RESUMO
OBJECTIVE: To explore the expression of Foxa2 in different pathological types of gastric polyps and examine the correlation with cancerous risk. METHODS: According to computerize random number, a total of 2000 patients were selected to receive endoscopic biopsy during November 2011 to October 2012. Tissues were harvested from 170 with gastric polyps and suspicious cancerous lesions and their histological types detected. There were hyperplastic polyps(n = 35), adenomatous polyps(n = 31), fundic gland polyps(n = 42), advanced gastric cancer tissues (n = 32)and normal gastric mucosa tissues (n = 30). ABC immunohistochemical staining and reverse transcription(RT)-PCR were employed to detect the expression of Foxa2 in these different types of tissues. Imagepro plus was used for quantitative and statistical analyses. RESULTS: A low-level expression of Foxa2 was 3.6% ± 1.3% in normal gastric mucosa group. And its expreesion gradually higher in proliferative inflammatory polyp group(33.1% ± 8.0%), adenomatous polyp group (71.4% ± 1.7%) and gastric cancer group(96.3% ± 0.9%, all P < 0.05). Its expression was 35.6% ± 5.6% in fundic gland polyps, similar to that of proliferative inflammatory polyp group (P > 0.05), it was markedly lower than the gastric cancer group (P < 0.05) and higher than the normal gastric mucosa group (P < 0.05). Correlation analyses of clinicopathological parameters showed that no significant correlation existed between its expression and patient gender, age, predilection, Helicobacter. pylori infection or proton pump inhibitor used (all P > 0.05). However, the size of polyps was correlated with Foxa2 (rs = 0.69, P < 0.05). CONCLUSION: The expression level of Foxa2 in different types of gastric polyps may be used as a clinical predicator of polyps risk.
Assuntos
Pólipos Adenomatosos/metabolismo , Pólipos Adenomatosos/patologia , Fator 3-beta Nuclear de Hepatócito/metabolismo , Neoplasias Gástricas/patologia , Adulto , Idoso , Biomarcadores Tumorais/metabolismo , Estudos Transversais , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Fatores de Risco , Neoplasias Gástricas/metabolismoRESUMO
OBJECTIVE: To investigate whether oxymatrine (OM) could promote mesenchymal stem cell (MSC) therapy in CCl4-induced hepatic fibrosis (HF) in rats and to initially explore its mechanisms. METHODS: Totally 50 male SD rats were randomly divided into five groups,i.e., the normal control group, the model group, the MSC therapy group, the OM therapy group, and the MSC combined OM therapy group, 10 in each group. Except the normal control group, the HF model was duplicated by CCl4 induction. After successful modeling, rats in the MSC therapy group received 5 x10(6) MSCs by intravenous injection via caudal vein, once a week. Rats in the OM therapy group received 50 mg/kg OM by intramuscular injection, three times a week. Rats in MSC combined OM therapy group received 5 x 10(6) MSCs by intravenous injection via caudal vein, once a week and 50 mg/kg OM by intramuscular injection three times a week. Equal volume of normal saline was given to those in the normal control group and the model group. All medication lasted for 8 weeks. Serum levels of ALT and AST were detected 8 weeks later. The hepatic histopathological injury and extracellular matrix deposit were assessed using HE and Masson staining. Expressions of serum interleukin-4 (IL-4) and interleukin-10 (IL-10) were detected using enzyme linked immunosorbent assay (ELISA). RESULTS: (1) Compared with the normal control group, serum levels of ALT and AST significantly increased in the model group (P < 0.05). Compared with the model group, serum levels of ALT and AST significantly decreased in the OM therapy group, the MSC therapy group, and the MSC combined OM therapy group at the end of 8 weeks of treatment (P < 0.05). But serum levels of ALT and AST were significantly lower in the MSC combined OM therapy group than in the OM therapy group and the MSC therapy group (P < 0.05). (2) Compared with the model group, the hepatic injury was significantly lessened and the area of extracellular matrix deposit was significantly reduced in the OM therapy group, the MSC therapy group, and the MSC combined OM therapy group (P < 0.05). Besides, they wer more significant in the MSC combined OM therapy group (P < 0.05). (3) Compared with the model group, the serum IL-4 level was significantly higher in the MSC therapy group and the MSC combined MO group (P < 0.05). It was higher in the MSC combined MO group (P < 0.05). Although the serum IL-4 level also increased in the OM therapy group, but with no statistical difference (P > 0.05). (4) The serum IL-10 level significantly increased in the OM therapy group, the MSC therapy group, and the MSC combined OM therapy group (P < 0.05), and it was the highest in the MSC combined OM therapy group among the three groups (P < 0.05). (5) Two-photon fluorescence imaging showed no signals of MSCs in liver with or without OM injection. CONCLUSION: OM could promote mesenchymal stem cell therapy in hepatic fibrosis rats, which might be involved in increasing serum levels of IL-4 and IL-10.
