RESUMO
BACKGROUND: Cortactin is an important regulator involved in invasion and migration of hepatocellular carcinoma (HCC). The aim of this study was to elucidate the forecasting role of cortactin in resectable HCCs. METHODS: We compared the invasiveness and motility among liver epithelial cell line and HCC cell lines by using Transwell assay and wound healing assay. We further investigated the CTTN mRNA expression by real-time PCR. Next, 91 HCC and 20 normal liver tissue samples were detected by IHC and real-time PCR. Finally, we analyzed the clinicopathologic features and survival time of the HCC cases. RESULTS: We identified that HepG2, LM3, and SK-Hep-1 had more invasiveness and motility (P <0.05). Compared with liver epithelial cell line, CTTN expression was higher in LM3, HepG2, and MHCC97-L (P <0.01) and lower in SK-Hep-1 (P <0.05). IHC examination showed cortactin expression was closely relative to TNM stage (AJCC/UICC), cancer embolus, and metastasis (P <0.01). Cortactin overexpression indicated a longer survival time of 52 ± 8.62 months and low expression of a shorter survival time of 20 ± 4.95 months (P <0.01). Cortactin examination has more predictive power in patients with Child-Pugh grade A and BCLC stage 0-B. CONCLUSIONS: Overexpression of cortactin is closely associated with poor human HCCs prognosis that caused by cancer embolus and metastasis. Cortactin and CTTN should be used for differentiating varieties of survival for patients after HCC resection.
Assuntos
Biomarcadores Tumorais/metabolismo , Carcinoma Hepatocelular/mortalidade , Cortactina/metabolismo , Neoplasias Hepáticas/mortalidade , Fígado/metabolismo , Biomarcadores Tumorais/genética , Carcinoma Hepatocelular/metabolismo , Carcinoma Hepatocelular/secundário , Linhagem Celular Tumoral , Movimento Celular , Cortactina/genética , Feminino , Seguimentos , Humanos , Fígado/patologia , Neoplasias Hepáticas/metabolismo , Neoplasias Hepáticas/patologia , Masculino , Pessoa de Meia-Idade , Gradação de Tumores , Invasividade Neoplásica , Estadiamento de Neoplasias , Prognóstico , RNA Mensageiro/genética , Reação em Cadeia da Polimerase em Tempo Real , Estudos Retrospectivos , Taxa de Sobrevida , CicatrizaçãoRESUMO
The major immediate early (MIE) gene of cytomegalovirus plays a key role in determining the activation and replication of cytomegalovirus, which represents the most important event signaling the onset of virus-induced disease relapse. The viral-encoded chemokine receptor homolog US28 can constitutively activate many cellular transcription factors, which can bind to the promoter/enhancer of the MIE gene and activate its transcription. Using reporter gene assays in HEK293 cells, we found that US28 enhanced the transcription efficiency of MIE and other genes via cAMP response element-binding protein (CREB). Inhibition of CREB partially blocked the effect of US28, whereas forskolin enhanced this effect. There was a direct correlation between CREB and transcription of MIE gene. These data, together with the broad-spectrum effect of cellular transcription factors, suggest that US28 may be involved in the very early transcription of the host cell during virus activation.
