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1.
World J Gastrointest Oncol ; 16(3): 979-990, 2024 Mar 15.
Artigo em Inglês | MEDLINE | ID: mdl-38577474

RESUMO

BACKGROUND: Helicobacter pylori (H. pylori) is the primary risk factor for gastric cancer (GC), the Wnt/ß-Catenin signaling pathway is closely linked to tumourigenesis. GC has a high mortality rate and treatment cost, and there are no drugs to prevent the progression of gastric precancerous lesions to GC. Therefore, it is necessary to find a novel drug that is inexpensive and preventive to against GC. AIM: To explore the effects of H. pylori and Moluodan on the Wnt/ß-Catenin signaling pathway and precancerous lesions of GC (PLGC). METHODS: Mice were divided into the control, N-methyl-N-nitrosourea (MNU), H. pylori + MNU, and Moluodan groups. We first created an H. pylori infection model in the H. pylori + MNU and Moluodan groups. A PLGC model was created in the remaining three groups except for the control group. Moluodan was fed to mice in the Moloudan group ad libitum. The general condition of mice were observed during the whole experiment period. Gastric tissues of mice were grossly and microscopically examined. Through quantitative real-time PCR (qRT-PCR) and Western blotting analysis, the expression of relevant genes were detected. RESULTS: Mice in the H. pylori + MNU group showed the worst performance in general condition, gastric tissue visual and microscopic observation, followed by the MNU group, Moluodan group and the control group. QRT-PCR and Western blotting analysis were used to detect the expression of relevant genes, the results showed that the H. pylori + MNU group had the highest expression, followed by the MNU group, Moluodan group and the control group. CONCLUSION: H. pylori can activate the Wnt/ß-catenin signaling pathway, thereby facilitating the development and progression of PLGC. Moluodan suppressed the activation of the Wnt/ß-catenin signaling pathway, thereby decreasing the progression of PLGC.

2.
Tumori ; 107(6): 504-513, 2021 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-33272103

RESUMO

OBJECTIVE: To investigate the mechanism of long noncoding RNA (lncRNA) DANCR on the progression of hepatocellular carcinoma (HCC) cells. METHODS: The expression levels of DANCR and miR-125b-5p were measured in normal hepatocytes (LO2) and HCC cell lines by quantitative reverse transcription polymerase chain reaction. HepG2 and Huh-7 cells were transfected with sh-DANCR, the negative control (sh-NC), miR-125b-5p mimic, or mimic NC or cotransfected with sh-DANCR and miR-125b-5p inhibitor. HCC cell proliferation was assessed through CCK8 and plate colony formation assay. Western blot quantified the expression levels of Bcl-2, Bax, caspase-3, and cleaved-caspase-3. Apoptotic rate was detected as well as migratory and invasive capacities. The implication of the MAPK signal pathway was assessed by detecting the expression levels of p38, ERK1/2, JNK, p-p38, p-ERK1/2, and p-JNK. Interactions between DANCR and miR-125b-5p were detected by dual luciferase reporter assay. RESULTS: In HCC cells, DANCR was highly expressed and miR-125b-5p was decreased. sh-DANCR or miR-125b-5p mimic stimulation reduced HepG2 or Huh-7 cell progression while promoted cell apoptosis evidenced by increased apoptotic rate, elevated levels of Bax and cleaved-caspase-3, and decreased Bcl-2. Moreover, the migration rate and invasiveness of HCC cells were also inhibited by sh-DANCR and miR-125b-5p mimic. Levels of p-p38/p38, p-ERK1/2/ERK1/2, and p-JNK/JNK were suppressed by sh-DANCR and miR-125b-5p mimic. LncRNA DANCR negatively targeted and directly bound to miR-125b-5p. Knockdown of miR-125b-5p could reverse the inhibitory effects of sh-DANCR on HCC cells. CONCLUSION: In HCC cells, lncRNA DANCR sponges miR-125b-5p and activates MAPK pathway, thus facilitating HCC cell progression.


Assuntos
Carcinoma Hepatocelular/genética , Regulação Neoplásica da Expressão Gênica , Neoplasias Hepáticas/genética , MicroRNAs/genética , Interferência de RNA , RNA Longo não Codificante/genética , Regiões 3' não Traduzidas , Apoptose/genética , Biomarcadores Tumorais , Carcinoma Hepatocelular/metabolismo , Carcinoma Hepatocelular/patologia , Linhagem Celular Tumoral , Movimento Celular/genética , Proliferação de Células/genética , Citometria de Fluxo , Genes Reporter , Humanos , Neoplasias Hepáticas/metabolismo , Neoplasias Hepáticas/patologia , Sistema de Sinalização das MAP Quinases
3.
BMC Cancer ; 19(1): 1219, 2019 Dec 16.
Artigo em Inglês | MEDLINE | ID: mdl-31842811