Assuntos
Alcaloides/uso terapêutico , Cirrose Hepática Experimental/terapia , Transplante de Células-Tronco Mesenquimais , Quinolizinas/uso terapêutico , Animais , Interleucina-10/sangue , Interleucina-4/sangue , Masculino , Ratos , Ratos Sprague-DawleyRESUMO
OBJECTIVE: To analyze differentially expressed proteins of hepatic stellate cells (HSCs) treated with oxymatrine (OMT) liposomes, thus further exploring the molecular mechanism of OMT liposomes for treating liver fibrosis. METHODS: A rat model of CCl4 induced chronic liver fibrosis was established. HSCs were perfusion isolated from modeled SD rats and cultured in vitro . Passage 2 HSCs were divided into the model group (Group A), the OMT-liposome-treated group (Group B), and the liposome-treated control group (Group C). HSCs from normal rats were taken as the normal control group (Group D). The total proteins of HSCs cells were extracted from Group B and D after 7 days of treatment, and separated with isoelectrofocusing two-dimensional electrophoresis (2-DE). A 2-DE system was established to analyze the differences in the protein profile between Group B and Group C. Tow protein dots with most obvious difference were selected to determine the structures and functions of different proteins using peptide mass fingerprinting (PMF). RESULTS: (1) The total number bf proteins decreased after treated with OMT liposomes, with 864 spots before treatment and 756 spots after treatment, and the matching rate was 63%. (2) According to 2-DE results, 10 differential protein spots were found by image analysis of magnifying images in local regions. (3) Two most differently expressed proteins were identified to be ATM (46. 236 kD) and Miz1 (54. 051 kD) by PMF and SWISS-PROT protein database retrieval. CONCLUSION: Action of OMT liposomes on HSCs of rats with chronic liver fibrosis caused different protein expressions, which might be involved in the signaling pathways of inducing the apoptosis of HSCs.
Assuntos
Alcaloides/farmacologia , Células Estreladas do Fígado/efeitos dos fármacos , Proteoma/metabolismo , Quinolizinas/farmacologia , Animais , Eletroforese em Gel Bidimensional , Células Estreladas do Fígado/citologia , Células Estreladas do Fígado/metabolismo , Lipossomos , Cirrose Hepática Experimental/metabolismo , Masculino , Ratos , Ratos Sprague-DawleyRESUMO
Tumor immune tolerance plays a critical role in tumor cell survival; the establishment of tumor immune tolerance is incompletely understood yet. Integrin alphavbeta6 (avb6) is involved in tumor growth and metastasis. This study aimed to observe the effect of avb6 on the development of tumor tolerance in colorectal cancer (CRC). In this study, 28 CRC patients were recruited. The frequencies of tolerogenic dendritic cells (TolDC), regulatory T cells (Treg), and CD8+ T cells in surgically removed CRC tissue were assessed by flow cytometry. The levels of avb6 in CRC tissue were measured by enzyme-linked immunoassay (ELISA). The effect of avb6 on inducing TolDCs and Tregs was evaluated with the cell culture model. The results showed that in surgically removed CRC tissue, we detected higher frequencies of TolDC and Tregs, lower frequency CD8+ T cells and high levels of avb6 as compared with non-CRC tissue. CRC protein extracts could induce TolDC development that could be blocked by anti-avb6 antibody. CRC-derived DCs could convert naïve CD4+ T cells to Tregs. Peripheral CD8+ T cells from CRC patients still retained the ability to produce granzyme B and to proliferate in response to CRC tumor antigen in culture that was abolished by the presence of CRC-derived Tregs. We conclude that CRC-derived avb6 is involved in the establishment of tumor immune tolerance in local tissues.
Assuntos
Antígenos de Neoplasias/imunologia , Neoplasias Colorretais/imunologia , Tolerância Imunológica/imunologia , Integrinas/imunologia , Adulto , Idoso , Idoso de 80 Anos ou mais , Antígenos de Neoplasias/metabolismo , Western Blotting , Linfócitos T CD4-Positivos/imunologia , Linfócitos T CD4-Positivos/metabolismo , Linfócitos T CD8-Positivos/imunologia , Linfócitos T CD8-Positivos/metabolismo , Proliferação de Células , Células Cultivadas , Neoplasias Colorretais/metabolismo , Neoplasias Colorretais/patologia , Células Dendríticas/imunologia , Células Dendríticas/metabolismo , Ensaio de Imunoadsorção Enzimática , Feminino , Citometria de Fluxo , Granzimas/imunologia , Granzimas/metabolismo , Humanos , Integrinas/metabolismo , Masculino , Pessoa de Meia-Idade , Linfócitos T Reguladores/imunologia , Linfócitos T Reguladores/metabolismoRESUMO
BACKGROUND AND AIM: As a newly identified subset of T helper cells, T-helper 17 cells (Th17) are major mediators of inflammation-associated disease. Some reports have revealed significantly increased Th17 cells in hepatitis B virus-infected patients, and a recent study has demonstrated that hepatitis C virus (HCV)-specific Th17 cells can be induced in vitro and regulated by transforming growth factor-ß. This study attempted to characterize the role of Th17 cells in the disease progression of chronic hepatitis C (CHC). METHODS: The current study enrolled 53 patients with CHC and 23 healthy controls, in which the circulating and liver-infiltrating Th17 cells were monitored. RESULTS: We found that CHC patients had increased proportions of both circulating and liver-infiltrating Th17 cells compared to healthy individuals, and both measures of Th17 cells were correlated with severity of liver inflammation. We further demonstrated that the HCV-specific Th17 cells were correlated with liver damage but not HCV viral replication. CONCLUSIONS: Such a correlation between the severity of liver damage of CHC and Th17 cells illustrated in this study sheds some light on the understanding of the pathogenesis of CHC.
