RESUMEN
BACKGROUND: Hepatocellular carcinoma (HCC) is a serious threat to human lives and is usually diagnosed at the late stages. Recently, there has been a rapid advancement in the treatment options for HCC, but novel therapeutic targets are still needed, especially for precision medicine. AIMS: We aimed to investigate the involvement of non-coding RNA RP11-81H3.2 in HCC. METHODS: The expression of RP11-81H3.2 was examined in the blood samples of HCC patients, and in the human HCC cell lines, including HepG2, Smmc-7721, and Huh7. Cell proliferation was determined using the CCK-8 and EdU assay, and cell invasion and migration were determined using the transwell/wound healing assay. The effects of RP11-81H3.2 knockdown on in vivo tumor growth were evaluated utilizing the nude mice HepG2 tumor xenograft model. RESULTS: Here, we have identified a long non-coding RNA, RP11-81H3.2, which is enriched in HCC and can promote its proliferation, migration, and invasion both in vitro and in vivo. In addition, our results showed that RP11-81H3.2 binds to and regulate miR-490-3p expression in the HCC cells. Moreover, we found that RP11-81H3.2 regulates the expression of TNKS2 via miR-490-3p. Further, we found that RP11-81H3.2 and miR-490-3p form a regulatory loop; the release of RP11-81H3.2 leads to the suppression of miR-490-3p expression, thus, further enhancing the expression of RP11-81H3.2. CONCLUSIONS: Our data have provided a novel target for the diagnosis and treatment of HCC, and sheds light on the lncRNA-miRNA regulatory nexus that can control the HCC related pathogenesis.
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Carcinoma Hepatocelular/enzimología , Neoplasias Hepáticas/enzimología , MicroARNs/metabolismo , Oncogenes , ARN Largo no Codificante/metabolismo , Tanquirasas/biosíntesis , Animales , Carcinoma Hepatocelular/genética , Carcinoma Hepatocelular/patología , Movimiento Celular , Proliferación Celular , Regulación Neoplásica de la Expresión Génica , Células Hep G2 , Humanos , Neoplasias Hepáticas/genética , Neoplasias Hepáticas/patología , Ratones Desnudos , MicroARNs/genética , Invasividad Neoplásica , ARN Largo no Codificante/genética , Transducción de Señal , Tanquirasas/genética , Carga TumoralRESUMEN
BACKGROUND: Contactin1 (CNTN1) has been shown to play an important role in the invasion and metastasis of several tumors; however, the role of CNTN1 in breast cancer has not been fully studied. The purpose of this study is to investigate the role of CNTN1 in regulating tumor growth, migration and invasion in breast cancer. RESULTS: To investigate its function, CNTN1 was expressed in Hs578T cells. CNTN1 expression was confirmed by western blot, immunohistochemistry and real-time RT-PCR. The effect of CNTN1 overexpression on proliferation, migration and invasion of Hs578T breast cancer cells was assessed in vitro and in vivo. Our results showed that CNTN1 overexpression promoted Hs578T cell proliferation, cell cycle progression, colony formation, invasion and migration. Notably, overexpression of CNTN1 in Hs578T cells enhanced the growth of mouse xenograft tumors. CONCLUSIONS: CNTN1 promotes growth, metastasis and invasion of Hs578T breast cancer cell line. Thus, therapies targeting CNTN1 may prove efficacious for breast cancer. However, further investigation is required to understand the mechanism by which CNTN1 influences proliferation, metastasis and invasion in breast cancer.
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Neoplasias de la Mama/patología , Movimiento Celular , Contactina 1/metabolismo , Animales , Línea Celular Tumoral , Proliferación Celular , Femenino , Humanos , Masculino , Ratones Desnudos , Invasividad Neoplásica , Ensayos Antitumor por Modelo de XenoinjertoRESUMEN
BACKGROUND: Gastroesophageal reflux (GR) after radical resection of proximal gastric cancer (PGC) may influence survival; however, few studies have investigated survival in PGC patients who develop GR following radical resection. This study aimed to correlate the occurrence of GR after proximal gastrectomy (PG) and total gastrectomy (TG) with clinicopathological factors and long-term survival. METHODS: The PGC patient cohort was retrospectively grouped as follows: postoperative patients with and without GR (NGR). Clinicopathological characteristics and survival data were compared between the two groups. RESULTS: A total of 88 patients who underwent PG (53%) experienced postoperative GR; however, only 30 patients who underwent TG (14%) experienced GR (P = 0.000). The incidence of GR was significantly associated with surgical procedure (P < 0.01), tumor size (P < 0.01), infiltration depth (P < 0.01), lymph node metastasis (P = 0.018), postoperative distant metastasis (P < 0.01) and recurrence (P = 0.001). The 5-year overall survival of the GR group was significantly worse than that of the NGR group (39.3 vs. 46.5%, respectively; P = 0.046). The PG and TG groups had significantly different 5-year overall survival (45.2 vs. 50.9%, respectively; P = 0.047), and multivariate analysis revealed GR as an independent risk factor associated with poor overall survival. CONCLUSIONS: Patients who experienced GR after radical resection for PGC were more likely to develop recurrence and metastasis, leading to shorter survival. TG for PGC was associated with a more favorable 5-year overall survival than was PG. Thus, TG should be performed for PGC patients with tumors larger than 5 cm, T3/T4 disease or lymph node metastasis to improve their long-term survival.
