RESUMEN
The increasing expression of microRNA155 (miR155) and decreasing expression of RNAbinding protein quaking (QKI) in colon cells have been observed previously. In this study, we attempted to establish the correlation between miR155 and QKI. In addition, we assessed whether the expression of miR155 and QKI is linked to the proliferation and invasion capabilities of colon cells. Firstly, nineteen tumor samples, divided into two groups according to the presence or absence of lymphatic metastasis, were obtained from colon cancer patients at the First Affiliated Hospital of Wenzhou Medical University, China. The expression level of miR155 and QKI was measured by quantitative polymerase chain reaction (qPCR). Secondly, the GES1, SW480 and COLO205 cell lines were cultured and the expression level of QKI and miR155 was also assessed by qPCR. Thirdly, a luciferase reporter gene assay was performed to detect the association between miR155 and QKI, and qPCR and western blot analysis were performed to confirm the effects of miR155 on the expression of QKI at the mRNA and protein level. Subsequently, the SW480 cells were used in the following experiments. Following treatment with miR155 inhibitor and QKI overexpression vector, western blot analysis, propidium iodide (PI) staining and a cell scratch assay were carried out to assess the effects of miR155 on the proliferation and invasion potential of colon cancer cells. qPCR findings revealed higher miR155 expression and lower QKI expression in colon cancer tissues as well as the colon cancer cell lines SW480 and COLO205. The relative luciferase activity of the 3' untranslated region (3'UTR) was decreased by approximately 45% when SW480 cells stimulated by mimicmiR155 were combined with the wildtype 3'UTR constructs. In addition, when the cells were treated with mimicmiR155, QKI expression was significantly decreased at the mRNA and protein level. These outcomes revealed that miR155 decreased the production of QKI by acting on the 3'UTR of the QKI gene. Furthermore, PI staining and the cell scratch assay revealed that miR155 influenced the cell cycle and invasion abilities of colon cancer cells by directly targeting QKI and decreased the production of QKI by acting on the 3'UTR of the QKI gene. This study has demonstrated the correlation between miR155 and QKI, in which miR155 regulates the cell cycle and invasion ability of colon cancer cells via the modulation of QKI expression. Our study provides novel therapeutic strategies for colon cancer therapy.
Asunto(s)
Neoplasias del Colon/genética , MicroARNs/genética , Interferencia de ARN , ARN Mensajero/genética , Proteínas de Unión al ARN/genética , Regiones no Traducidas 3' , Secuencia de Bases , Sitios de Unión , Cadherinas/genética , Cadherinas/metabolismo , Ciclo Celular/genética , Línea Celular Tumoral , Movimiento Celular/genética , Proliferación Celular , Neoplasias del Colon/metabolismo , Neoplasias del Colon/patología , Expresión Génica , Humanos , Lactasa/genética , Lactasa/metabolismo , MicroARNs/química , MicroARNs/metabolismo , ARN Mensajero/química , Proteínas de Unión al ARN/metabolismoRESUMEN
AIM: To study any correlation of LKB1 expression with prognosis in hepatocellular carcinoma (HCC) cases. METHODS: A total of 70 HCC patients and 20 primary intrahepatic stone patients in the first affiliated hospital of Wenzhou Medical College were enrolled in this study. LKB1 expression was detected by immunohistochemistry. Patients were followed-up and prognostic factors were evaluated. RESULT: LKB1 expression was decreased in the HCC samples. Loss of LKB1 expression in HCC was significantly related to histologic grade (P=0.010), vascular invasion (P=0.025) and TMN stage (P=0.011). Patients showing negative LKB1 expression had a significantly shorter disease-free and overall survival than those with positive expression (P = 0.001, P=0.000, respectively). Multivariate Cox regression analysis indicated that LKB1 expression level was an independent factor of survival (P = 0.033). CONCLUSION: HCC patients with decreased expression LKB1 have a poor prognosis. The loss of LKB1 expression is correlated with a lower survival rate.