Assuntos
Hepacivirus/imunologia , Hepatite C Crônica/imunologia , Fígado/imunologia , Células Th17/imunologia , Alanina Transaminase/sangue , Biomarcadores/sangue , Biópsia , Contagem de Linfócito CD4 , Estudos de Casos e Controles , Células Cultivadas , China , Citometria de Fluxo , Hepacivirus/genética , Hepacivirus/patogenicidade , Hepatite C Crônica/diagnóstico , Hepatite C Crônica/patologia , Humanos , Interleucina-17/sangue , Fígado/patologia , Fígado/virologia , RNA Viral/sangue , Índice de Gravidade de Doença , Células Th17/patologia , Células Th17/virologia , Carga Viral , Replicação ViralRESUMO
OBJECTIVE: To assess the differential diagnostic value of serum intestinal fatty acid binding protein (I-FABP) in distinguishing intestinal ischemia patients from acute abdomen patients. METHODS: A total of 151 patients with acute abdomen and 17 healthy controls from the PLA General Hospital were enrolled from November, 2009 to August, 2011. Serum I-FABP levels were measured by ELISA. According to the ROC curve, the cut-off value, sensitivity, specificity, positive likelihood ratio (PLR), negative likelihood ratio (NLR), positive predictive value (PPV) and negative predictive value (NPV) were calculated. RESULTS: Of the 151 acute abdomen patients, there were 24 intestinal ischemia patients and 127 without intestinal ischemia. Serum I-FABP level in intestinal ischemia group [(109.67 ± 48.82) µg/L] was significantly higher than those in patients without intestinal ischemia [(36.78 ± 11.25) µg/L] and healthy controls[(8.33 ± 6.25) µg/L](all P values < 0.01). The serum I-FABP cut-off value for the diagnosis of intestinal ischemia was 87.52 µg/L. Serum I-FABP was efficient in terms of sensitivity (0.762), NPV(0.963), PLR(3.05) and NLR (0.24) in the diagnosis of intestinal ischemia. CONCLUSION: I-FABP is potentially useful for discriminating intestinal ischemia from acute abdomen.
Assuntos
Abdome Agudo/diagnóstico , Proteínas de Ligação a Ácido Graxo/sangue , Intestinos/fisiopatologia , Isquemia/diagnóstico , Abdome Agudo/sangue , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Estudos de Casos e Controles , Criança , Diagnóstico Diferencial , Feminino , Humanos , Intestinos/irrigação sanguínea , Isquemia/sangue , Masculino , Pessoa de Meia-Idade , Valor Preditivo dos Testes , Estudos Prospectivos , Curva ROC , Sensibilidade e Especificidade , Adulto JovemRESUMO
OBJECTIVE: To investigate the related factors of recurrence of early gastric cancer (EGC) after endoscopic resection. METHODS: Clinicopathologic data of 169 patients with EGC who underwent endoscopic resection and periodically followed up by the Chinese PLA General hospital were analyzed retrospectively. RESULTS: During a follow-up of 13 - 57 months (median time 24.5 months), 12 patients had gastric cancer again and the recurrence rate was 7.1% (12/169). The recurrence time varied from 3 to 36 (28 ± 23) months and the median time was 18 months. The recurrence rates of 0.5 year, 1(st) year, 2(nd) year and 3(rd) year were 1.18% (2/169), 3.55% (6/169), 9.91% (11/111) and 12.24% (12/98), respectively. Eleven patients had gastric cancer again within 2 years after resection. Undifferentiated histology (including poorly differentiated carcinoma and signet ring cell carcinoma), submucosal infiltration and lymphovascular invasion of the primary lesion of EGC were related to the postsurgical recurrence (all P < 0.05). CONCLUSION: Most recurrence of EGC occurred within 2 years after endoscopic resection and is related with undifferentiated histology, submucosal infiltration and lymphovascular invasion. It is important for these patients to receive endoscopy follow up.
Assuntos
Adenocarcinoma/patologia , Recidiva Local de Neoplasia/patologia , Neoplasias Gástricas/patologia , Adenocarcinoma/cirurgia , Adulto , Idoso , Idoso de 80 Anos ou mais , Endoscopia , Feminino , Gastrectomia/métodos , Humanos , Masculino , Pessoa de Meia-Idade , Estudos Retrospectivos , Neoplasias Gástricas/cirurgiaRESUMO
OBJECTIVE: To analyze clinical characteristics of patients with gastrointestinal bleeding (GIB) and the death-related risk factors. METHODS: A retrospective analysis was conducted in 414 patients hospitalized for GIB during a 16-year period of 1994 to 2009. Logistic regression analysis identified predictors of mortality. RESULTS: The mean age of the 414 patients is 83.5 years old, ranging from 65 to 96 years old. The main causes of GIB were peptic ulcer (33.1%, 137/414), gastroduodenal mucosal erosion (28.5%, 118/414) and tumor (21.0%, 87/414). The main symptom was melena (71.0%, 294/414). Drugs that induced GIB were mainly non-steroidal anti-inflammatory drugs, including aspirin (11.1%, 46/414), acetaminophen (8.9%, 37/414) and indomethacin (1.9%, 8/414). 14% of patients (58/414) died of GIB in 30 days. The proportion of drug-induced GIB and gastroduodenal mucosal erosion caused GIB had increased significantly during the period of 2004 to 2009 (P < 0.05). Analysis of 30-day mortality risk showed advanced age, low diastolic blood pressure, high heart rate, low hemoglobin levels at presentation and hemorrhage volume in dead GIB elderly patients were significantly different compared with GIB elderly patients alive. Presence of severe comorbidity (heart failure and renal failure) and caused by cirrhosis and portal hypertension in GIB elderly patients were the only independent predictors of 30-day mortality (P < 0.001). CONCLUSIONS: Death of GIB patients occurred predominantly in elderly patients with severe comorbidities and systemic conditions at presentation.