RESUMO

BACKGROUND: The posterior-inferior border of symphysis (PIBS) point system is a novel vaginal dose-reporting method and is a simple and reliable method proposed by the Medical University of Vienna proposed for both external-beam radiotherapy (EBRT) and brachytherapy (BT). In this multicenter study, we sought to first evaluate the vaginal radiation dose in Chinese cervical cancer patients according to the PIBS point system and then to analyze the factors influencing the dose distribution. METHODS: We collected data from the medical records of 936 cervical cancer patients who underwent concurrent radiochemotherapy at 13 different institutions in China. Radiation doses at points A, PIBS+ 2 cm, PIBS and PIBS-2 cm, International Commission on Radiation Units (ICRU)-R and ICRU-B were measured. RESULTS: The median total doses in EQD2α/ß = 3 at points PIBS+ 2 cm, PIBS and PIBS-2 cm were 82.5 (52.7-392.1) Gy, 56.2 (51.4-82.1) Gy and 2.6 (0.9-7.4) Gy, respectively. The median total doses in EQD2α/ß = 3 at ICRU-R and ICRU-B were 77.5 (54.8-132.4) Gy and 79.9 (60.7-133.7) Gy, respectively. The mean vaginal reference length (VRL) was 4.6 ± 1.0 cm (median, 4.5 cm). In patients with VRL ≤4.5 cm, the mean total doses in EQD2α/ß = 3 at points PIBS+ 2 cm, PIBS and PIBS-2 cm were 128.5, 60.7 and 0.8 Gy, respectively. In patients with VRL > 4.5 cm, the mean total doses at these three points were 68.9, 0.5 and 54.5 Gy, respectively. Classification of patients revealed significant differences (P < 0.05) between these two groups. CONCLUSIONS: With the PIBS point system, Chinese patients with a shorter VRL of < 4.5 cm received higher radiation doses at the PIBS+ 2 cm, PIBS and PIBS-2 cm points than European and American patients. Further studies are required to establish the dose-effect relationships with these points as references. The study was registered as a clinical trial (NCT03257475) on August 22, 2017.


Assuntos
Braquiterapia , Quimiorradioterapia , Neoplasias do Colo do Útero/terapia , Adenocarcinoma/terapia , Adulto , Povo Asiático , Carcinoma de Células Escamosas/terapia , China , Feminino , Humanos , Pessoa de Meia-Idade , Dosagem Radioterapêutica
4.
Chem Biodivers ; 16(12): e1900471, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31612620

RESUMO

One natural p-terphenyl glycoside, gliocladinin C, and two furano-polyene derivatives, chaetominins A and B, were isolated from potato endophytic fungus Chaetomium subaffine. The absolute configurations of these compounds were elucidated by HR-ESI-MS, NMR, the DP4+ probabilities and electronic circular dichroism (ECD) spectra. Furthermore, gliocladinin C and chaetominin A showed cytotoxic activity against two selected human tumor cell lines (Hep-2 and HepG-2).


Assuntos
Antineoplásicos/química , Chaetomium/metabolismo , Compostos de Terfenil/química , Antineoplásicos/farmacologia , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos dos fármacos , Chaetomium/química , Dicroísmo Circular , Humanos , Espectroscopia de Ressonância Magnética , Conformação Molecular , Compostos de Terfenil/farmacologia
5.
Immunol Invest ; 48(8): 809-821, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31405308

RESUMO

Aims The aryl hydrocarbon receptor (AhR) plays a pivotal role in regulating the innate and the acquired immune systems. The present study aimed to investigate the association of Crohn's disease (CD) with AhR polymorphisms in a cohort of patients from Southeast China. Methods An improved multiple ligase detection reaction technique was applied to examine the polymorphisms of rs2158041, rs2066853, and rs10249788 in 310 patients with CD and 573 controls. Results Compared to the controls, the variant allele (T) and genotype (CT+TT) of rs2158041 were less frequent in patients with CD (both p < 0.05). Similar conclusions were drawn from patients with ileal CD and with stricture CD as compared to the controls (all p < 0.0083). However, no significant differences were observed in allele and genotype frequencies of rs2066853 and rs10249788 between patients with CD and the controls (all p > 0.05). Although rs2158041 and rs10249788 were in complete linkage disequilibrium with rs2066853, respectively, only the frequency of haplotype (TG) formed by rs2158041 and rs2066853 was significantly lower in patients with CD than that in the controls (p < 0.05). Conclusions AhR (rs2158041) might be a susceptible locus for CD, especially for the two subtypes: ileal CD and stricture CD.


Assuntos
Fatores de Transcrição Hélice-Alça-Hélice Básicos/genética , Doença de Crohn/genética , Predisposição Genética para Doença/genética , Polimorfismo de Nucleotídeo Único , Receptores de Hidrocarboneto Arílico/genética , Adulto , Alelos , Povo Asiático/genética , China , Doença de Crohn/etnologia , Feminino , Frequência do Gene , Predisposição Genética para Doença/etnologia , Genótipo , Haplótipos , Humanos , Desequilíbrio de Ligação , Masculino , Pessoa de Meia-Idade , Adulto Jovem
6.
Cell Physiol Biochem ; 33(1): 97-106, 2014.
Artigo em Inglês | MEDLINE | ID: mdl-24480980

RESUMO

BACKGROUND & AIMS: MicroRNAs (miRNAs) have been shown to play essential roles in HSCs activation which contributes to hepatic fibrosis. Our previous miRNA microarray results suggested that miR-126 might be decreased during HSCs activation as other studies. The aim of this study is to investigate the role of miR-126 during HSCs activation. METHODS: In this study, the expression of miR-126 during HSCs activation was measured and confirmed by qRT-PCR. Then, miR-126 expression was restored by transfection of lentivirus vector encoding miR-126. Futhermore, cell proliferation was assayed by the cell counting kit-8 (CCK-8), cell migration was assayed by transwell assay, and the markers of activation of HSCs, α-SMA and collagen type I, were assayed by qRT-PCR, Western Blotting, Immunostaining and ELISA. Luciferase reporter assay was used to find the target of miR-126, and Western Blotting and Immunostaining was used to validate the target of miR-126. Then, the expression and the role of the target of miR-126 during HSCs activation was further assessed. RESULTS: The expression of miR-126 was confirmed to be significantly decreased during HSCs activation. Overexpression of miR-126 significantly inhibited HSCs migration but did not affect HSCs proliferation. The expression of α-SMA and collagen type I were both obviously decreased by miR-126 restoration. CRK was found to be the target of miR-126 and overexpression of miR-126 significantly inhibited CRK expression. And it was found that overexpression of CRK also significantly decreased miR-126 expression and promoted HSCs activation. CONCLUSIONS: Our study showed that overexpression of miR-126 significantly inhibited the activation and migration of HSCs through targeting CRK which can also decrease miR-126 expression and promote HSCs activation.


