , Sistemas de Liberação de Medicamentos/métodos , Neoplasias Pulmonares/tratamento farmacológico , Peptídeos/administração & dosagem , Animais , Antineoplásicos/farmacocinética , Bacteriófagos/imunologia , Linhagem Celular Tumoral , Modelos Animais de Doenças , Doxorrubicina/administração & dosagem , Doxorrubicina/farmacocinética , Humanos , Ligantes , Lipossomos , Camundongos , Camundongos SCID , Biblioteca de Peptídeos , Distribuição Tecidual/efeitos dos fármacos , Vimblastina/administração & dosagem , Vimblastina/análogos & derivados , Vinorelbina
14.
Mol Cancer Ther ; 7(3): 579-89, 2008 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-18347144

RESUMO

Hepatocellular carcinoma is the fourth leading cause of cancer death worldwide. Novel treatment strategies derived from increased knowledge of molecular oncology are constantly being developed to cure this disease. Here, we used phage display to identify a novel peptide (SP94), which binds specifically to hepatocellular carcinoma cells. In vitro, the phage clone PC94 was shown to bind to hepatocellular carcinoma cell lines by ELISA and flow cytometry analysis. In vivo, PC94 homed specifically to tumor tissues but not to normal visceral organs in severe combined immunodeficient mice bearing human hepatocellular carcinoma xenografts. This homing ability could be competitively inhibited by synthetic peptide, SP94. Immunohistochemical staining confirmed that PC94 localized to tumor tissues and that it could not be detected in SP94-competed tumor tissues. In addition, PC94 recognized the tumor tissue but not nontumor tissue in surgical specimens from hepatocellular carcinoma patients, with a positive rate of 61.3% (19 of 31). With the conjugation of SP94 and liposomal doxorubicin, the targeted drug delivery system enhanced the therapeutic efficacy against hepatocellular carcinoma xenografts through enhanced tumor apoptosis and decreased tumor angiogenesis. Our results indicate that SP94 has the potential to improve the systemic treatment of patients with advanced hepatocellular carcinoma.


Assuntos
Carcinoma Hepatocelular/tratamento farmacológico , Sistemas de Liberação de Medicamentos , Neoplasias Hepáticas/tratamento farmacológico , Peptídeos/administração & dosagem , Animais , Linhagem Celular Tumoral , Ensaio de Imunoadsorção Enzimática , Citometria de Fluxo , Humanos , Ligantes , Camundongos , Camundongos SCID
15.
Cancer Res ; 67(22): 10958-65, 2007 Nov 15.
Artigo em Inglês | MEDLINE | ID: mdl-18006841

RESUMO

Antiangiogenesis therapies for the treatment of cancers hold the promise of high efficacy and low toxicity. In vivo phage display was used to identify peptides specifically targeting tumor blood vessels. The peptide SP5-52 recognized tumor neovasculature but not normal blood vessels in severe combined immunodeficiency mice bearing human tumors. Synthetic peptide was shown to inhibit the binding of PC5-52 phage particles to the tumor mass in the competitive inhibition assay. Several selected phage clones displayed the consensus motif, proline-serine-proline, and this motif was crucial for peptide binding to the tumor neovasculature. SP5-52 peptides also bound vascular endothelial growth factor-stimulated human umbilical vein endothelial cells and blood vessels of human lung cancer surgical specimens. Furthermore, this targeting phage was shown to home to tumor tissues from eight different types of human tumor xenografts following in vivo phage display experiments. An SP5-52 peptide-linked liposome carrying doxorubicin enhanced the therapeutic efficacy of the drug, markedly decreased tumor blood vessels, and resulted in higher survival rates of human lung and oral cancer-bearing xenograft mice. The current study indicates that ligand-targeted therapy offers improved therapeutic effects over conventional anticancer drug therapy, and that the peptide SP5-52 specifically targets tumor neovasculature and is a good candidate for targeted drug delivery to solid tumors.