Assuntos
Hemorragia Gastrointestinal/epidemiologia , Hemorragia Gastrointestinal/mortalidade , Fatores Etários , Idoso , Idoso de 80 Anos ou mais , Feminino , Hemorragia Gastrointestinal/etiologia , Humanos , Modelos Logísticos , Masculino , Estudos Retrospectivos , Fatores de RiscoRESUMO
OBJECTIVE: Aberrant expression of immunoglobulin (Ig) by cancer cells has been documented in a number of malignant tumors but its biological significance is unclear. Cancer cells overexpress anti-apoptotic molecules such as Bcl-xL. The present study aimed to examine the role of expression of Ig light-chain Igk and Iglambda in maintaining the high levels of Bcl-xL in colorectal cancer cells. MATERIAL AND METHODS: Thirty patients with colorectal cancer were recruited to this study. Expression of Igk, Iglambda and Bcl-xL in surgically removed cancer tissue was examined by immunohistochemistry and/or flow cytometry. Using the HT29 cell line as a study platform, RNA interference (RNAi) was employed to knock out the genes of Igk and Iglambda in the cancer cell line; the expression of Bcl-xL in HT29 cells was subsequently analyzed. RESULTS: Human colorectal cancer cells, but not normal colorectal tissue, expressed both Igk and Iglambda in the cytoplasm. High levels of Bcl-xL were detected in cancer cells. Using RNAi to knock out the genes of Igk and/or Iglambda, Bcl-xL expression in HT29 cells was significantly suppressed and the cells became apoptotic. CONCLUSION: The results suggest that expression of Igk and Iglambda is required to stabilize Bcl-xL expression in cancer cells.
Assuntos
Neoplasias Colorretais/metabolismo , Cadeias lambda de Imunoglobulina/metabolismo , Imunoglobulinas/metabolismo , Fatores Imunológicos/metabolismo , Proteína bcl-X/metabolismo , Adulto , Idoso , Biomarcadores Tumorais/metabolismo , Western Blotting , Neoplasias Colorretais/genética , Neoplasias Colorretais/patologia , Feminino , Citometria de Fluxo , Humanos , Cadeias lambda de Imunoglobulina/genética , Imunoglobulinas/genética , Imuno-Histoquímica , Fatores Imunológicos/genética , Masculino , Pessoa de Meia-Idade , Invasividade Neoplásica , Estadiamento de Neoplasias , Proteína bcl-X/genéticaRESUMO
AIM: To analyze the expression profiles of a human gastric-cancer-related gene, GCRG123, in human gastric signet-ring cell carcinoma tissues, and to perform bioinformatics analysis on GCRG123. METHODS: In situ hybridization was used to explore the GCRG123 expression pattern in paraffin-embedded gastric tissues, including 15 cases of signet-ring cell carcinoma, 15 of intestinal-type adenocarcinoma, and 15 of normal gastric mucosa. Northern blotting was used to analyze the differences in GCRG123 expression between stomach signet-ring cell carcinoma and intestinal-type adenocarcinoma tissues. Online software, including BLAST, Multalin and BLAT, were applied for bioinformatics analysis. National Center for Biotechnology Information (NCBI) and the University of California Santa Cruz (UCSC) databases were used for the analyses. RESULTS: The in situ hybridization signal appeared as blue precipitates restricted to the cytoplasm. Ten out of 15 cases of gastric signet ring cell carcinoma, normal gastric mucosal epithelium and pyloric glands showed high GCRG123 expression. Low GCRG123 expression was observed in gastric intestinal-type adenocarcinoma and normal gastric glands. Northern blotting revealed that GCRG123 was up-regulated in signet-ring cell carcinoma tissue but down-regulated in intestinal-type adenocarcinoma tissue. BLAST and Multalin analyses revealed that the GCRG123 sequence had 92% similarity with the ORF2 sequence of human long interspersed nuclear element retrotransposons (LINE-1, L1). BLAT analysis indicated that GCRG123 mapped to all chromosomes. GCRG123 was found to integrate in the intron-17 and -23 of Rb, 5' flanking region of IL-2 and clotting factor IX genes. CONCLUSION: GCRG123, an active member of the L1 family, was up-regulated in human gastric signet-ring cell carcinoma.
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Carcinoma de Células em Anel de Sinete/genética , Regulação Neoplásica da Expressão Gênica , Laminas/genética , Neoplasias Gástricas/genética , Humanos , Elementos Nucleotídeos Longos e Dispersos , Dados de Sequência Molecular , Regulação para Cima/genéticaRESUMO
OBJECTIVE: Some members of the S100 gene family have been suggested to be associated with cancer development and metastasis. Our previous cDNA micro-array studies have showed S100A6 expression is elevated in gastric cancer compared with that in paired normal mucosa. To validate our previous results and further investigate the possible role of S100A6 gene in gastric cancer, we carried out this detailed S100A6 expression analysis in more matched gastric cancer samples. METHODS: S100A6 expression was detected in 20 paired fresh surgical samples of gastric tumor tissue and matched non-cancerous mucosa by QRT-PCR. A gastric cancer tissue microarray (TMA) containing 1020 duplicate matched normal mucosa, gastric cancer tissue and metastatic lymph node tissue cores from 208 gastric cancer patients was constructed. S100A6 expression was detected by immunohistochemistry and the correlation between S100A6 expression with clinicopathological factors and survival was analyzed. RESULTS: As quantitated by QRT-PCR, S100A6 transcript level was elevated in 73.7% of the primary cancer lesions with an average 2.25-fold up-regulation than that in matched non-neoplastic mucosa. As displayed by immunohistochemistry, the positive rate of S100A6 in non-neoplastic mucosa, tumor lesions and metastatic lymph nodes was 34.3%, 84.1% and 90.9%, respectively. S100A6 expression level in cancer and metastatic lymph node was significantly higher than their matched non-neoplastic mucosa (P < 0.05). 65.5% of patients showed an increased S100A6 expression in cancer tissue compared with that in matched normal mucosa. S100A6 overexpression was associated with larger tumor size and deeper invasion (P = 0.022 and P = 0.009). No evidence was found for an association between S100A6 expression level and other variables, including tumor grade, nodal metastases, and TNM stage. There was no association between S100A6 expression level and survival. But compared with paired non-neoplastic mucosa, an increased S100A6 expression in tumor lesion predicated a decreasing suvival if compared with a decreased S100A6 expression, though the difference was statistically not significant. CONCLUSION: Elevated expression of S100A6 gene may be an early event in the development and progression of gastric cancer. Further study of this gene may be helpful for understanding the nature of gastric carcinoma.