Assuntos
Movimento Celular , Células Estreladas do Fígado/citologia , Células Estreladas do Fígado/metabolismo , MicroRNAs/metabolismo , Proteínas Proto-Oncogênicas c-crk/metabolismo , Animais , Sequência de Bases , Linhagem Celular , Proliferação de Células , Colágeno Tipo I/metabolismo , Regulação da Expressão Gênica , Masculino , MicroRNAs/genética , Dados de Sequência Molecular , Ratos Sprague-Dawley
7.
Zhonghua Liu Xing Bing Xue Za Zhi ; 34(7): 668-72, 2013 Jul.
Artigo em Chinês | MEDLINE | ID: mdl-24257165

RESUMO

OBJECTIVE: To explore the setting of logos on tobacco control information at outlets for retails and restaurants in 12 selected cities of China. METHODS: For all the shops for retail of tobacco, alcohol, food and restaurants under survey in 333 blocks of 12 cities(Beijing, Tianjin, Shanghai, Qingdao, Hangzhou, Shaoxing, Suzhou, Nantong, Zhenjiang, Chengdu, Xining and Harbin), setting and contents of logos on tobacco control information, inside and outside them were examined. RESULTS: 45 700 objectives were included in the study. Among all types of retail shops, the identification rate of tobacco control information at the entrance and inside were 3.6% and 4.4%, with an overall identification rate as 7.0%. The overall rate at the entrance of all the restaurants was 4.6% which was larger than the ones at the retail shops. Our result showed that there were differences between cities and types of establishments and higher rates seen in the larger ones. Of all the places that having had placement of information on tobacco control, only 18.5% of them had put them both inside and outside. Slogans or images on "No Smoking" were the main forms of information but less than 10% of them would show signs as 'exclusive non-smoking'. CONCLUSION: Data from our survey showed that the identification rate of tobacco control information was at a low level in 12 cities, and differences were seen between cities, size of establishment, that called for improvement of the existing tobacco control policies in China.


Assuntos
Diretórios de Sinalização e Localização/estatística & dados numéricos , Saúde Pública , Prevenção do Hábito de Fumar , China , Cidades , Restaurantes
8.
Cell Biol Int ; 33(4): 509-15, 2009 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-19353779

RESUMO

Gene-directed enzyme prodrug therapy (GDEPT) is a strategy developed to selectively target cancer cells. However, the clinical benefit is limited due to its poor gene transfer efficiency. To overcome this obstacle, we took advantage of piggyBac (PB) transposon, a natural non-viral gene vector that can induce stable chromosomal integration and persistent gene expression in vertebrate cells, including human cells. To determine whether the vector can also mediate stable gene expression in ovarian cancer cells, we constructed a PB transposon system that simultaneously expresses the Herpes simplex virus thymidine kinase (HSV-tk) gene and the monomeric red fluorescent protein (mRFP1) reporter gene. The recombinant plasmid, pPB/TK, was transfected into ovarian adenocarcinoma cells SKOV3 with FuGENE HD reagent, and the efficiency was given by the percentage of mRFP1-positive cells detected by flow cytometry and confocal microscopy. The specific expression of HSV-tk in transfected cells was confirmed by RT-PCR and western blotting. The sensitivity of transfected cells to pro-drug ganciclovir (GCV) was determined by methylthiazoletetrazolium (MTT) assay. A total of 56.4 +/- 8.4% cells transfected with pPB/TK were mRFP1 positive, compared to no measurable mRFP1 expression in pORF-HSVtk-transfected cells. The expression level of HSV-tk in pPB/TK-transfected cells was 10 times higher than in pORF-HSVtk-transfected cells. The results show that pPB/TK transfection increases the sensitivity of cells to GCV in a dose-dependent manner. Our data indicate that the PB transposon system could enhance the anti-tumor efficiency of GDEPT in ovarian cancer.


Assuntos
Adenocarcinoma/terapia , Elementos de DNA Transponíveis/genética , Terapia Genética/métodos , Vetores Genéticos/genética , Neoplasias Ovarianas/terapia , Pró-Fármacos/farmacologia , Linhagem Celular Tumoral , Resistencia a Medicamentos Antineoplásicos , Feminino , Ganciclovir/farmacologia , Humanos , Proteínas Luminescentes/genética , Proteínas Recombinantes de Fusão/genética , Proteínas Recombinantes de Fusão/metabolismo , Timidina Quinase/genética , Transfecção , Proteína Vermelha Fluorescente
9.
Cell Res ; 16(1): 82-92, 2006 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-16467879