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Gastrectomía/efectos adversos , Reflujo Gastroesofágico/etiología , Recurrencia Local de Neoplasia/epidemiología , Complicaciones Posoperatorias/etiología , Neoplasias Gástricas/mortalidad , Neoplasias Gástricas/patología , Anciano , Femenino , Reflujo Gastroesofágico/mortalidad , Reflujo Gastroesofágico/patología , Humanos , Masculino , Persona de Mediana Edad , Complicaciones Posoperatorias/mortalidad , Complicaciones Posoperatorias/patología , Estudios Retrospectivos , Neoplasias Gástricas/cirugía , Tasa de SupervivenciaRESUMEN
BACKGROUND Gastrointestinal stromal tumor (GIST) is an uncommon visceral sarcoma that arises predominantly in the gastrointestinal tract. Since GISTs are encountered infrequently and inflexible to traditional therapy, the aim of the present study was to explore the correlation of B-cell-specific Moloney murine leukemia virus insertion site 1 (BMI-1) mRNA and BMI-1 protein levels with the clinicopathological characteristics and prognosis significance of GISTs. MATERIAL AND METHODS GIST tissues and normal tissues were collected from 156 patients who had undergone surgical treatment. RT-qPCR and immunohistochemistry were used to measure the BMI-1 mRNA and protein levels in GIST tissues and normal tissues. Univariate survival analysis was used for determination of the factors that affect prognosis of GIST patients. Cox proportional hazards model was plotted to determine the independent risk factors for prognosis of GIST patients. RESULTS The BMI-1 mRNA and protein levels in GIST tissues were higher than those in normal tissues. BMI-1 mRNA and positive protein levels were correlated with the National Institutes of Health (NIH) risk grade, tumor diameter and infiltration, and metastasis. There was a short survival period for the patients with a positive protein level and a high mRNA level of BMI-1. The site of primary tumor, tumor diameter, NIH risk grade, infiltration, and metastasis, as well as BMI-1 mRNA and protein levels were independent risk factors for prognosis of GIST patients. CONCLUSIONS Taken together, these findings suggest there might be a relationship between BMI-1 mRNA and protein levels, and clinicopathological characteristics, including NIH risk grade, tumor size as well as infiltration and metastasis, of GIST patients. In addition, BMI-1 mRNA and protein levels were identified as independent risk factors for prognosis of GIST patients.
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Neoplasias Gastrointestinales/genética , Tumores del Estroma Gastrointestinal/genética , Complejo Represivo Polycomb 1/genética , Complejo Represivo Polycomb 1/metabolismo , Adulto , Anciano , Biomarcadores de Tumor/genética , Biomarcadores de Tumor/metabolismo , Supervivencia sin Enfermedad , Femenino , Neoplasias Gastrointestinales/metabolismo , Neoplasias Gastrointestinales/patología , Tumores del Estroma Gastrointestinal/metabolismo , Tumores del Estroma Gastrointestinal/patología , Predisposición Genética a la Enfermedad , Humanos , Inmunohistoquímica , Masculino , Persona de Mediana Edad , Factor de Transcripción PAX5/genética , Factor de Transcripción PAX5/metabolismo , Pronóstico , Modelos de Riesgos Proporcionales , ARN Mensajero/genética , ARN Mensajero/metabolismo , Factores de RiesgoRESUMEN
BACKGROUND: Analysis of aberrant hypermethylation in stool DNA might provide a novel strategy for noninvasive detection of colorectal cancer. AIMS: To explore the feasibility of detecting hypermethylation in Spastic paraplegia-20 promoter as a stool-based DNA marker for detection of colorectal cancer. METHODS: We collected 96 tissue and stool samples from patients with colorectal cancer and 30 stool samples healthy individuals. RESULTS: Hypermethylated Spastic paraplegia-20 occurs in 85.4% (82/96) of patients with colorectal cancer in the tissue samples. In the stool samples, the results indicate 80.2% (77/96) sensitivity and 100% (30/30) specificity of the test for detecting colorectal cancer by using the stool samples as a noninvasive method. CONCLUSIONS: The study reveals that hypermethylation in Spastic paraplegia-20 promoter is a highly specific and sensitive biomarker for screening colorectal cancer in stool samples as a noninvasive method.