Asunto(s)
Biomarcadores de Tumor/metabolismo , Carcinoma Hepatocelular/mortalidad , Neoplasias Hepáticas/mortalidad , Hígado/metabolismo , Proteínas Serina-Treonina Quinasas/metabolismo , Quinasas de la Proteína-Quinasa Activada por el AMP , Adulto , Anciano , Carcinoma Hepatocelular/metabolismo , Carcinoma Hepatocelular/patología , Carcinoma Hepatocelular/cirugía , Femenino , Estudios de Seguimiento , Humanos , Técnicas para Inmunoenzimas , Hígado/patología , Neoplasias Hepáticas/metabolismo , Neoplasias Hepáticas/patología , Neoplasias Hepáticas/cirugía , Masculino , Persona de Mediana Edad , Invasividad Neoplásica , Estadificación de Neoplasias , Pronóstico , Tasa de SupervivenciaAsunto(s)
Adenocarcinoma/cirugía , Colecistectomía/métodos , Neoplasias de la Vesícula Biliar/cirugía , Adenocarcinoma/diagnóstico , Adenocarcinoma/patología , Adenocarcinoma Mucinoso/diagnóstico , Adenocarcinoma Mucinoso/patología , Adenocarcinoma Mucinoso/cirugía , Adulto , Anciano , Carcinoma de Células Escamosas/diagnóstico , Carcinoma de Células Escamosas/patología , Carcinoma de Células Escamosas/cirugía , Femenino , Estudios de Seguimiento , Neoplasias de la Vesícula Biliar/diagnóstico , Neoplasias de la Vesícula Biliar/patología , Humanos , Masculino , Persona de Mediana Edad , Estadificación de Neoplasias , Estudios Retrospectivos , Tasa de SupervivenciaRESUMEN
Recently, two hepatic lineage markers epithelial cell adhesion molecule (EpCAM) and α-fetoprotein (AFP) were used to classify hepatocellular carcinoma (HCC) into four subtypes with prognostic implication. In the present study, we further evaluated the clinicopathologic and angiogenic characteristics among these HCC subtypes. EpCAM expression was investigated by immunohistochemistry in 115 HCC primary tumors. Based on EpCAM immunostaining and serum AFP levels, 115 HCC cases were classified into four subtypes: EpCAM+AFP+ (26.1%), EpCAM-AFP+ (20.0%), EpCAM+AFP- (20.8%), and EpCAM-AFP- (33.1%). EpCAM+AFP+ and EpCAM-AFP+ HCC were associated with late TNM stages and high frequencies of venous invasion, whereas EpCAM+AFP- and EpCAM-AFP- subtypes were associated with early TNM stages and low frequencies of venous invasion. Furthermore, EpCAM+AFP+ HCC had a significantly higher microvessel density (MVD) and higher level of VEGF (Vascular epithelial growth factor) expression than the other three subtypes. In conclusion, our study indicated that subtype classification of HCC based on EpCAM and AFP status had clinicopathologic and biologic implications in aggressive phenotype and angiogenesis. We also suggest that the EpCAM+AFP+ HCC patients might be potential therapeutic candidates for anti-angiogenesis therapy.
Asunto(s)
Carcinoma Hepatocelular/clasificación , Carcinoma Hepatocelular/patología , Neoplasias Hepáticas/clasificación , Neoplasias Hepáticas/patología , Neovascularización Patológica/patología , Anciano , Antígenos de Neoplasias/biosíntesis , Biomarcadores de Tumor/análisis , Carcinoma Hepatocelular/metabolismo , Moléculas de Adhesión Celular/biosíntesis , Molécula de Adhesión Celular Epitelial , Femenino , Humanos , Inmunohistoquímica , Neoplasias Hepáticas/metabolismo , Masculino , Persona de Mediana Edad , Estadificación de Neoplasias , Neovascularización Patológica/clasificación , Neovascularización Patológica/metabolismo , alfa-Fetoproteínas/biosíntesisRESUMEN
AIM: To investigate the effects and possible mechanisms of tanshinone II-A, an alcohol extract of the root of Salvia miltiorrhiza Bunge, on tumor invasion and metastasis of human colon carcinoma (CRC) cells. METHODS: The effects of tanshinone II-A on invasion and metastasis of CRC cell lines HT29 and SW480 were evaluated by in vitro and in vivo assays. Western blotting was used to investigate possible molecular mechanisms of tanshinone II-A anti-cancer actions. RESULTS: Tanshinone II-A inhibited migration and invasion of CRC cells in a dose-dependent manner. The inhibitory effect also depended on time, with the most significant effects observed at 72 h. Tanshinone II-A also significantly inhibited in vivo metastasis of colon carcinoma SW480 cells. It inhibited in vitro and in vivo invasion and metastasis of CRC cells by reducing levels of urokinase plasminogen activator (uPA) and matrix metalloproteinases (MMP)-2 and MMP-9, and by increasing levels of tissue inhibitor of matrix metalloproteinase protein (TIMP)-1 and TIMP-2. Tanshinone II-A was also shown to suppress the nuclear factor-kappaB (NF-kappaB) signal. CONCLUSION: Tanshinone II-A inhibited in vitro and in vivo invasion and metastasis of CRC cells. The effect resulted from changes in the levels of uPA, MMP-2, MMP-9, TIMP-1 and TIMP-2, and apparent inhibition of the NF-kappaB signal transduction pathway.