Assuntos
Proteína de Ligação ao Elemento de Resposta ao AMP Cíclico/metabolismo , Elementos Facilitadores Genéticos/genética , Regulação Viral da Expressão Gênica , Genes Precoces/genética , Regiões Promotoras Genéticas/genética , Receptores de Quimiocinas/fisiologia , Proteínas Virais/fisiologia , Western Blotting , Proteína de Ligação ao Elemento de Resposta ao AMP Cíclico/genética , Humanos , Luciferases/metabolismo , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Fatores de Transcrição/metabolismo , Transcrição GênicaRESUMO
OBJECTIVE: To investigate the effects of secondary left-sided portal hypertension (LSPH) on the radical operation rate of patients with pancreatic cancer and systemically evaluate the prognosis of patients with LSPH secondary to pancreatic cancer after radical surgery. MATERIALS AND METHODS: The data of patients with pancreatic cancer who underwent laparotomy over a 15-year period in Department of Hepatobiliary Surgery of Chinese PLA Air Force General Hospital from Jan. 1, 1997, to Jun. 30, 2012 was retrospectively reviewed. RESULTS: A total of 362 patients with pancreatic cancer after laparotomy were selected, including 73 with LSPH and 289 without LSPH. Thirty-five patients with LSPH (47.9%) and 147 without non-LSPH (50.9%) respectively underwent radical operations. No significant difference was found between these two groups regarding the total resection rate and stratified radical resection rate according to different pathological types and cancer locations. The mean and median survival time of patients after radical operation in LSPH group were 13.9 ± 1.3 months and 14.8 months, respectively, while those in non-LSPH group were 22.6 ± 1.4 months and 18.4 months, respectively(P<0.05). CONCLUSIONS: Radical operations for pancreatic cancer and secondary LSPH are safe and effective. Because high-grade malignancy and poor prognosis are closely associated, the decision for radical surgery should be made more meticulously for the patients with pancreatic cancer.
Assuntos
Hipertensão Portal/cirurgia , Neoplasias Pancreáticas/patologia , Neoplasias Pancreáticas/cirurgia , Feminino , Humanos , Hipertensão Portal/fisiopatologia , Laparotomia/métodos , Masculino , Pessoa de Meia-Idade , Prognóstico , Estudos RetrospectivosRESUMO
AIM: To observe the effect of the human cytomegalovirus(HCMV)-encoded chemokine receptor homolog US28 on the human transcription factor CREB related transcriptional activity. METHODS: The US28 gene was cloned from DNA of HCMV-infected fibroblast at 72 h post infection. The amplified gene fragment was subsequently cloned into pcDNA3.1 eukaryotic expression vector. The recombinant plasmid was selected and identified by sequence analysis. US28-pcDNA3.1 was added to the Dual-Luciferase Reporter Assay System. The immunoreactive bands of phospho-CREB(p-CREB)and luminescence values were observed. RESULTS: The constructed recombinant vector was verified by PCR analysis and DNA sequencing. US28 enhanced the transcriptional efficiency of CRE driving gene via p-CREB. CONCLUSION: HCMV could enhance the transcriptional activity of CRE driving gene via p-CREB. CREB might be involved in the very early reprogramming of the host cell during virus activation.
Assuntos
Proteína de Ligação ao Elemento de Resposta ao AMP Cíclico/metabolismo , Citomegalovirus/metabolismo , Receptores de Quimiocinas/metabolismo , Proteínas Virais/metabolismo , Western Blotting , Linhagem Celular , Proteína de Ligação ao Elemento de Resposta ao AMP Cíclico/genética , Citomegalovirus/genética , Humanos , Luciferases/genética , Luciferases/metabolismo , Fosforilação , Reação em Cadeia da Polimerase , Receptores de Quimiocinas/genética , Proteínas Recombinantes de Fusão/genética , Proteínas Recombinantes de Fusão/metabolismo , Transcrição Gênica , Transfecção , Proteínas Virais/genéticaRESUMO
Protein microarray has progressed rapidly in the past few years, but it is still hard to popularize it in many developing countries or small hospitals owing to the technical expertise required in practice. We developed a cheap and easy-to-use protein microarray based on dot immunogold filtration assay for parallel analysis of ToRCH-related antibodies including Toxoplasma gondii, rubella virus, cytomegalovirus and herpes simplex virus type 1 and 2 in sera of pregnant women. It does not require any expensive instruments and the assay results can be clearly recognized by the naked eye. We analyzed 186 random sera of outpatients at the gynecological department with our microarray and commercial ELISA kit, and the results showed there was no significant difference between the two detection methods. Validated by clinical application, the microarray is easy to use and has a unique advantage in cost and time. It is more suitable for mass prenatal screening or epidemiological screening than the ELISA format.