RESUMO

N-acetylglucosaminyltransferase V (GnT-V) is an important tumorigenesis and metastasis-associated enzyme. To study its biofunction, the GnT-V stably suppressed cell line (GnT-V-AS/7,721) was constructed from 7,721 hepatocarcinoma cells in previous study. In this study, cDNA array gene expression profiles were compared between GnT-V-AS/7,721 and parental 7,721 cells. The data indicated that GnT-V-AS/7,721 showed a characteristic expression pattern consistent with the ER stress. The molecular mechanism of the ER stress was explored in GnT-V-AS/7,721 by the analysis on key molecules in both two unfolded protein response (UPR) pathways. For ATF6 and Ire1/XBP-1 pathway, it was evidenced by the up-regulation of BIP at mRNA and protein level, and the appearance of the spliced form of XBP-1. As for PERK/eIF2alpha pathway, the activation of ER eIF2alpha kinase PERK was observed. To confirm the results from GnT-V-AS/7,721 cells, the key molecules in the UPR were examined again in 7,721 cells interfered with the GnT-V by the specific RNAi treatment. The results were similar with those from GnT-V-AS/7721, indicating that blocking of GnT-V can specifically activate ER stress in 7,721 cells. Rate of (3)H-Man incorporation corrected with rate of (3)H-Leu incorporation in GnT-V-AS/7,721 was down-regulated greatly compared with the control, which demonstrated the deficient function of the enzyme synthesizing N-glycans after GnT-V blocking. Moreover, the faster migrating form of chaperone GRP94 associated with the underglycosylation, and the extensively changed N-glycans structures of intracellular glycoproteins were also detected in GnT-V-AS/7,721. These results supported the mechanism that blocking of GnT-V expression impaired functions of chaperones and N-glycan-synthesizing enzymes, which caused UPR in vivo.


Assuntos
Carcinoma Hepatocelular/metabolismo , Retículo Endoplasmático/fisiologia , Regulação Neoplásica da Expressão Gênica , N-Acetilglucosaminiltransferases/metabolismo , Linhagem Celular Tumoral , Proteínas de Ligação a DNA/metabolismo , Perfilação da Expressão Gênica , Humanos , Proteínas Reguladoras de Ferro/metabolismo , Glicoproteínas de Membrana/metabolismo , N-Acetilglucosaminiltransferases/fisiologia , Análise de Sequência com Séries de Oligonucleotídeos , Polissacarídeos/biossíntese , Polissacarídeos/química , Dobramento de Proteína , Interferência de RNA , Fatores de Transcrição de Fator Regulador X , Transdução de Sinais , Fatores de Transcrição/metabolismo , Transfecção , Proteína 1 de Ligação a X-Box , eIF-2 Quinase/metabolismo
10.
Vaccine ; 24(12): 2141-50, 2006 Mar 15.
Artigo em Inglês | MEDLINE | ID: mdl-16368168

RESUMO

Fusing dendritic cells (DCs) with tumor cells is a powerful vaccine to increase tumor immunogenicity. To develop more effective and safer therapeutic vaccine, we fused rat bone marrow-derived DCs with ovarian tumor cell line NuTu-19 modified by suicide gene (HSV1-TK gene) to obtain live vaccine against ovarian cancer. Our data showed that immunization of rats with such live vaccine solicited stronger ovarian tumor-specific cytotoxic T lymphocyte responses and induced immunopreventive and immunotherapeutic effects against parental tumor cells in vivo. Live vaccine could be induced to death after ganciclovir administration in vitro and in vivo. Our researches suggest that live vaccine modified with suicide gene might be effective and controllable in the therapy of ovarian cancer.


Assuntos
Vacinas Anticâncer/imunologia , Genes Transgênicos Suicidas , Terapia Genética/métodos , Herpesvirus Humano 1/metabolismo , Neoplasias Ovarianas/terapia , Animais , Apoptose/fisiologia , Vacinas Anticâncer/administração & dosagem , Vacinas Anticâncer/genética , Fusão Celular , Linhagem Celular Tumoral , Células Dendríticas/imunologia , Feminino , Herpesvirus Humano 1/genética , Neoplasias Ovarianas/imunologia , Ratos , Timidina Quinase/genética , Timidina Quinase/metabolismo
11.
DNA Seq ; 16(4): 295-9, 2005 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-16147889

RESUMO

Human THROMBOSPONDIN-1 play versatile roles in platelet aggregation, angiogenesis, and tumorigenesis, which forms a disulfide-linked homotrimeric complex. Here we reported that the genomic lotus of TSP1 also transcribes a natural antisense transcript (NAT) with an overlapping and reverse complement region against TSP1. The NAT of TSP1 was expressed in human testis, lung, thymus, colon, placenta, kidney and skeleton muscle, revealed by PCR amplification. It was also expressed in some tumor cell lines. The identification of NAT of TSP1 would be of significant importance to understand the functions of TSP1, and it would also suggest the potential attempt of using RNA interference for related tumor therapy, for instance, breast cancer.