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Neoplasias Colorrectales , ADN/genética , Paraplejía , Proteínas/genética , Anciano , Proteínas de Ciclo Celular , Neoplasias Colorrectales/complicaciones , Neoplasias Colorrectales/diagnóstico , Neoplasias Colorrectales/genética , Metilación de ADN/genética , Detección Precoz del Cáncer , Heces/química , Femenino , Humanos , Masculino , Persona de Mediana Edad , Paraplejía/complicaciones , Paraplejía/diagnóstico , Paraplejía/genética , Regiones Promotoras GenéticasRESUMEN
Rapidly proliferating cancer cells rely on increased glucose consumption for survival. The glucose analog 2-deoxy-D-glucose (2DG) cannot complete glycolysis and inhibits the growth of many types of cancers. It is unknown whether reduced glycolysis inhibits the growth of pancreatic cancer. Activation of nerve growth factor (NGF)-neurotrophic tyrosine kinase receptor type 1 (NTRK1) signaling leads to enhanced proliferation of these cells. We investigated the effect of 2DG treatment on the viability of NTRK1-transfected pancreatic cancer cells. After treatment with 2DG, the viability of pancreatic cancer cells was evaluated by MTT assay. SB203580 (a specific inhibitor of the p38-MAPK pathway) and PD98059 (an MAP2K1 [mitogen-activated protein kinase kinase 1, previously, MEK1] inhibitor) were used to inhibit p38-MAPK and ERKs, respectively. The percentage of apoptotic cells was determined by flow cytometry. Overexpression of NTRK1 in pancreatic cancer cells resulted in increased cell proliferation, which was reduced by PD98059-mediated inhibition of ERKs but not by suppression of p38-MAPK with SB203580. After treatment with 2DG, the percentage of apoptotic cells was greater in those with high expression of NTRK1 than in cells with low NTRK1 expression. Blocking the p38-MAPK pathway with SB203580 effectively abolished the apoptosis induced by 2DG. We conclude that pancreatic cancer cells with a high expression of NTRK1 are more sensitive to 2DG-induced apoptosis, through the p38-MAPK pathway.
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Desoxiglucosa/farmacología , Factor de Crecimiento Nervioso/metabolismo , Neoplasias Pancreáticas/metabolismo , Receptor trkA/metabolismo , Animales , Apoptosis/efectos de los fármacos , Línea Celular Tumoral , Proliferación Celular/efectos de los fármacos , Supervivencia Celular/efectos de los fármacos , Regulación Neoplásica de la Expresión Génica/efectos de los fármacos , Glucólisis , Humanos , Imidazoles/farmacología , Neoplasias Pancreáticas/patología , Piridinas/farmacología , Transducción de Señal/efectos de los fármacos , Proteínas Quinasas p38 Activadas por Mitógenos/antagonistas & inhibidores , Proteínas Quinasas p38 Activadas por Mitógenos/metabolismoRESUMEN
AIM: To identify discriminating protein patterns in serum samples among non-small cell lung cancer (NSCLC), chronic obstructive pulmonary disease (COPD), pneumonia, and healthy controls. To discover specific low molecular weight (LMW) serum peptidome biomarkers and establish a diagnostic pattern for NSCLCby using proteomic technology. METHODS: We used magnetic bead-based separation followed by matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) to identify patients with NSCLC, COPD, and pneumonia. A total of 154 serum samples were analyzed in this study, among which there were 60 serum samples from NSCLC patients, 30 from patients with other lung-related diseases (16 pneumonia patients and 14 patients with COPD) as disease controls, and 64 from healthy volunteers as healthy control. The mass spectra, analyzed using ClinProTools software, distinguished between cancer patients and healthy individuals based on GA algorithm model. RESULTS: In this study, we generated numerous discriminating m/z peaks as well as disease-specific discrimination peaks. A set of five potential biomarkers (m/z: 7,763.24, 1,012.61, 4,153.16, 1,450.55, and 2,878.89) could be used as the diagnostic biomarkers to distinguish NSCLCpatients from healthy controls. In the training set, patients with NSCLC could be identified with sensitivity of 97.5% and specificity of 98.8%. Similar results were obtained in the testing set, showing 80.7% sensitivity and 91.2% specificity. CONCLUSION: Our study demonstrated that a combined application of magnetic beads with MALDI-TOF MS technique was suitable for identification of serum biomarkers for NSCLC.