Assuntos
Proteínas de Ciclo Celular/metabolismo , Proteínas S100/metabolismo , Neoplasias Gástricas/metabolismo , Seguimentos , Mucosa Gástrica/metabolismo , Regulação Neoplásica da Expressão Gênica , Humanos , Metástase Linfática , Invasividade Neoplásica , Estadiamento de Neoplasias , RNA Mensageiro/metabolismo , Proteína A6 Ligante de Cálcio S100 , Neoplasias Gástricas/patologia , Neoplasias Gástricas/cirurgia , Taxa de Sobrevida , Carga Tumoral , Regulação para CimaRESUMO
OBJECTIVE: To investigate the effect of gene GCRG213 siRNA transfection into gastric cancer cell line MKN45 cells. METHODS: Two pairs of DNA sequences containing small hairpin structure to GCRG213 were designed and synthesized. The complement form was obtained by annealing and inserted into RNAi expression vector IMG-800. They are IMG-800-1 and IMG-800-2 correspondingly. The recombinant plasmid IMG-800-1, IMG-800-2 and the vector IMG-800 were separately transfected into MKN45 cells conducted by lipofectamine 2000. After G418 selecting, the cells were transfected steadily. Expression of GCRG213 was detected by semi-quantitative RT-PCR and Western Blot. The growth graph of six steady transfected cell cultures was protracted by cell counting. FACS was used to detect the cell cycle, and Annexin V FITC/PI double labeling were used to detect the effects on cell apoptosis in the above-mentioned cells. The clone formation rate in plate and in nude mice was tested to investigate the tumorigenic characteristics of the six steadily transfected cells in vitro and vivo. RESULTS: Through sequencing, two pairs of DNA sequences containing small hairpin structure to GCRG213 were proved to be successfully cloned into siRNA expression vector IMG-800, correspondingly called IMG-800-1 and IMG-800-2. The recombinant plasmid IMG-800-1, IMG-800-2 and vector IMG-800 were transfected separately into MKN45 cells conducted by lipofectamine 2000. After G418 selecting, the cells were transfected steadily. Transfecting the siRNA vector (IMG-800-1, IMG-800-2 ) into the MKN45 cells significantly decreased the expression of GCRG213, at both mRNA and protein levels. The growth graph showed that the growth of IMG-800-1 and IMG-800-2 transfected cells were slower than that of vector transfected cells. The proportion of cells in G2/M and/or S phase decreased in the cells transfected with IMG-800-1 and IMG-800-2 and cell apoptosis increased. The average clone formation rate in vitro decreased in the cells transfected with IMG-800-1 and IMG-800-2, compared with those transfected with vector. In vivo, the time of tumor formation of IMG-800-1 and IMG - 800-2 transducted cells in nude mice was prolonged and the tumor size was smaller. CONCLUSION: GCRG213 SiRNA transfection may induce inhibition of growth and proliferation of tumor cells, promote cell apoptosis, and inhibit the tumorigenicity in vitro and vivo.
Assuntos
Adenocarcinoma/patologia , Hormônios Peptídicos/biossíntese , RNA Interferente Pequeno/genética , Neoplasias Gástricas/patologia , Adenocarcinoma/genética , Adenocarcinoma/metabolismo , Animais , Apoptose , Western Blotting , Ciclo Celular , Linhagem Celular Tumoral , Proliferação de Células , Feminino , Vetores Genéticos , Humanos , Camundongos , Camundongos Nus , Transplante de Neoplasias , Hormônios Peptídicos/genética , Interferência de RNA , RNA Mensageiro/biossíntese , RNA Mensageiro/genética , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Neoplasias Gástricas/genética , Neoplasias Gástricas/metabolismo , Transfecção , Transplante HeterólogoRESUMO
Radiation-induced lung injury (RILI) is a major clinical complication for radiotherapy in thoracic tumors. An immediate effect of lung irradiation is the generation of reactive oxygen that can produce oxidative damage to DNA, lipids, and proteins resulting in lung cell injury or death. Currently, the medical management of RILI remains supportive. Therefore, there is an urgent need for the development of countermeasures. The present study aimed to evaluate the protective effect of manganese superoxide dismutase (MnSOD) gene-modified mesenchymal stem cells (MSCs) to facilitate the improved recovery of RILI. Here, nonobese diabetic/severe combined immunodeficiency mice received a 13 Gy dose of whole-thorax irradiation, and were then transfused intravenously with MnSOD-MSCs and monitored for 30 days. Lung histopathologic analysis, plasma levels of inflammatory cytokines (interleukin [IL]-1, IL-6, IL-10, and tumor necrosis factor-α), profibrotic factor transforming growth factor-ß1, and the oxidative stress factor (hydroxyproline) were evaluated after MnSOD-MSC transplant. Apoptotic rates were evaluated by terminal deoxynucleotidyl transferase-mediated nick-end labeling immunohistochemical method. Colonization and differentiation of MnSOD-MSCs in the irradiated lung were analyzed by immunofluorescence staining. Consequently, systemic administration of MnSOD-MSCs significantly attenuated lung inflammation, ameliorated lung damage, and protected the lung cells from apoptosis. MnSOD-MSCs could differentiate into epithelial-like cells in vivo. MnSOD-MSCs were effective in modulating RILI in mice and had great potential for accelerating from bench to bedside.