Assuntos
DNA Antissenso/genética , Especificidade da Espécie , Trombospondina 1/genética , Sequência de Bases , Linhagem Celular Tumoral , DNA Antissenso/metabolismo , Expressão Gênica , Humanos , Dados de Sequência Molecular , Trombospondina 1/metabolismo , Distribuição Tecidual
12.
Ai Zheng ; 24(8): 909-14, 2005 Aug.
Artigo em Chinês | MEDLINE | ID: mdl-16086864

RESUMO

BACKGROUND & OBJECTIVE: Live tumor vaccines could achieve better immunotherapeutic effects than irradiated tumor vaccines; however, the tumorigenicity is the crucial drawback incurred by the current procedures for vaccine preparation. This study was to explore the application of herpes simplex virus type 1 thymidine kinase/ganciclovir (HSV1-TK/GCV) system as "in vivo death switch" to control the survival status of cancer vaccines under certain circumstances. METHODS: Suicide gene HSV(1)-TK was transferred into ovarian cancer cell line NuTu-19 with a retrovirus vector, followed by G418 selection to obtain HSV(1)-TK-transfected NuTu-19 cells (NuTu-19/TK). Dendritic cells (DCs) derived from Fischer344 rat bone marrow were fused with NuTu-19/TK cells to construct live tumor vaccine FC/TK. The expression of HSV(1)-TK in FC/TK cells was determined by reverse transcription-polymerase chain reaction (RT-PCR) and Western blot. The cytotoxic efficacy of GCV on FC/TK cells was evaluated by XTT assay. The apoptosis of FC/TK cells was analyzed by flow cytometry and Hoechst33258 staining after GCV treatment. Rats were vaccinated with FC/TK cells and divided into 2 groups: GCV group (5 rats) were intraperitoneally treated with GCV for 7 days, control group (5 rats) were treated with normal saline. The tumorigenesis and tumor metastasis in the rats were observed 90 days after inoculation. RESULTS: HSV1-TK was specifically expressed in FC/TK cells. GCV showed in vitro cytotoxicity to FC/TK cells in a dose-dependent manner, and 86.25% of the FC/TK cells were killed by GCV at a concentration of 100 microg/ml; the apoptosis rate of FC/TK cells was over 80%, and apoptotic morphology, including cell shrinkage, chromatin condensation, was observed. In the rat models, the tumor was developed at the injection site in 3 rats of control group, while no tumor was observed in the rats of GCV group. CONCLUSION: HSV(1)-TK/GCV could act as the "death switch" of tumor vaccines by triggering apoptosis of tumor vaccines both in vitro and in vivo.


Assuntos
Vacinas Anticâncer , Ganciclovir/farmacologia , Genes Transgênicos Suicidas , Herpesvirus Humano 1/enzimologia , Neoplasias Ovarianas/patologia , Timidina Quinase/metabolismo , Animais , Apoptose/efeitos dos fármacos , Fusão Celular , Células Dendríticas/citologia , Células Dendríticas/enzimologia , Feminino , Vetores Genéticos , Herpesvirus Humano 1/genética , Imunoterapia Ativa , Neoplasias Ovarianas/enzimologia , Ratos , Ratos Endogâmicos F344 , Retroviridae/genética , Timidina Quinase/genética , Transfecção , Células Tumorais Cultivadas
13.
J Cancer Res Clin Oncol ; 131(3): 157-62, 2005 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-15599595

RESUMO

PURPOSE: Bladder carcinoma is the most common urological malignancy in China. Gene mutation may be one of causes of carcinogenesis in the cancer. We therefore investigated the mRNA expression of RTKN gene in clinic malignant bladder carcinoma and explored the relationship between the novel gene and the cancer. METHODS: Total RNA was extracted from 33 surgically resected specimens of bladder carcinoma and 19 specimens of tumor-free bladder tissues. After the optimal reverse-transcription polymerase chain reaction condition was established, the mRNA expression levels of the RTKN gene in the lesions and tumor-free bladder tissues were examined semiquantitatively, and the relationships between expression levels of RTKN and clinical pathological features were analyzed. RESULTS: The expression of RTKN gene mRNA in 33 human bladder carcinoma tissues was significantly higher than that in 19 human tumor-free bladder tissues (0.937+/-0.103 vs. 0.350+/-0.082). The average ratio of RTKN expression in neoplasms to that in tumor-free bladder tissues was 0.350+/-0.164. Based on this ratio the 33 patients were divided into three groups: a down-regulated expression group (n=2), an up-regulated expression group (n=22), and an unchanged group (n=9). Although the chi(2) test demonstrated a statistically nonsignificant differences in RTKN expression between tumor stages Ta, T(1), and T(2) overall in the 33 human bladder carcinoma, the t test showed that there were statistically significant differences between solitary and multiple tumors, between the paired group aged younger or older than 70 years in 27 de novo bladder carcinoma patients, and between the groups with tumor larger or smaller than 2.25 cm(3). CONCLUSIONS: These results suggest that the RTKN gene is involved in bladder carcinogenesis and progression in bladder carcinoma, indicating that RTKN gene could be a molecular target in cancer therapy.


Assuntos
Povo Asiático/genética , Carcinoma de Células de Transição/genética , Carcinoma de Células de Transição/patologia , Peptídeos e Proteínas de Sinalização Intracelular/genética , Neoplasias da Bexiga Urinária/genética , Neoplasias da Bexiga Urinária/patologia , Adenocarcinoma/genética , Adenocarcinoma/patologia , Adulto , Idoso , Idoso de 80 Anos ou mais , Proteínas Reguladoras de Apoptose , Regulação para Baixo , Feminino , Proteínas de Ligação ao GTP , Regulação Neoplásica da Expressão Gênica , Humanos , Masculino , Pessoa de Meia-Idade , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Regulação para Cima
14.
Zhonghua Gan Zang Bing Za Zhi ; 12(12): 730-3, 2004 Dec.
Artigo em Chinês | MEDLINE | ID: mdl-15619339

