Lentivirus mediated silencing of Ubiquitin Specific Peptidase 39 inhibits cell proliferation of human hepatocellular carcinoma cells in vitro

BACKGROUND: Ubiquitin Specific Peptidase 39 (USP39) is a 65 kDa SR-related protein involved in RNA splicing. Previous studies showed that USP39 is related with tumorigenesis of human breast cancer cells. RESULTS: In the present study, we investigated the functions of USP39 in human hepatocellular carcinoma (HCC) cell line SMMC-7721. We knocked down the expression of USP39 through lentivirus mediated RNA interference. The results of qRT-PCR and western blotting assay showed that both the mRNA and protein levels were suppressed efficiently after USP39 specific shRNA was delivered into SMMC-7721 cells. Cell growth was significantly inhibited as determined by MTT assay. Crystal violet staining indicated that colony numbers and sizes were both reduced after knock-down of USP39. Furthermore, suppression of USP39 arrested cell cycle progression at G2/M phase in SMMC-7721cells. In addition, Annexin V showed that downregulation of USP39 significantly increased the population of apoptotic cells. CONCLUSIONS: All our results suggest that USP39 is important for HCC cell proliferation and is a potential target for molecular therapy of HCC.

Computerized Tomography Contrast Induced Nephropathy (CIN) among adult inpatients / Nefropatia induzida por contraste (NIC) em pacientes adultos internados submetidos à tomografia computadorizada por contraste

Introduction: Contrast induced nephropathy (CIN) is one of the complications of the use of intravascular contrast agents, being defined as a reduction of the glomerular filtration rate caused by the iodinated contrast. Most CIN data derive from the cardiovascular literature, which identified as the most consistent risk factors pre-existing chronic renal insufficiency and diabetes mellitus. However, these studies limit their conclusions to a more specific patient population. Computerized tomography as a cause of CIN has been studied less often. Objective: To report on the incidence of computerized tomography contrast induced nephropathy (CIN) in an inpatient population of a tertiary general hospital, identifying potentially avoidable risk factors. Methods: We performed a prospective cohort study with inpatients admitted at a tertiary hospital requiring contrast-induced CT. The primary outcome was the development of CIN, measure by the alteration of serum creatinine or glomerular filtration rate in 48 or 72 hours. Through clinical interview, we verified possible risk factors and preventive measures instituted by the medical team and their association with development of CIN. Results: Of a total of 410 patients, 35 (8.5%) developed CIN. There was a positive correlation between CIN and the presence of diabetes mellitus (OR = 2.15; 95%CI 1.35-4.06; p = 0.02), heart failure (OR = 2.23; 95%CI 1.18-8.8; p = 0.022), and renal failure (OR = 3.36; 95%CI 1.57- 7.17; p = 0.002) Conclusion: Incidence of CIN varies according to the population. Diabetes mellitus, heart failure and renal failure were independent risk factors for the development of CT-associated CIN. Further studies are needed to better understand and treat CT-associated CIN. .

Introdução: Nefropatia induzida por contraste (NIC) é consequência do uso de meios de contraste intravenoso, sendo definida como uma redução da taxa de filtração glomerular. A maioria dos dados de NIC são da literatura cardiovascular, que identificou como fatores de risco insuficiência renal crônica e diabetes. Entretanto, esses estudos limitam suas conclusões a uma população especifica de pacientes. Tomografia Computadorizada contrastada como causa de NIC foi menos estudada. Objetivo: Reportar incidência de NIC numa população de pacientes internados em hospital terciário submetidos à tomografia computadorizada com contraste, identificando possíveis fatores de risco evitáveis. Métodos: Realizamos um estudo de coorte prospectivo com pacientes internados em hospital terciário e que necessitaram de tomografia computadorizada com contraste. O desfecho primário foi desenvolvimento de NIC, verificado por meio da variação da creatinina sérica ou taxa de filtração glomerular em 48 ou 72 horas. Em entrevista clínica, verificamos possíveis fatores de risco, assim como medidas preventivas instituídas pela equipe médica e suas possíveis associações com desenvolvimento de NIC. Resultados: Do total de 410 pacientes, 35 (8,5%) desenvolveram NIC. Houve correlação positiva entre desenvolvimento de NIC e a presença de diabetes mellitus (OR = 2,15; 95%CI 1,35-4,06; p = 0,02), insuficiência cardíaca (OR = 2,23; 95%CI 1,18-8,8; p = 0,022), e insuficiência renal (OR = 3,36; 95%CI 1,57-7,17; p = 0,002). Conclusão: A incidência de NIC varia de acordo com a população. Diabetes, insuficiência cardíaca e insuficiência renal foram fatores de risco independentes para o desenvolvimento de NIC. Mais estudos são ...