Assuntos
Lentivirus/genética , Lesão Pulmonar/terapia , Transplante de Células-Tronco Mesenquimais , Células-Tronco Mesenquimais/metabolismo , Superóxido Dismutase/genética , Administração Intravenosa , Animais , Apoptose/genética , Líquido da Lavagem Broncoalveolar , Raios gama/efeitos adversos , Expressão Gênica , Genes Reporter , Vetores Genéticos/química , Vetores Genéticos/metabolismo , Proteínas de Fluorescência Verde/genética , Proteínas de Fluorescência Verde/metabolismo , Humanos , Interleucina-1/genética , Interleucina-1/metabolismo , Interleucina-10/genética , Interleucina-10/metabolismo , Interleucina-6/genética , Interleucina-6/metabolismo , Lentivirus/metabolismo , Pulmão/metabolismo , Pulmão/patologia , Pulmão/efeitos da radiação , Lesão Pulmonar/etiologia , Lesão Pulmonar/metabolismo , Lesão Pulmonar/patologia , Masculino , Células-Tronco Mesenquimais/citologia , Células-Tronco Mesenquimais/imunologia , Camundongos , Camundongos SCID , Superóxido Dismutase/metabolismo , Transgenes , Transplante Heterólogo , Fator de Necrose Tumoral alfa/genética , Fator de Necrose Tumoral alfa/metabolismoRESUMO
AIM: To clone genes that may predispose us to human gastric cancer and to analyze it's expression in gastric tissues. METHODS: Specimens of paired tumor, paratumor and normal gastric mucosa tissues collected from fifteen patients who suffered from stomach antrum adenocarcinoma were used for analysis. Seven out of the fifteen cases were first studied by fluorescent differential display reverse transcription polymerase chain reaction (DDTR-PCR) analysis. The differentially expressed bands of interest were cloned, analyzed by Northern blot, sequencing and RT-PCR. Through BLAST, the sequencing results were compared with GenBank database for homology analysis. In situ hybridization with DIG-labeled cRNA probes was used to analyze the expression of interesting cDNA bands in paraffin embedded paired normal gastric mucosa and cancer tissues isolated from 30 gastric adenocarcinoma patients. RESULTS: DDRT-PCR showed that one of the interesting cDNA bands, which was named W2, expressed much higher in all seven tested tumor and paratumor samples than in their normal counterparts, it was sub-cloned into a pGEM-T Easy vector. Two subclones were subsequently obtained. One of the subclone, GCRG224, was studied further. The sequencing result showed that GCRG224 consisted of 1 159 base pairs and had one open reading frame (ORF). It located at human chromosome 11q14. No homologue was found in GenBank database with GCRG224-ORF. This nucleotide sequence data were submitted to GenBank with accession No. AF438406. RT-PCR showed that GCRG224 expressed higher in 11/15 gastric cancer tissues than in non-tumor tissues. However, the result of Northern blot analysis showed a higher GCRG224 expression in the non-tumor tissue than in the tumor one. Human multiple tissue Northern blot analysis revealed that GCRG224 also expressed in human normal colon tissue, and peripheral blood leukocyte. In situ hybridization analysis showed that only 5/30 adenocarcinoma, 3/18 dysplasia and 6/18 intestinal metaplasia showed higher GCRG224 expression level than the normal gastric glands. However, GCRG224 was over-expressed predominantly in 26/30 cases of normal mucosal epithelium. CONCLUSION: A novel gene named GCRG224 was identified from human gastric mucosal tissue. It overexpressed in almost all gastric mucosal epithelium but only a small portion of cancer and precancerous leisions. The role of GCRG224 expression in gastric epithelium needs further study.