RESUMO

OBJECTIVES: To study the effects of hepatectomized rat serum and hepatocyte growth factor (HGF) on the transdifferentiation of adult rat bone marrow stem cells (ABMSCs) into hepatic parenchymal cells. METHODS: The serum was collected from the rats 24 hours after being subjected to subtotal hepatectomy. ABMSCs were collected and cultured in DMEM/F12 (1:1) containing the hepaetectomized rat serum or HGF. The differentiated hepatocyte-like cells were labeled with CM-DiI and administrated by tail vein injection into the isogeneic rats. The cultured and injected cells were both identified by immunocytochemistry and cultured cells were assayed using RT-PCR and Western blot. RESULTS: Hepatectomized rat serum and HGF were demonstrated to have the effect of inducing transdifferentiation of ABMSCs into hepatocyte-like cells in vitro. The differentiated cells expressed albumin mRNA and albumin after 7 days++'s co-incubation. Albumin-expressing and CM-DiI positive hepatocyte-like cells were characterized in livers and spleens of the rats injected with the cultivated cells. CONCLUSION: ABMSCs could transdifferentiate into hepatic parenchymal cells by hepatectomized rat serum or HGF.


Assuntos
Células da Medula Óssea/citologia , Diferenciação Celular/fisiologia , Fator de Crescimento de Hepatócito/farmacologia , Hepatócitos/citologia , Células-Tronco/citologia , Animais , Células Cultivadas , Meios de Cultura , Hepatectomia , Regeneração Hepática , Ratos , Ratos Sprague-Dawley , Soro
15.
Scand J Immunol ; 59(6): 536-44, 2004 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-15182248

RESUMO

We investigated whether recipient dendritic cells (DCs), pretreated with nuclear factor-kappaB oligodeoxyribonucleotide decoy (NF-kappaB ODN decoy) and loaded with ultraviolet B-irradiated donor apoptotic splenocytes (Apo-SCs), were able to induce murine cardiac allograft tolerance. Heterotopic vascularized heart transplantation was performed from BALB/c to C57BL/6 mice, and recipients (C57BL/6) were given one injection of recipient DCs pretreated with NF-kappaB ODN decoy and loaded with donor (BALB/c) apoptotic SCs (decoy Apo-SCs DCs) through the portal vein at 7 days, before heart transplantation in the absence of immunosuppression. The cardiac allograft survival time and the expressive levels of intragraft cytokine genes [interleukin (IL)-2, IL-10, and interferon-gamma] were evaluated. Our results indicated that injection of decoy Apo-SCs DCs significantly prolonged vascularized heart allograft survival and led to skewing of intragraft cytokine expression towards T helper 2 (IL-10). The mechanisms can be useful for therapy of allograft rejection with minimization of systemic immunosuppression.


Assuntos
Células Dendríticas/citologia , Facilitação Imunológica de Enxerto/métodos , Sobrevivência de Enxerto/imunologia , Transplante de Coração/imunologia , Receptores do Fator de Necrose Tumoral/imunologia , Tolerância ao Transplante/imunologia , Animais , Apoptose/imunologia , Células Dendríticas/imunologia , Rejeição de Enxerto/imunologia , Interferon gama/biossíntese , Interleucina-10/biossíntese , Interleucina-2/biossíntese , Masculino , Proteínas de Membrana/imunologia , Camundongos , NF-kappa B/imunologia , Ratos , Transplante Homólogo
16.
Zhonghua Xue Ye Xue Za Zhi ; 25(5): 281-4, 2004 May.
Artigo em Chinês | MEDLINE | ID: mdl-15182536

RESUMO

OBJECTIVE: To observe the role of Panax notoginosides (PNS) in up-regulation of GATA family transcription factors, and explore intracellular signal pathway of PNS in the proliferation of hematopoietic cells. METHODS: Human bone marrow cells were incubated with different concentrations of PNS for colony-forming assay. Human cell lines HL-60, K562, CHRF-288 and Meg-01 were incubated with PNS (10 mg/L) for 14 days. The cell nuclear proteins were extracted and analyzed by Western blot with antibodies against GATA-1, GATA-2. Electrophoretic mobility shift assay (EMSA) and antibody gel supershift assay was performed using (32)P labeled GATA consensus oligonucleotide which contains binding site for GATA transcription factors. RESULTS: PNS could promote the proliferation of CFU-GM and CFU-E and induce the expression of GATA-1, GATA-2. The nuclear proteins of both GATA-1 and GATA-2 in K562, CHRF-288 and Meg-01 cells treated by PNS were increased by (1.5 - 2.8) and (2.0 - 3.1)-fold over untreated cells respectively. GATA binding activity initiated by PNS was apparently elevated to form higher density band of GATA-DNA complex. While there was no detectable change in HL-60 cells before and after PNS treatment. The predominant GATA binding complex was mainly attributable to both GATA-1 and GATA-2 proteins being in phosphorylated status. CONCLUSION: PNS can induce the synthesis of transcription factors GATA-1 and GATA-2 and enhance their DNA binding activity, which could play a role in the up-regulation of the expression genes related to proliferation and differentiation in hematopoietic cells.