Role of complementary therapies in the understanding of primary healthcare professionals: a systematic review / Papel de prácticas complementarias en la comprensión de los profesionales de atención primaria: una revisión sistemática / Papel das práticas complementares na compreensão dos profissionais da Atenção Básica: uma revisão sistemática

Objective To identify the understanding of the healthcare professionals in relation to the role of complementary therapies in primary health care. Method Systematic review by way of the following information sources: PubMed, CINAHL, PeriEnf, AMED, EMBASE, Web of Science, Psicoinfo and Psicodoc, using the keyword Primary Health Care alone, and associated with the following keywords: Medicinal Plants, Herbal Medicine, Homeopathy, Traditional Chinese Medicine, Acupuncture, Anthroposophical Medicine. Results Twenty-two studies from 1986 to 2011 were included. We identified three styles of practice: conventional medicine, complementary therapies and integrative medicine. Positioning professional practices within these three styles may facilitate discussion of concepts of health care, enhancing the health care provided as a result. Conclusion The work process in primary care presents difficulties for conducting integrative and holistic health care, but this practice has been introduced over time by professionals who integrate conventional medicine and complementary therapies, concerned with the care and well-being of patients.
 .

Objetivo Identificar la comprensión de los profesionales sobre el papel de las prácticas complementarias en la atención primaria. Método Revisión sistemática. Fuentes de datos: PubMed, CINAHL, PeriEnf, AMED, EMBASE, Web of Science, Psicodoc y PysicoInfo. Descriptor Atención Primaria se asoció solo a los siguientes descriptores: plantas medicinales, fitoterapia, homeopatía, acupuntura, medicina tradicional china, medicina antroposófica. Resultados Se incluyeron 22 pesquisas entre 1986-2011. Tres estilos de práctica se identificaron: medicina convencional, medicina integrativa y Terapias Complementarias. Identificar la práctica profesional dentro de estos tres estilos puede facilitar la discusión de los conceptos de salud y la atención, mejorar la atención. Conclusión El proceso de trabajo en atención primaria presenta dificultades para realización de integración y atención integral, pero esta práctica se ha introducido con profesionales que integran medicina convencional y complementaria, ocupados con la atención y bienestar del paciente.


 .

Objetivo Identificar a compreensão dos profissionais de saúde quanto ao papel das práticas complementares na Atenção Básica. Método Revisão sistemática cujas fontes de informação foram: PubMed, CINAHL, PeriEnf, AMED, EMBASE, Web of Science, PysicoInfo e PsicoDoc, utilizando o descritor Atenção Básica associado, isoladamente, aos seguintes descritores: Plantas Medicinais, Fitoterapia, Homeopatia, Medicina Tradicional Chinesa, Acupuntura, Medicina Antroposófica. Resultados Incluíram-se 22 estudos entre 1986-2011. Identificaram-se três estilos de prática: medicina convencional, práticas integrativas e medicina integrativa. Posicionar a prática profissional dentro desses três estilos pode facilitar a discussão de concepções de saúde e cuidado, ampliando o cuidado. Conclusão O processo de trabalho na Atenção Básica apresenta dificuldades para a realização de cuidado integrativo e holístico, mas essa prática vem sendo introduzida com profissionais que integram medicina convencional e práticas complementares, preocupados com o cuidado e o bem-estar do paciente.

 .

Newer treatments for advanced hepatocellular carcinoma

Hepatocellular carcinoma (HCC) is one of the most common cancers worldwide. The only curative treatment modalities for HCC are surgery, percutaneous ablation, and liver transplantation. Unfortunately, the majority of patients have unresectable disease at diagnosis. Therefore, effective treatment options are needed for patients with advanced HCC. The current standard treatment for patients with advanced HCC, according to the Barcelona Clinic Liver Cancer staging system, is the multikinase inhibitor sorafenib. Other alternative therapies are required, due to the limited treatment response to, and tolerance of, this molecular target agent. Clinical trials of hepatic artery infusion chemotherapy, radioembolization, and multimodal treatments have shown favorable results in advanced HCC patients. This article introduces new treatment modalities for advanced HCC and discusses future therapeutic possibilities.