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Biomarcadores de Tumor/sangre , Carcinoma de Pulmón de Células no Pequeñas/sangre , Neoplasias Pulmonares/sangre , Péptidos/sangre , Algoritmos , Biomarcadores de Tumor/química , Estudios de Casos y Controles , Humanos , Peso Molecular , Péptidos/química , Neumonía/sangre , Proteoma/análisis , Proteoma/química , Proteómica , Enfermedad Pulmonar Obstructiva Crónica/sangre , Sensibilidad y Especificidad , Espectrometría de Masa por Láser de Matriz Asistida de Ionización DesorciónRESUMEN
OBJECTIVE: To elucidate the effect of proliferation and apoptosis induced by vitexicarpin in mutated p53 Hs578T cell line and study the expression of c-Myc, p21 and Bcl-2 protein in Hs578T and wild p53 MCF-7 cell pre-treated with vitexicarpin. METHODS: Cells were treated with various concentrations of vitexicarpin (0, 0.1, 0.2, 0.5, 1.0 micromol/L). MTT assays were used to detect cell proliferation at different time points with different doses of vitexicarpin. TUNEL assays were performed to examine apoptosis in cells pretreated with vitexicarpin. The authors detected three main proteins involved in apoptosis: c-Myc, bcl-2 and p21 protein in various concentrations of vitexicarpin-treated cells. To understand the function of c-Myc protein in the effect of vitexicarpin, the authors transiently transfected c-Myc protein in Hs578T cell and detected the cellular effect of vitexicarpin. RESULTS: Proliferation of Hs578T and MCF-7 cells were inhibited markedly by vitexicarpin at concentrations above 0.2 micromol/L (IC50 = 0.25 micromol/L and 0.53 micromol/L at 72 h respectively). TUNEL assays revealed that the rates of TUNEL positive cells were 10.15%, 27.33% and 35.34% when exposing Hs578T cells to 0.1, 0.2 and 0.5 micromol/L of vitexicarpin respectively. In control cells, the rates of TUNEL positive cells were 4.65%. Cells pretreated with higher concentrations of vitexicarpin expressed less c-Myc and Bcl-2 in Hs578T cells.In contrast, p21 decreased when cells were treated with the same conditions. When c-Myc transient transfection was performed in vitexicarpin-treated cells, the effect of p21 and Bcl-2 disappeared. The proliferative function of vitexicarpin declined in Hs578T/c-Myc cells. When treated with 0.5 micromol/L vitexicarpin, A value increased 1.53 times at 72 h. Conversely, A value decreased 48% at the same condition in MCF-7/c-Myc cells. CONCLUSION: The suppressing mechanism of vitexicarpin for malignant tumors is through c-Myc in p53 mutated Hs578T cells. And it is multi-directional and varies in different cells.
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Apoptosis/efectos de los fármacos , Neoplasias de la Mama/tratamiento farmacológico , Proliferación Celular/efectos de los fármacos , Flavonoides/farmacología , Apoptosis/genética , Neoplasias de la Mama/genética , Línea Celular Tumoral , Femenino , Humanos , Mutación , Proteína p53 Supresora de Tumor/genéticaRESUMEN
BACKGROUND: Alterations of mitochondrial DNA (mtDNA) have been implicated in carcinogenesis. We developed an accurate multiplex quantitative real-time PCR for synchronized determination of mtDNA and nuclear DNA (nDNA). We sought to investigate whether mtDNA content in the peripheral blood of breast cancer patients is associated with clinical and pathological parameters. METHODS: Peripheral blood samples were collected from 60 patients with breast cancer and 51 age-matched healthy individuals as control. DNA was extracted from peripheral blood for the quantification of mtDNA and nDNA, using a one-step multiplex real-time PCR. A FAM labeled MGB probe and primers were used to amplify the mtDNA sequence of the ATP 8 gene, and a VIC labeled MGB probe and primers were employed to amplify the glyceraldehyde-3-phosphate-dehydrogenase gene. mtDNA content was correlated with tumor stage, menstruation status, and age of patients as well as lymph node status and the expression of estrogen receptor (ER), progesterone receptor (PR) and Her-2/neu protein. RESULTS: The content of mtDNA in stage I breast cancer patients was significantly lower than in other stages (overall P = 0.023). Reduced mtDNA was found often in post menopausal cancer group (P = 0.024). No difference in mtDNA content, in regards to age (p = 0.564), lymph node involvement (p = 0.673), ER (p = 0.877), PR (p = 0.763), and Her-2/neu expression (p = 0.335), was observed. CONCLUSION: Early detection of breast cancer has proved difficult and current detection methods are inadequate. In the present study, decreased mtDNA content in the peripheral blood of patients with breast cancer was strongly associated with stage I. The use of mtDNA may have diagnostic value and further studies are required to validate it as a potential biomarker for early detection of breast cancer.
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Neoplasias de la Mama/sangre , Neoplasias de la Mama/genética , ADN Mitocondrial/sangre , Neoplasias de la Mama/patología , Femenino , Humanos , Persona de Mediana Edad , Estadificación de Neoplasias , Reacción en Cadena de la Polimerasa de Transcriptasa InversaRESUMEN
In this study, we aimed to explore the expression profiles of some known functional lncRNAs in esophageal adenocarcinoma (EAD) and to screening the potential prognostic makers, using data from The Cancer Genome Atlas (TCGA)-esophageal carcinoma (ESCA). Results showed that DLEU2 is a high potential OS related marker among 73 functional lncRNAs. DLEU2 and its intronic miR-15a and miR-16-1 expression were significantly upregulated in EAD compared with adjacent normal tissues. However, miR-15a and miR-16-1 expression were only weakly correlated with DLEU2 expression. Univariate and multivariate analysis confirmed that DLEU2 expression, but not miR-15a or miR-16-1 expression is an independent prognostic marker in terms of OS (HR:1.688, 95%CI: 1.085-2.627, p = 0.020) in EAD patients. The exon 9 of DLEU2 is very strongly co-expressed with DLEU2 (Pearson's r = 0.96) and showed better predictive value than total DLEU2 expression in predicting the OS of EAD patients. Multivariate analysis confirmed its independent prognostic value (HR:1.970, 95%CI: 1.266-3.067, p = 0.003), after adjustment of histologic grade, pathological stages and the presence of residual tumor. By checking the methylation status of DLEU2 gene, we excluded the possibility of the influence of two CpG sites near the DLEU2 exon 9 locus on its expression. In addition, although copy number alterations (CNAs) were observed DLEU2 gene, heterozygous loss (-1), low-level copy gain (+1) and high-level amplification (+2) had no significant association with DLEU2 transcription. Based on these findings, we infer that DLEU2 exon 9 expression might serve as a valuable biomarker of unfavorable OS in EAD patients.