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Antineoplásicos Fitogénicos/farmacología , Neoplasias del Colon/tratamiento farmacológico , Neoplasias Colorrectales/tratamiento farmacológico , Fenantrenos/farmacología , Abietanos , Animales , Antineoplásicos Fitogénicos/administración & dosificación , Antineoplásicos Fitogénicos/aislamiento & purificación , Línea Celular Tumoral , Neoplasias del Colon/patología , Neoplasias Colorrectales/patología , Relación Dosis-Respuesta a Droga , Células HT29 , Humanos , Ratones , Ratones Desnudos , Invasividad Neoplásica/prevención & control , Metástasis de la Neoplasia/prevención & control , Fenantrenos/administración & dosificación , Fenantrenos/aislamiento & purificación , Extractos Vegetales/administración & dosificación , Extractos Vegetales/farmacología , Raíces de Plantas , Salvia miltiorrhiza/química , Transducción de Señal/efectos de los fármacos , Factores de TiempoRESUMEN
OBJECTIVE: To analyze the expression of DD3 mRNA in the prostate tissues. METHODS: DD3 mRNA was detected by realtime fluorescent quantitative reverse transcription polymerase chain reaction (FQ-RT-PCR) based on the Taqman technique in the tissues of 27 patients with non-prostate cancer( NPCa), 21 prostate cancer( PCa), 39 benign prostatic hyperplasia (BPH) and 15 normal prostate (NP). The ROC curve was used to evaluate the diagnostic value of DD3 mRNA. RESULTS: DD3 mRNA expression was not detected in the NPCa tissues. The median expressions of DD3 mRNA in PCa, BPH and NP tissues were 7. 2 x 10(6), 2. 5 x 10(4) and 1.5 x 10(4) copies/mg tissue, respectively. The DD3 mRNA expression levels were significantly different between nonmalignant and malignant tissues (P < 0.01). No significant differences in DD3 mRNA expression were detected between the NP and BPH tissues and no significant correlation was found between the DD3 mRNA expression and clinical pathological parameters. The AUC-ROC was 0.937 (95% CI: 0.879 - 0.995) at cutoff value 1.4 x 10(5) copies/mg tissue. The sensitivity, specificity, accuracy, positive predictive value, negative predictive value, positive likelihood ratio and negative likelihood ratio for DD3 were 90.5%, 85.0%, 86.7%, 76.0%, 94.3%, 6.03 and 0.11 respectively. CONCLUSION: The DD3 mRNA expression is confined to prostate tissues and highly upregulated in PCa tissues. It has a potential application value in the early diagnosis of prostate cancer and the follow-up of the patient.
Asunto(s)
Antígenos de Neoplasias/biosíntesis , Neoplasias de la Próstata/metabolismo , Anciano , Anciano de 80 o más Años , Antígenos de Neoplasias/genética , Humanos , Masculino , Persona de Mediana Edad , Estadificación de Neoplasias , Hiperplasia Prostática/metabolismo , Hiperplasia Prostática/patología , Neoplasias de la Próstata/patología , ARN Mensajero/genética , Curva ROC , Reacción en Cadena de la Polimerasa de Transcriptasa InversaRESUMEN
AIM: To detect the DNA binding activity of nuclear factor-kappaB (NF-kappaB) in rat hepatocyte and to investigate the effects of NF-kappaB on rat hepatocyte regeneration and apoptosis after 70% portal branch ligation. METHODS: Sixty Wistar rats were randomly divided into control group and portal branch ligation group. The animals were killed 12 h, 1, 2, 3, 7, and 14 d after surgery to determine the contents of plasma ALT. Hepatocytes were isolated and nuclear protein was extracted. DNA binding activity of NF-kappaB was measured by EMSA. Hepatocyte regeneration and apoptosis were observed under microscope by TUNEL staining. The ultrastructural changes of liver were observed under electron microscope. RESULTS: Seventy percent portal branch ligation produced atrophy of the ligated lobes and the perfused lobes underwent compensatory regeneration, the total liver weight and plasma ALT levels were maintained at the level of sham-operated animals throughout the experiment. After 2 d of portal branch ligation, DNA binding activity of NF-kappaB in hepatocyte increased and reached its peak, the number of apoptotic hepatocyte in the ligated lobes and the number of mitotic hepatocyte in the perfused lobes also reached their peak. Typical apoptotic changes and evident fibrotic changes in the ligated lobes were observed under electron microscope. CONCLUSION: After 70% portal branch ligation, DNA binding activity of NF-kappaB in hepatocyte is significantly increased and NF-kappaB plays an important role in hepatocyte regeneration and apoptosis.