Assuntos
Sistemas de Liberação de Medicamentos , Neoplasias Pulmonares/tratamento farmacológico , Neoplasias Pulmonares/patologia , Neovascularização Patológica , Peptídeos/administração & dosagem , Peptídeos/química , Animais , Linhagem Celular Tumoral , Modelos Animais de Doenças , Doxorrubicina/administração & dosagem , Doxorrubicina/farmacologia , Endotélio Vascular/citologia , Humanos , Camundongos , Camundongos SCID , Neoplasias Bucais/tratamento farmacológico , Transplante de Neoplasias , Biblioteca de Peptídeos
16.
Clin Vaccine Immunol ; 14(4): 404-11, 2007 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-17287314

RESUMO

Dengue virus (DEN), the pathogen behind dengue hemorrhagic fever, remains a public health problem in Asia and South America. In this study, monoclonal antibodies (MAbs) against DEN serotype 1 (DEN-1) were generated by fusing NSI/1-Ag4-1 mouse myeloma cells with lymphocytes from BALB/c mice immunized with DEN-1. Twelve MAbs were found to react specifically to the DENs by enzyme-linked immunosorbent assay, immunofluorescence analysis, and immunoblotting analysis. Five MAbs, namely, DA4-7, DA6-7, DA9-5, DA10-2, and DA11-13, were found to react with envelope proteins of DEN-1. Two serotype-specific MAbs of DEN-1, DA6-7 and DA11-13, were further shown to neutralize DEN-1 infection by a plaque reduction neutralization test. The neutralizing epitopes of these MAbs were further identified from a random peptide library displayed on phage. Immunopositive phage clones reacted specifically with these MAbs and did not react with normal mouse serum. Epitope-based peptide antigens were proved able to detect antibodies in serum samples collected from DEN-1-infected patients but not in those taken from DEN-2-infected patients or healthy controls. We believe that these MAbs and neutralizing epitopes will provide information that will lead to the development of DEN-1 serotype-specific diagnostic reagents and vaccines.


Assuntos
Anticorpos Monoclonais/biossíntese , Antígenos Virais/imunologia , Vírus da Dengue/imunologia , Mapeamento de Epitopos , Peptídeos/imunologia , Animais , Anticorpos Monoclonais/sangue , Anticorpos Monoclonais/química , Antígenos Virais/sangue , Linhagem Celular , Humanos , Camundongos , Peptídeos/sangue
17.
Funct Integr Genomics ; 7(1): 79-93, 2007 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-16988809

RESUMO

It was proposed that Epstein-Barr virus (EBV) is closely associated with nasopharyngeal carcinoma (NPC); however, the molecular mechanisms involved in the effect of EBV on NPC host genes have not yet been well defined. For this study, two sets of microarray experiments, NPC (EBV-free) vs normal epithelial cells and EBV(+) vs EBV(-) NPC arrays, were analyzed and the datasets were cross-compared to identify any correlation between gene clusters involved in EBV targeting and the NPC host gene expression profiles. Statistical analysis revealed that EBV seems to have a preference for targeting more genes from the differentially expressed group in NPC cells than those from the ubiquitously expressed group. Furthermore, this trend is also reflected in log ratios where the EBV target genes of the differentially expressed group origin showed greater log ratios than genes with an origin from the ubiquitously expressed NPC group. Taken together, the genome-wide comparative scanning of EBV and NPC transcriptomes has successfully demonstrated that EBV infection has an intensifying effect on the signals involved in NPC gene expression both in breadth (the majority of the genes) and in depth (greater log ratios).


Assuntos
Carcinoma/virologia , Infecções por Vírus Epstein-Barr/genética , Regulação Neoplásica da Expressão Gênica/fisiologia , Herpesvirus Humano 4/fisiologia , Neoplasias Nasofaríngeas/virologia , Carcinoma/genética , Carcinoma/metabolismo , Linhagem Celular , Células Cultivadas , Infecções por Vírus Epstein-Barr/metabolismo , Humanos , Neoplasias Nasofaríngeas/genética , Neoplasias Nasofaríngeas/metabolismo
18.
Planta Med ; 71(1): 28-32, 2005 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-15678370

RESUMO

The mechanism of action, leakage of cytochrome c from mitochondria into cytosol, for the antineoplastic compound glyfoline was examined. Additionally, our current studies revealed that glyfoline induced apoptotic changes and arrested cell cycle procession at the G2/M phase in nasopharyngeal carcinoma (NPC). A reverse transcriptase polymerase chain reaction (RT-PCR) showed no specific changes of apoptosis-related gene expression (i. e., bax, ICE-alpha,beta, bcl-2, and c-myc). However, no similar changes were detected in fibroblasts and peripheral lymphocytes after glyfoline treatment suggesting that glyfoline has a higher affinity for tumor cells than for normal cells.