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Adenocarcinoma/metabolismo , Biomarcadores de Tumor/biosíntesis , Neoplasias Esofágicas/metabolismo , Exones/genética , ARN Largo no Codificante/metabolismo , Proteínas Supresoras de Tumor/metabolismo , Adenocarcinoma/genética , Adenocarcinoma/mortalidad , Islas de CpG/genética , Epigénesis Genética , Neoplasias Esofágicas/genética , Neoplasias Esofágicas/mortalidad , Femenino , Humanos , Modelos Lineales , Masculino , Pronóstico , ARN Largo no Codificante/genética , Análisis de Supervivencia , Transferasas , Proteínas Supresoras de Tumor/genética , Regulación hacia ArribaRESUMEN
Gastric cancer (GC) is one of the leading causes of cancer-associated mortality worldwide. The aim of the present study was to investigate the mechanism of microRNA-4295 (miR-4295), which regulates cisplatin (DDP)-induced apoptosis in GC cells through the leucine-rich repeats and immunoglobulin-like domains 1 (LRIG1)-mediated epidermal growth factor receptor (EGFR)/phosphoinositide 3-kinase (PI3K)/protein kinase B (Akt) signaling pathway. Two cell lines were selected, one with the highest expression of miR-4295 and one with the lowest expression of LRIG1, for the experiments. The half maximal inhibitory concentration of DDP in the human GC MKN-28 and MKN-45 cell lines was calculated, and mitochondrial membrane potentials of the GC cells were detected by tetramethylrhodamine, ethyl ester, perchlorate staining. The proliferation and apoptosis of GC cells with or without DDP treatment were assessed by MTT assay and plate colony formation, as well as flow cytometry and TUNEL staining. Western blot analysis and reverse transcription-quantitative polymerase chain reaction were employed to determine the expression of EGFR/PI3K/Akt signaling pathway-related genes and apoptosis-related genes. LRIG1 was identified as a target gene of miR-4295. The expression of miR-4295 was upregulated, and the expression of LRIG1 was downregulated in GC cells. Furthermore, DDP enhanced the decrease in miR-4295 expression and the increase in LRIG1 expression in GC cells. miR-4295 promoted the proliferation and inhibited the DDP-induced apoptosis of GC cells without DDP treatment. In addition, miR-4295 increased the expression levels of EGFR, PI3K, Akt, p-PI3K and p-Akt, suggesting that miR-4295 promotes the activation of the EGFR/PI3K/Akt signaling pathway by targeting LRIG1. miR-4295 targeted and negatively regulated LRIG1 expression to activate the EGFR/PI3K/Akt signaling pathway, thereby promoting the proliferation of the GC cells and inhibiting the apoptosis of the GC cells induced by DDP. Therefore, miR-4295 may be a novel therapeutic target in patients with GC.
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Cisplatino/farmacología , Regulación hacia Abajo , Glicoproteínas de Membrana/genética , MicroARNs/genética , Transducción de Señal/efectos de los fármacos , Neoplasias Gástricas/genética , Regiones no Traducidas 3' , Línea Celular Tumoral , Proliferación Celular/efectos de los fármacos , Supervivencia Celular/efectos de los fármacos , Receptores ErbB/metabolismo , Regulación Neoplásica de la Expresión Génica/efectos de los fármacos , Humanos , Concentración 50 Inhibidora , Potencial de la Membrana Mitocondrial/efectos de los fármacos , Fosfatidilinositol 3-Quinasa/metabolismo , Proteínas Proto-Oncogénicas c-akt/metabolismo , Neoplasias Gástricas/tratamiento farmacológicoRESUMEN
OBJECTIVE: To determine the clinical application of the new classification of adenocarcinoma of esophagogastric junction (AEG). METHODS: The data of cancer of distal esophagus, cancer of cardia, and proximal gastric cancer were reviewed. Clinicopathologic characteristics, surgical modes and survival were analyzed according to Siewert's standards. RESULTS: Among the 203 patients that were up to the standard, 29 had adenocarcinoma of the distal esophagus (Type I), 80 had true carcinoma of cardia (Type II), and 94 had subcardial carcinoma (Type III). The 5-year survival rates of the 3 types of patients after the operation were 34% for Type I, 27.5% for Type II, and 24.5% for Type III (P<0.05). Further analysis of the patients with curative resection suggested there was no significant difference in the 5-year survival rates, with 37.5% for Type I, 34.5% for Type II, and 33.3% for Type III (P>0.05). CONCLUSION: Difference has been found in the clinicopathologic characteristics of the 3 types of adenocarcinoma of the esophagogastric junction. The exact relation of the 3 types is still unknown. The TNM classification, complete tumor resection and the extent of lymph node metastasis are critical for the prognosis of the patients.