Assuntos
Adenocarcinoma/genética , Mucosa Gástrica/metabolismo , Regulação Neoplásica da Expressão Gênica , Proteínas/metabolismo , Neoplasias Gástricas/genética , Adenocarcinoma/metabolismo , Adenocarcinoma/patologia , Adulto , Idoso , Feminino , Perfilação da Expressão Gênica , Humanos , Hibridização In Situ , Masculino , Pessoa de Meia-Idade , Dados de Sequência Molecular , Proteínas de Neoplasias/genética , Proteínas de Neoplasias/metabolismo , Reação em Cadeia da Polimerase/métodos , Lesões Pré-Cancerosas , Proteínas/genética , RNA Mensageiro/metabolismo , Neoplasias Gástricas/metabolismo , Neoplasias Gástricas/patologiaRESUMO
AIM: To identify the gene that may predispose to human gastric cancer and to analyze its expression in gastric cancer and non-tumorous gastric mucosa. METHODS: Cancer, para-tumor, and non-tumor gastric tissues were studied for gene expression profile using fluorescent differential display reverse transcription polymerase chain reaction (DDRT-PCR). The differentially expressed bands of interest were analyzed by cloning, Northern blotting, and sequencing. The sequencing results were compared with the GenBank database for homology and conserved domain analysis. In situ hybridization with DIG-labeled cRNA probes was used to detect the expression of gene in paraffin embedded gastric adenocarcinoma and non-cancerous tissues. RESULTS: A gene expressed higher in tumor and para-tumor tissues than in their non-tumor counterparts of all 7 tested gastric adenocarcinoma patients was identified by means of DDRT-PCR analysis. It was named GCRG213 (gastric cancer related gene 213). Northern blot confirmed the differential expression. GCRG213 (GenBank No. AY053451) consisted of 1094 base pairs with an open reading frame (ORF) which encoded 142 amino acids. The deduced amino acid sequence contained a putative conserved domain, apurinic/apyrimidinic endonuclease (APE). In situ hybridization analysis showed that GCRG213 was expressed higher in gastric cancer tissues than in their corresponding non-tumor ones. Precancerous leisions of gastric adenocarcinoma showed a high GCRG213 expression, too. No difference of the expression patterns was found between the early and advanced gastric cancer. CONCLUSION: A gene named GCRG213 was identified in human gastric adenocarcinoma. It encoded an APE-like protein which was probably a new member of the APE family. GCRG213 was over-expressed not only in gastric cancer, but also in its precancerous leisions. The role of GCRG213 expression in carcinogenesis needs further study.
Assuntos
Endonucleases/genética , Neoplasias Gástricas/genética , Regulação para Cima , Adulto , Idoso , Sequência de Aminoácidos/genética , Sequência de Bases/genética , DNA Complementar/genética , Feminino , Humanos , Hibridização In Situ , Masculino , Pessoa de Meia-Idade , Dados de Sequência Molecular , Hormônios Peptídicos , RNA Mensageiro/metabolismo , Neoplasias Gástricas/metabolismoRESUMO
AIM: To improve the prevention and treatment of senile patients with colorectal cancer by evaluating the importance of colonoscopy in clinical screening and follow-up. METHODS: Clinical screening of colonoscopy was performed for 2196 patients aged 60-90 years old according to the protocol,and 1740 of them (79.2%) were followed-up. RESULTS: Colorectal cancer was found in 52 patients, and the detectable rate was 2.4%. Among them, 19 were diagnosed as early colorectal cancer, accounting for 36.5% of the detected colorectal cancer. Among the followed-up patients, early colorectal cancer was found in 9, accounting for 45.0% of the detected colorectal cancer. The resectable rate and 5 years survival rate of colorectal cancer were 97.7% and 80.9% respectively. The incidence of complication was 0.05%, and the successful rate of cecum intubation was 98.9%. CONCLUSION: Colonoscopic screening and follow-up of the elderly for colorectal cancer and pre-cancerous lesion (adenomatoid polyp) can increase the detectable rate of early colorectal cancer and improve its prevention and treatment.
Assuntos
Colonoscopia , Neoplasias Colorretais/diagnóstico , Programas de Rastreamento , Idoso , Idoso de 80 Anos ou mais , Neoplasias Colorretais/epidemiologia , Neoplasias Colorretais/mortalidade , Neoplasias Colorretais/cirurgia , Humanos , Pessoa de Meia-Idade , Taxa de SobrevidaRESUMO
OBJECTIVE: To study the relation between dendritic cell (DC) infiltration and clinicopathologic parameters, biologic characteristics and prognosis of progressing gastric cancer. METHODS: The development of apoptotic cell death (apoptotic index, AI) in 61 progressing gastric carcinoma tissues was analyzed by terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate biotin nick end labeling (TUNEL) method. The PCNA labeling index (PCNA-LI), density of dendritic cells in the tumor were detected by immunohistochemical method by the LSAB kit using antibody against S-100 protein and PC-10. RESULTS: DC infiltration was negatively correlated with lymph node metastasis, clinical stage and PCNA-LI, but positively with AI. The DCs in gastric cancer groups with and without lymph node metastasis were (5.63 +/- 4.37)/HPF and (8.51 +/- 5.57)/HPF with difference significant (P < 0.05). The DC infiltration in I, II, III stage lesions were (11.23 +/- 6.05)/HPF, (6.28 +/- 4.37)/HPF and (5.53 +/- 5.19)/HPF also with differences significant (P < 0.01). The PCNA-LI was significantly higher in the low DC group (57.10% +/- 14.18%) than that of high DC group (48.15% +/- 10.59%, P < 0.01). AI findings were 3.77% +/- 1.26% and 2.95% +/- 1.07% in the high and low DC groups (P < 0.01). A positive correlation was observed between DC infiltration and AI (r = 0.39, P < 0.01) whereas a negative correlation between DC infiltration and PCNA-LI (r = -0.47, P < 0.01). The prognosis of high DC infiltration patients was significantly better than those with low ones. CONCLUSION: The infiltrating dendritic cells in and around tumor, representing the local immune status of the host, may play an important role in immunological defense mechanism of host versus tumor. Dendritic cells may inhibit the proliferation and induce the apoptosis of the tumor cells, thus affecting the clinical features and improve the prognosis of gastric carcinoma.