Assuntos
Células da Medula Óssea/efeitos dos fármacos , Fator de Transcrição GATA1/metabolismo , Fator de Transcrição GATA2/metabolismo , Ginsenosídeos/farmacologia , Panax/química , Western Blotting , Células da Medula Óssea/metabolismo , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Relação Dose-Resposta a Droga , Células HL-60 , Humanos , Células K562 , Regulação para Cima/efeitos dos fármacos
17.
Zhongguo Shi Yan Xue Ye Xue Za Zhi ; 12(1): 16-9, 2004 Feb.
Artigo em Chinês | MEDLINE | ID: mdl-14989762

RESUMO

To observe the effects of Panax Notoginosides (PNS) on up-regulation of AP-1 family transcription factors NF-E2, c-jun and c-fos for exploring intracellular signal pathway of PNS in hematopoietic cells, four human hematopoietic cells lines including myeloid HL-60, erythroid K562, megakaryoid CHRF-288 and Meg-01 were incubated in the presence of PNS for 14 days. The nuclear protein of cells were extracted and analyzed by Western blot with antibodies against NF-E2, c-fos and c-jun. Electrophoretic mobility shift assay (EMSA) was performed by using (32)P labeled AP-1 consensus oligonucleotide which contains binding site for NF-E2, c-jun and c-fos. The results showed that the transcription factors NF-E2, c-jun and c-fos of AP-1 family could be induced by PNS. Western blot demonstrated that the nuclear protein of both NF-E2 and c-jun in four cell lines treated by PNS were increased by 1.5-2.5- and 2.0-3.0-fold over untreated cells respectively. The c-fos protein in three cell lines of K562, CHRF-288 and Meg-01 was also elevated by 2.0-3.0-fold respectively, while c-fos protein in HL-60 cells was no detectable difference after PNS treatment. EMSA results in four cell lines indicated that AP-1 binding activity initiated by PNS was apparently elevated to form higher density band of AP-1-DNA complex. In conclusion, the intracellular transcription regulation initiated by PNS was involved in transcription factors NF-E2, c-jun and c-fos of AP-1 family members, which could play an important role in the up-regulation of genes expression related to proliferation and differentiation of hematopoietic cells.


Assuntos
Proteínas de Ligação a DNA/genética , Regulação da Expressão Gênica/efeitos dos fármacos , Genes fos , Genes jun , Ginsenosídeos/farmacologia , Panax , Fator de Transcrição AP-1/metabolismo , Fatores de Transcrição/genética , DNA/metabolismo , Fatores de Ligação de DNA Eritroide Específicos , Células HL-60 , Humanos , Células K562 , Fator de Transcrição NF-E2 , Subunidade p45 do Fator de Transcrição NF-E2 , Regulação para Cima
18.
Zhonghua Fu Chan Ke Za Zhi ; 39(10): 669-74, 2004 Oct.
Artigo em Chinês | MEDLINE | ID: mdl-16144563

RESUMO

OBJECTIVE: To investigate the efficiency of antitumor immune responses induced by a controlled live dendritic cell (DC) vaccine. METHODS: DC precursors were isolated from Fischer 344 rat bone marrow and cultured with granulocyte-macrophage colony-stimulating factor and interleukin-4. The rat ovarian tumor cell line NuTu-19 was genetically modified by retroviral-mediated suicide gene (HSV(1)-TK), and the positive clones were selected using G418. Live DC vaccine was then fused with DC and NuTu-19/TK cell by polyethylene glycol. The characteristics of live DC vaccine were assayed with flow cytometry and confocal laser scanning microscopy. The specific expression of HSV(1)-TK gene in live DC vaccine was evaluated by RT-PCR and western blot. The sensitivity of live DC vaccine to ganciclovir (GCV) was evaluated by methylthiazoletetrazolium assay. In vivo, rats vaccinated twice with live DC vaccine were compared to those vaccinated with killed DC vaccine, unfused DC and NuTu-19/TK cell or phosphate buffered saline. Seven days following the last immunization, the rats were sacrificed to test the specific cytotoxic T lymphocyte (CTL) activity by lactate dehydrogenase release assay, or challenged with NuTu-19 and tumor incidence was observed. RESULTS: The fusion efficiency was approximately (23 +/- 14). Live DC vaccine displayed an up-regulated expression of major histocompatibility complex (MHC)-IIOX6 (87.6 +/- 3.4)%, costimulatory molecule B(1 - 2) (71.1 +/- 9.3)%, integrin OX 62 (68.0 +/- 7.4)%, and adhesion ICAM-1 (77.1 +/- 2.0)%, and specifically expressed HSV(1)-TK gene. Our data showed that spleen T lymphocytes from rats vaccinated with live vaccine displayed enhanced CTL activity (61.8 +/- 8.3)% contrast to that of rats vaccinated with killed vaccines (26.0 +/- 3.8)% (P < 0.05). Compared to the control groups, rats immunized with live DC vaccine demonstrated a significant delay in tumor development [(39 +/- 8) d vs (70 +/- 16) d], reduced tumor incidence (100% vs 80%) and decreased tumor volume [(806 +/- 553) mm(3) vs (89 +/- 53) mm(3), P < 0.05]. Seventy-three percent of TK-transduced live DC vaccine was killed after 7 of GCV treatment by a functional assay. CONCLUSION: The results demonstrate that DC live vaccine modified by HSV(1)-TK gene can induce efficient protective immunity and be killed by GCV.