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Adenocarcinoma/clasificación , Neoplasias Esofágicas/clasificación , Unión Esofagogástrica/patología , Adenocarcinoma/mortalidad , Adenocarcinoma/cirugía , China , Neoplasias Esofágicas/mortalidad , Neoplasias Esofágicas/cirugía , Esofagectomía , Unión Esofagogástrica/cirugía , Femenino , Humanos , Masculino , Estudios Retrospectivos , Análisis de Supervivencia , Tasa de SupervivenciaRESUMEN
PURPOSE: A quantitative multiple-marker reverse transcriptase (RT)-polymerase chain reaction (PCR) assay for sensitive detection of cancer cells in axillary drainage fluid was developed to examine whether the presence of cancer cells in axillary drainage fluid can be used as a predictor of locoregional recurrence (LRR) in patients with breast cancer who had T1/2 primary tumors and one to three positive axillary lymph nodes treated with modified radical mastectomy without adjuvant radiotherapy. METHODS AND MATERIALS: Axillary drainage fluid was collected from 126 patients with invasive ductal carcinoma of the breast who were treated with modified radical mastectomy and were found to have one to three positive axillary nodes. Cancer cells in axillary drainage fluid were detected by RT-PCR assay using primers specific for carcinoembryonic antigen (CEA) and cytokeratin-19 (CK-19) together with numerous clinicopathologic and treatment-related factors and were analyzed for their impact on LRR. RESULTS: A total of 38 patients suffered LRR during follow-up and the multimarker RT-PCR assays for CEA and CK-19 in the axillary drainage fluid both were positive in 34 patients (27.0%), of which 29 patients had LRR. In univariate analysis, the 5-year LRR-free survival showed higher rates in patients with PCR-negative findings in axillary drainage fluid (p<0.0001), age>or=40 years old (p<0.0001), tumor size<2.5 cm (p<0.0001), negative lymph-vascular space invasion (p=0.026), and T1 status (<0.0001); in multivariate analysis, PCR-positive findings together with age and tumor size were found to be independent predictors of LRR (all p<0.05). CONCLUSION: Multiplex RT-PCR assay for CEA and CK-19 was highly sensitive for detection and might be useful for prediction of LRR in such subgroup breast cancer patients.
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Biomarcadores de Tumor/análisis , Neoplasias de la Mama/patología , Antígeno Carcinoembrionario/análisis , Carcinoma Ductal de Mama/secundario , Queratinas/análisis , Ganglios Linfáticos/metabolismo , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa/métodos , Adulto , Análisis de Varianza , Axila , Neoplasias de la Mama/mortalidad , Neoplasias de la Mama/cirugía , Carcinoma Ductal de Mama/mortalidad , Carcinoma Ductal de Mama/cirugía , Femenino , Humanos , Ganglios Linfáticos/patología , Metástasis Linfática , Mastectomía Radical Modificada , Persona de Mediana Edad , Modelos Biológicos , Recurrencia Local de Neoplasia/metabolismo , Recurrencia Local de Neoplasia/patología , Neoplasia Residual , Radioterapia Adyuvante , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa/normas , Sensibilidad y EspecificidadRESUMEN
MicroRNAs (miRNAs) are small noncoding RNAs that function in diverse biological processes. However, little is known about the precise role of microRNAs in the functioning of airway smooth muscle cells (ASMCs). Here, we investigated the potential role and mechanisms of the miR-143 -3p on proliferation and the extracellular matrix (ECM) protein production of ASMCs. We demonstrated that miR-143-3p was aberrantly lower in ASMCs isolated from individuals with asthma than in individuals without asthma. Meanwhile, TGF-ß1 caused a marked decrease in a time-dependent manner in miR-143-3p expression in ASMCs from asthmatics. Additionally, the overexpression of miR- 143-3p robustly reduced TGF-ß1-induced ASMCs proliferation and downregulated CDK and cyclin expression, whereas the inhibition of miR-143-3p significantly enhanced ASMCs proliferation and upregulated the level of CDKs and cyclins. Re-expression of miR-143-3p attenuated ECM protein deposition reflected as a marked decrease in the expression of type I collagen and fibronectin, whereas miR-143-3p downregulation caused an opposite effect on the expression of type I collagen and fibronectin. Moreover, qRT-PCR and western blot analysis indicated that miR-143-3p negatively regulated the expression of nuclear factor of activated T cells 1 (NFATc1). Subsequent analyses demonstrated that NFATc1 was a direct and functional target of miR-143-3p, which was validated by the dual luciferase reporter assay. Most importantly, the overexpression of NFATc1 effectively reversed the inhibition of miR-143-3p on TGF-ß1-induced proliferation, and strikingly abrogated the effect of miR-143-3p on the expression of CDK4 and Cyclin D1. Together, miR-143-3p may function as an inhibitor of asthma airway remodeling by suppressing proliferation and ECM protein deposition in TGF-ß1-mediated ASMCs via the negative regulation of NFATc1 signaling, suggesting miR-143-3p as a potential therapeutic target for asthma.