Assuntos
Células Dendríticas/fisiologia , Neoplasias Gástricas/patologia , Adulto , Idoso , Idoso de 80 Anos ou mais , Apoptose , Divisão Celular , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Prognóstico , Antígeno Nuclear de Célula em Proliferação/análise , Neoplasias Gástricas/mortalidade , Taxa de SobrevidaRESUMO
OBJECTIVE: To study the clinical features and proper treatment of 38 elderly patients with early double primary cancers. METHODS: Thirty-eight elderly patients with early double primary cancers treated from January 1980 to March 2003 were retrospectively reviewed for involved organs, treatment and prognosis. RESULTS: Digestive tract was the most frequently involved, followed by urogenital system and lung. Long-term results of endoscopic mucosal resection (EMR), operation and radiotherapy were superior to other methods. The prognosis of gastrointestinal carcinoma was better than that of prostate carcinoma and hematopoietic system. The operation rate decreased with increasing age. The 5-year survival rates of EMR, operation and radiotherapy were 85.7%, 71.1% and 75.0%, respectively. The medium survival time was 120 months in first cancer and 39 months in the second primary cancer. The 5-year survival rates of the first cancer and second primary cancer were 88.6% and 53.8%. CONCLUSION: Yearly follow-up for elderly patients with endoscopy, beta ultrasonic scan and X-ray contribute to finding of early double primary cancers. Operation is the best treatment of early double primary cancers. Endoscopic mucosal resection is especially suitable for old patients with digestive tract and bladder cancer.
Assuntos
Neoplasias Colorretais , Neoplasias Pulmonares , Neoplasias Primárias Múltiplas , Neoplasias da Próstata , Neoplasias Gástricas , Idoso , Idoso de 80 Anos ou mais , Neoplasias Colorretais/diagnóstico , Neoplasias Colorretais/radioterapia , Neoplasias Colorretais/cirurgia , Endoscopia do Sistema Digestório , Feminino , Seguimentos , Humanos , Neoplasias Pulmonares/radioterapia , Neoplasias Pulmonares/cirurgia , Masculino , Pessoa de Meia-Idade , Neoplasias da Próstata/radioterapia , Neoplasias da Próstata/cirurgia , Estudos Retrospectivos , Neoplasias Gástricas/radioterapia , Neoplasias Gástricas/cirurgia , Taxa de SobrevidaRESUMO
OBJECTIVE: To investigate the gene expression profile of human gastric adenocarcinoma by means of cDNA microarray and to analyze its biological significance. METHODS: Paired tumor and non-tumor specimens from 18 cases of advanced gastric adenocarcinoma were studied. Total RNA was isolated and labeled by reverse transcription reaction with cy5 and cy3 for cDNA probe. cDNA microarrays containing 148 genes were hybridized with labeled cDNA probe. Data from cDNA microarray experiments were analyzed by average-linkage hierarchical clustering and significance analysis of microarrays (SAM). RESULTS: Eighteen tumor and non-tumor specimens were clearly divided by clustering analysis. Three features of gene expression profile were found in gastric adenocarcinoma and non-tumor tissues. The profile of differential gene expression in tumor and non-tumor tissues was mainly shown in feature B and feature C. In gastric adenocarcinoma tissues, the expression of genes in feature B was lower and that in feature C was higher. The profile of differential gene expression among gastric adenocarcinoma tissues was found in feature A. In feature A, the profile of similar gene expression was found in paired tumor and non-tumor tissues from 13 patients. SAM analysis showed that 19 genes in feature B and 12 genes in feature C were of significant difference between tumor and non-tumor specimens. The expression levels of genes related to cell cycle, growth factor, cell adhesion, and matrix remodeling were higher or lower in gastric adenocarcinoma tissues. CONCLUSION: Data from cDNA microarray experiments can clearly distinguish gastric adenocarcinoma from non-tumor tissues. The profiles show that gene expression in gastric adenocarcinomas is both homogeneous and heterogeneous. The homogeneous gene expression profile is found in both tumor and non-tumor tissues from 13 patients, suggesting that some gene aberrance is an early event of carcinogenesis of gastric adenocarcinoma. This study provides not only a new molecular basis for understanding biological properties of gastric adenocarcinoma, but also useful resources for future development of diagnostic and prognostic markers for gastric adenocarcinoma.
Assuntos
Adenocarcinoma/genética , Perfilação da Expressão Gênica , Análise de Sequência com Séries de Oligonucleotídeos , Neoplasias Gástricas/genética , Adulto , Idoso , Análise por Conglomerados , Feminino , Humanos , Masculino , Pessoa de Meia-IdadeRESUMO
Gastric neuroendocrine tumors (GNETs) are rare lesions characterized by hypergastrinemia that arise from enterochromaffin-like cells of the stomach. GNETs consist of a heterogeneous group of neoplasms comprising tumor types of varying pathogenesis, histomorphologic characteristics, and biological behavior. A classification system has been proposed that distinguishes four types of GNETs; the clinicopathological features of the tumor, its prognosis, and the patient's survival strictly depend on this classification. Thus, correct management of patients with GNETs can only be proposed when the tumor has been classified by an accurate pathological and clinical evaluation of the patient. Recently developed cancer therapies such as inhibition of angiogenesis or molecular targeting of growth factor receptors have been used to treat GNETs, but the only definitive therapy is the complete resection of the tumor. Here we review the literature on GNETs, and summarize the classification, clinicopathological features (especially prognosis), clinical presentations and current practice of management of GNETs. We also present the latest findings on new gene markers for GNETs, and discuss the effective drugs developed for the diagnosis, prognosis and treatment of GNETs.