Assuntos
Vacinas Anticâncer/imunologia , Células Dendríticas/imunologia , Herpesvirus Humano 1/enzimologia , Neoplasias Ovarianas/imunologia , Timidina Quinase/genética , Animais , Feminino , Ganciclovir/uso terapêutico , Neoplasias Ovarianas/prevenção & controle , Ratos , Ratos Endogâmicos F344 , Linfócitos T Citotóxicos/imunologia
19.
Arch Biochem Biophys ; 406(2): 173-82, 2002 Oct 15.
Artigo em Inglês | MEDLINE | ID: mdl-12361705

RESUMO

Reactive oxygen species (ROS) are important for intracellular signaling mechanisms regulating many cellular processes. Manganese superoxide dismutase (MnSOD) may regulate cell growth by changing the level of intracellular ROS. In our study, we investigated the effect of ROS on 7721 human hepatoma cell proliferation. Treatment with H2O2 (1-10 microM) or transfection with antisense MnSOD cDNA constructs significantly increased the cell proliferation. Recently, the mitogen-activated protein kinases (MAPK) and the protein kinase B (PKB) were proposed to be involved in cell growth. Accordingly, we assessed the ability of ROS to activate MAPK and PKB. PKB and extracellular signal-regulated kinase (ERK) were both rapidly and transiently activated by 10 microM H2O2, but the activities of p38 MAPK and JNK were not changed. ROS-induced PKB activation was abrogated by the phosphatidylinositol 3-kinase (PI3-K) inhibitor LY294002, suggesting that PI3-K is an upstream mediator of PKB activation in 7721 cells. Transfection with sense PKB cDNA promoted c-fos and c-jun expression in 7721 cells, suggesting that ROS may regulate c-fos and c-jun expression via the PKB pathway. Furthermore we found that exogenous H2O2 could stimulate the proliferation of PKB-AS7721 cells transfected with antisense PKB cDNA, which was partly dependent on JNK activation, suggesting that H2O2 stimulated hepatoma cell proliferation via cross-talk between the PI3-K/PKB and the JNK signaling pathways. However, insulin could stimulate 7721 cell proliferation, which is independent of cross-talk between PI3-K/PKB and JNK pathways. In addition, H2O2 did not induce the cross-talk between the PI3-K/PKB and the JNK pathways in normal liver cells. Taken together, we found that ROS regulate hepatoma cell growth via specific signaling pathways (cross-talk between PI3-K/PKB and JNK pathway) which may provide a novel clue to elucidate the mechanism of hepatoma carcinogenesis.


Assuntos
Divisão Celular/fisiologia , Sistema de Sinalização das MAP Quinases/fisiologia , Proteínas Quinases Ativadas por Mitógeno/metabolismo , Fosfatidilinositol 3-Quinases/metabolismo , Proteínas Serina-Treonina Quinases , Proteínas Proto-Oncogênicas/metabolismo , Espécies Reativas de Oxigênio/metabolismo , Receptor Cross-Talk/fisiologia , Animais , Carcinoma Hepatocelular , Humanos , Peróxido de Hidrogênio/farmacologia , Proteínas Quinases JNK Ativadas por Mitógeno , Neoplasias Hepáticas , Sistema de Sinalização das MAP Quinases/efeitos dos fármacos , Fosforilação , Proteínas Proto-Oncogênicas c-akt , Coelhos , Proteínas Recombinantes/metabolismo , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Superóxido Dismutase/genética , Superóxido Dismutase/farmacologia , Transfecção , Células Tumorais Cultivadas
20.
J Neurooncol ; 56(3): 197-208, 2002 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-12061725

RESUMO

Identification of the genes that are differentially expressed between brain tumor and normal brain tissues is important for understanding the molecular basis of these nerve system tumors and for defining possible targets for therapeutic intervention. This investigation is intended to obtain differentially expressed genes related to human glioma using cDNA microarray. Total RNA was extracted from human glioma specimens and normal brain tissues, and mRNA was obtained by oligotex chromatography. The cDNA microarray contains 4366 novel cDNA clones. The results of hybridization were scanned using computer system. Two genes selected from the results of cDNA microarray hybridization were subsequently analyzed by bio-informatic approach, Northern blot, in situ hybridization and radiation hybridization. We demonstrated that at a differentially expressed ration of two to three times, 15 cDNA clones were considered differentially expressed. Two novel full-length genes were selected for further investigation, one named human PKIbeta gene (clone 436F11, GenBank with accession number: AF225513) was over-expressed in normal brain tissues and the other named human ribosomal protein L14.22 gene (clone 507E08, GenBank with accession number: AF329277) was over-expressed in gliomas. Furthermore, the 436F11 gene was located on 6q21-q23 between the D6S304 and D6S2156 markers, while the 507E08 gene was located between the D14S1066 and D14S265 markers. We realized that cDNA microarray technology can be successfully applied to identify differentially expressed genes in human glioma. This approach is superior to routine representational difference analysis, suppression subtractive hybridization and Northern blot for detection and isolation of differentially expressed genes in different tissues. At present, we have discovered two novel full-length genes related to human glioma and their characterizations have been partially clarified.


Assuntos
Regulação Neoplásica da Expressão Gênica , Glioma/genética , Sequência de Aminoácidos , Sequência de Bases , Northern Blotting , Encéfalo/metabolismo , Mapeamento Cromossômico , Cromossomos Humanos Par 14 , Clonagem Molecular , Biologia Computacional , Primers do DNA , Humanos , Hibridização In Situ , Escore Lod , Dados de Sequência Molecular , Análise de Sequência com Séries de Oligonucleotídeos/métodos , Reação em Cadeia da Polimerase , Proteínas Quinases/química , Proteínas Quinases/genética , RNA Mensageiro/genética , Proteínas Ribossômicas/química , Proteínas Ribossômicas/genética
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