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Remodelación de las Vías Aéreas (Respiratorias)/inmunología , Asma/inmunología , Regulación de la Expresión Génica/inmunología , MicroARNs/inmunología , Factores de Transcripción NFATC/biosíntesis , Adulto , Asma/genética , Asma/patología , Western Blotting , Bronquios/inmunología , Bronquios/metabolismo , Bronquios/patología , Proliferación Celular/fisiología , Células Cultivadas , Regulación hacia Abajo , Ensayo de Inmunoadsorción Enzimática , Matriz Extracelular/patología , Femenino , Humanos , Masculino , MicroARNs/metabolismo , Músculo Liso/inmunología , Músculo Liso/patología , Factores de Transcripción NFATC/inmunología , Reacción en Cadena en Tiempo Real de la Polimerasa , Factor de Crecimiento Transformador beta1/inmunología , Factor de Crecimiento Transformador beta1/metabolismoRESUMEN
AIM: We examined the methylation status of SNCA and FBN1 genes in patients' paired tissue and stool samples for detection of colorectal cancer (CRC). PATIENTS AND METHODS: 89 DNA tissue samples (normal/cancer) and corresponding stool samples were analyzed in our study. In addition, 30 stool samples were collected as healthy controls. RESULTS: The methylation level of those samples was measured by methylation-specific polymerase chain reaction (MSP). The result shows that compared with the paired controls, both SNCA and FBN1 were significantly hypermethylated in CRC patients in tissue samples (P < 0.001). In the stool samples, hypermethylated SNCA and FBN1 were detected to be significantly higher than that in normal stool samples (P < 0.001). The combined sensitivity of at least one positive among the two markers in stool samples was 84.3%, with a specificity of 93.3%. In addition, our experiment suggested that the positive rates of SNCA and FBN1 in Dukes A stage were significantly higher than that of FOBT (P = 0.039; P = 0.006, resp.). CONCLUSION: We concluded that methylation testing of SNCA and FBN1 genes in stool sample may offer a good alternative in a simple, promising, and noninvasive detection of colorectal cancer.
Asunto(s)
Neoplasias Colorrectales/genética , Metilación de ADN , Detección Precoz del Cáncer/métodos , Heces/química , Proteínas de Microfilamentos/genética , alfa-Sinucleína/genética , Adulto , Anciano , Estudios de Casos y Controles , Neoplasias Colorrectales/patología , Femenino , Fibrilina-1 , Fibrilinas , Humanos , Masculino , Persona de Mediana EdadRESUMEN
OBJECTIVE: To study the clinical significance and effect of p21, p53 protein as well as proliferating cell nuclear antigen (PCNA) on the occurrence and development of pancreatic carcinoma. METHOD: p21, p53 protein and PCNA expressions were detected in specimens from 30 patients with pancreatic carcinoma and 3 samples of normal pancreatic tissue by immunohistochemistry. The data were analyzed together with clinical findings. RESULTS: The positive expression rates of p21 and p53 proteins were 75.0% and 57.3% respectively in pancreatic carcinoma, which were significantly different from those in the normal tissue (P<0.05). p21 and p53 proteins were positively correlated (P<0.05). The positive expression of PCNA was 43.33%+/-17.99%, that was significantly higher than that in the normal pancreatic tissue (P<0.05). The expression of PCNA was correlated with the histological grade (P<0.05). The positive expression rate was consistent with the exacerbation of cancer. The expression was also correlated significantly with prognosis and p53 expression (P<0.05). CONCLUSIONS: The occurrence and development of pancreatic cancer are the result of associated function for many oncogenes and antioncogenes. PCNA may be helpful to identify malignant degree and prognosis of pancreatic cancer.