Assuntos
Acridinas/farmacologia , Antineoplásicos Fitogênicos/farmacologia , Citocromos c/efeitos dos fármacos , Fitoterapia , Rutaceae , Acridinas/administração & dosagem , Acridinas/uso terapêutico , Antineoplásicos Fitogênicos/administração & dosagem , Antineoplásicos Fitogênicos/uso terapêutico , Apoptose/efeitos dos fármacos , Apoptose/genética , Linhagem Celular Tumoral/efeitos dos fármacos , Linhagem Celular Tumoral/metabolismo , Citocromos c/metabolismo , DNA de Neoplasias/análise , Regulação Neoplásica da Expressão Gênica , Humanos , Concentração Inibidora 50 , Mitocôndrias/efeitos dos fármacos , Mitocôndrias/metabolismo , Neoplasias Nasofaríngeas/genética , Neoplasias Nasofaríngeas/patologia , Extratos Vegetais/administração & dosagem , Extratos Vegetais/farmacologia , Extratos Vegetais/uso terapêutico , Reação em Cadeia da Polimerase Via Transcriptase Reversa
19.
Cancer Res ; 64(21): 8002-8, 2004 Nov 01.
Artigo em Inglês | MEDLINE | ID: mdl-15520208

RESUMO

Nasopharyngeal carcinoma (NPC) is a common cancer among Chinese living in southern China, Taiwan, and Singapore. The 5-year survival rate in the early stage of NPC has been reported as high as 90 to 95% with the use of radiotherapy, but in the advanced cases, even with the use of both chemotherapy and radiotherapy, the survival rate is still <50%. To improve the survival rate, we identify a 12-mer peptide (L-peptide) specifically binding to NPC cells with a phage displayed random peptide library. The L-phage and synthetic L-peptide bound to the tumor cell surfaces of most NPC cell lines and biopsy specimens, but not normal nasal mucosal cells, and the L-peptide-linked liposomes containing fluorescent substance (L-peptide-Lipo-HPTS) were capable of binding to and translocating across plasma membranes. L-Peptide-linked liposomes that carried doxorubicin (L-peptide-Lipo-Dox) caused marked cytotoxicity in NPC cells. In SCID mice bearing NPC xenografts, the L-phages specifically bound to the tumor mass, an effect that was inhibited by competition with synthetic L-peptide. In addition, the L-peptide-Lipo-Dox suppressed tumor growth better than Lipo-Dox. These results indicate that the novel L-peptide specifically binds NPC cells and is a good candidate for targeted drug delivery to NPC solid tumors.


Assuntos
Doxorrubicina/administração & dosagem , Neoplasias Nasofaríngeas/tratamento farmacológico , Peptídeos/administração & dosagem , Animais , Sítios de Ligação , Endocitose , Lipossomos , Camundongos , Camundongos SCID , Transplante de Neoplasias , Biblioteca de Peptídeos , Peptídeos/metabolismo , Transplante Heterólogo
20.
Lab Invest ; 84(12): 1547-56, 2004 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-15448710

RESUMO

To understand whether the p53-regulated mdm2 gene expression was altered by the Epstein-Barr virus (EBV) in nasopharyngeal carcinoma (NPC), the NPC-TW01 cell line was infected by EBV through IgA receptor-mediated endocytosis. The mdm2 gene was expressed only in a small fraction of the NPC cell population and could be enhanced in the EBV-infected (EBV+) cells. In the animals bearing EBV+ and EBV- NPC xenografts, the MDM2+ cells only appeared in clusters in both EBV+ and EBV- tumors with stronger expression in EBV+ cells. Cotransfection of pmdm2-Luc plus pSV40-p53 plus pCMV-LMP1 in the NPC-TW06 line that had p53 heterozygous point mutation showed stronger mdm2 promoter activity than cells cotransfected with pmdm2-Luc plus pSV40-p53, but no mdm2 promoter activity was seen in cells cotransfected with pmdm2-Luc plus pCMV-LMP1. Only the EBV-LMP1 but not the EBV-LMP2A gene could enhance p53 to upregulated mdm2 expression. Tumor cells in NPC biopsy specimens revealed similar mdm2 expression as in the animal model. It is concluded that although EBV can indirectly enhance mdm2 gene expression in tumor cells that express this gene, it cannot turn on or directly regulate mdm2 expression in cells that do not express this gene. In other words, EBV plays a role as an enhancer in NPC tumorigenesis.


Assuntos
Regulação Neoplásica da Expressão Gênica/genética , Regulação Viral da Expressão Gênica/genética , Herpesvirus Humano 4/genética , Neoplasias Nasofaríngeas/genética , Proteínas Nucleares/genética , Proteínas Proto-Oncogênicas/genética , Animais , Carcinoma de Células Escamosas/genética , Linhagem Celular Tumoral , Endocitose , Camundongos , Camundongos SCID , Neoplasias Nasofaríngeas/virologia , Proteínas Proto-Oncogênicas c-mdm2 , Transplante Heterólogo , Dedos de Zinco
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