Asunto(s)
Neoplasias Pancreáticas/metabolismo , Antígeno Nuclear de Célula en Proliferación/análisis , Proteínas Proto-Oncogénicas p21(ras)/metabolismo , Proteína p53 Supresora de Tumor/metabolismo , Estudios de Casos y Controles , Humanos , Inmunohistoquímica , Neoplasias Pancreáticas/patología , Antígeno Nuclear de Célula en Proliferación/metabolismo , Distribución TisularRESUMEN
Explore potential screening biomarkers for noninvasive diagnosis of colorectal cancer (CRC) by testing methylation of the miR-34a and miR-34b/c promoter in CRC patients' tissue and stool samples. Methylation-specific PCR analyses were performed on sample DNAs: 82 pairs of normal/cancer samples, 82 CRC patients' stool samples, 40 healthy volunteer stool samples, and 20 healthy volunteer blood samples were recruited. miR-34a has been found methylated in 65 of 82 (79.3%) the CRC tissue samples, but only 36 of 82 (43.9%) in corresponding normal samples. And when testing miR-34a in stool, 63 of 82 (76.8 %) CRC stool samples were observed methylated, and 2 of 40 (5%) healthy samples were observed methylated. The methylation for miR-34b/c has been found in 80 of 82 (97.5%) CRC tissue samples, 49 of 82 (59.8%) corresponding CRC normal samples, and 74 of 79 (93.6%) CRC stool samples. Yet we did not detect any methylation from healthy volunteers stool samples or healthy adult blood samples. Results indicated 76.8 % sensitivity and 93.6% specificity of the miR-34a methylation test for detecting CRC using stool samples. Meanwhile, the sensitivity and specificity of miR-34b/c were 95 and 100%, respectively. Moreover, our results revealed that abnormal DNA methylation of miR-34a was correlated with lymph metastasis (P = 0.010). Abnormal methylation of miR-34a and miR-34b/c genes might be regarded as potential biomarkers for noninvasive screening and diagnosis of colorectal cancer.
Asunto(s)
Biomarcadores de Tumor/metabolismo , Neoplasias Colorrectales/genética , Neoplasias Colorrectales/metabolismo , Regulación Neoplásica de la Expresión Génica , MicroARNs/metabolismo , Adulto , Anciano , Anciano de 80 o más Años , Metilación de ADN , Heces , Femenino , Voluntarios Sanos , Humanos , Metástasis Linfática , Masculino , Persona de Mediana Edad , Reacción en Cadena de la Polimerasa , Sensibilidad y Especificidad , Análisis de Secuencia de ADNRESUMEN
This study aimed to investigate the involvement of c-erbB-2, encoded by the receptor tyrosine kinase ERBB2 gene, in the pathogenesis of colorectal cancer and to validate its potential as an anticancer target. Immunohistochemical and histopathological analyses were applied in tissue samples derived from 80 colorectal cancer patients. ERBB2 stable small hairpin RNA (shRNA) knockdown in HT29 human colorectal cancer cells was confirmed by RT-PCR and western blotting. Cell cycle profile and apoptosis were measured using PI or Annexin V-PI dual staining. A significant correlation between ERBB2 levels and Dukes' stage of colorectal cancer, in both the primary malignancy and lymph node metastatic tissues, was observed. ERBB2-depleted HT-29 cells exhibited increased sensitivity to radiation compared to control cells, likely due to enhanced G0/G1 phase cell cycle arrest and apoptosis. ERBB2 may be involved in the malignancy and metastasis of colorectal cancer. Overexpressed ERBB2 may constitute a potential target for colorectal cancer therapy.
Asunto(s)
Neoplasias Colorrectales/patología , Interferencia de ARN , Tolerancia a Radiación/genética , Receptor ErbB-2/genética , Adulto , Anciano , Apoptosis/efectos de la radiación , Línea Celular Tumoral , Neoplasias Colorrectales/metabolismo , Femenino , Puntos de Control de la Fase G1 del Ciclo Celular/efectos de la radiación , Técnicas de Silenciamiento del Gen , Humanos , Inmunohistoquímica , Metástasis Linfática , Masculino , Persona de Mediana Edad , Estadificación de Neoplasias , ARN Interferente Pequeño/metabolismo , Receptor ErbB-2/antagonistas & inhibidores , Receptor ErbB-2/metabolismoRESUMEN
AIM: To study the changes of the cell growth, cell cycle distribution and cell apoptosis of colon cancer cell, HT-29, when C-erbB2 gene was knockdown by shRNA against C-erbB2. METHODS: Cell growth, cell cycle distribution and cell apoptosis were compared among three groups including plasmid experimental group(PEG), transfected reagent control group(TRCG)and negative plasmid control group(NPCG). Cell growth was measured by MTT assay. Cell cycle distribution and cell apoptosis were detected by flow cytometry. RESULTS: The inhibition rate of cell growth of PEG, TRCG and NPCG were 39.65%, 7.23% and 8.05% respectively. The cell growth was significantly inhibited in PEG(P<0.01). The cells of G0/G1 phase were 74.93%, 67.19%, 68.05% respectively in PEG, TRCG and NPCG. The cells of G0/G1 phase in PEG were significantly more than those in TRCG and NPCG(P<0.05).While the cells of S phase were 7.81%, 14.02%, 13.70% in PEG, TRCG and NPCG respectively. The cells of S phase in PEG were significantly less than those in TRCG or NPCG(P<0.05).The cell apoptosis rate were 19.21%, 3.13%, 4.08% in PEG, TRCG and NPCG respectively. The cell apoptosis rate in PEG was significantly higher than those in TRCG or NPCG(P<0.01). CONCLUSION: Cell growth was inhibited by shRNA against C-erbB2 gene. Cell cycle was blocked in G0/G1 phase and apoptosis was induced by C-erbB2 shRNA. This indicates C-erbB2 gene plays important roles in the carcinogenesis and development of colon cancer.