A novel prognostic factor for hepatocellular carcinoma: protein disulfide isomerase

BACKGROUND/AIMS: Protein disulfide isomerase (PDI) has been implicated in the survival and progression of some cancer cells, by compensating for endoplasmic reticulum stress by upregulating the protein-folding capacity. However, its prognostic role in patients with hepatocellular carcinoma (HCC) has not been investigated. METHODS: We collected HCC tissues from 83 HCC patients who underwent surgical resection for an immunohistochemical study of PDI. Overall survival (OS) was measured from the date of surgical resection until the date of death from any cause. Radiological progression was evaluated using the modified Response Evaluation Criteria in Solid Tumors in an independent radiological assessment. RESULTS: PDI expression was found to be increased in human HCC compared to adjacent nontumor tissues. Increased immunopositivity for PDI was associated with a high Edmondson-Steiner grade (p = 0.028). Univariate analysis of patients who had undergone surgical resection for HCC showed that tumor PDI upregulation is a significant risk factor for poor OS (p = 0.016; hazard ratio [HR], 1.980) and time to progression (TTP; p = 0.007; HR, 1.971). Multivariate analyses revealed that high PDI expression was an independent predictor of a shorter TTP (p = 0.015; HR, 1.865) and poor OS (p = 0.012; HR, 2.069). CONCLUSIONS: Upregulated PDI expression is associated with aggressive clinicopathological features of HCC; thus, PDI might serve as an independent prognostic factor and a potential therapeutic target for HCC patients.

Pro-oncogenic potential of NM23-H2 in hepatocellular carcinoma

NM23 is a family of structurally and functionally conserved proteins known as nucleoside diphosphate kinases (NDPK). There is abundant mRNA expression of NM23-H1, NM23-H2, or a read through transcript (NM23-LV) in the primary sites of hepatocellular carcinoma (HCC). Although the NM23-H1 protein is implicated as a metastasis suppressor, the role of NM23-H2 appears to be less understood. Thus, the aim of this study was to examine whether NM23-H2 is associated with hepatocarcinogenesis. The level of NM23-H2 expression in tumor tissues and the surrounding matrix appeared to be independent of etiology and tumor differentiation. Its subcellular localization was confined to mainly the cytoplasm and to a lesser extent in the nucleus. Ectopic expression of NM23-H2 in NIH3T3 fibroblasts and HLK3 hepatocytes showed a transformed morphology, enhanced focus formation, and allowed anchorage-independent growth. Finally, NIH3T3 fibroblasts and HLK3 hepatocytes stably expressing NM23-H2 produced tumors in athymic mice and showed c-Myc over-expression. In addition, NF-kappaB and cyclin D1 expression were also increased by NM23-H2. Lentiviral delivery of NM23-H2 shRNA inhibited tumor growth of xenotransplanted tumors produced from HLK3 cells stably expressing NM23-H2. Collectively, these results indicate that NM23-H2 may be pro-oncogenic in hepatocarcinogenesis.

IQGAP1 is overexpressed in hepatocellular carcinoma and promotes cell proliferation by Akt activation

The scaffold protein IQGAP1 shows elevated levels in several cancer types, but its expression in hepatocellular carcinoma is unknown. We found that 58% of human hepatocellular carcinoma tissue samples had increased IQGAP1 expression compared to adjacent normal tissue. Overexpressing IQGAP1 raised the in vivo tumorigenicity of hepatocellular carcinoma cells, and forced overexpression of IQGAP1 in vitro stimulated cell proliferation. Cell growth was reduced by knockdown or mutation of IQGAP1, or by treatment of cells with a phosphotidylinositol 3-kinase inhibitor. To determine the mechanism by which IQGAP1 overexpression affected hepatocellular carcinoma cells, we confirmed its interaction in these cells with mammalian target of rapamycin (mTOR), a serine/threonine kinase that integrates signals about nutrient and energy status with downstream effectors that influence cell division. In addition, we discovered a new interaction involving IQGAP1, mTOR and Akt, which is a downstream target of mTOR. Akt phosphorylation on Ser-473, which is catalyzed by mTOR and required for Akt activation, increased with increasing amounts of IQGAP1, and decreased with IQGAP1 mutation. We hypothesize that IQGAP1 is a scaffold that facilitates mTOR and Akt interaction.

Lack of influence of cyclooxygenese-2 expression in hepatocellular carcinomas on patient survival.

Recent studies suggest that cyclooxygenese-2 (COX-2) enzyme activation may play a role in hepatocarcinogenesis. However, the clinical significance of COX-2 expression in hepatocellular carcinoma (HCC) remains obscure. This study evaluated COX-2 expression in hepatitis B and hepatitis C virus related HCC and in HCC patients with an unknown etiology. Liver tissue samples of 31 patients with HCC (27 men and 4 women; age range, 48-75 years) were analyzed. COX-2 expression was evaluated by immunohistochemically in the tumor tissues. Patient data including age, sex, Child score, stage, grade of the tumor and survival were analyzed. Of these patients 19 were positive for hepatitis B virus (HBV), 6 were positive for hepatitis C virus (HCV) and 6 patients were negative for all viral markers and other etiologic factors. COX-2 staining were evaluated in 2 groups (group 1: COX-2 expression less than 25% (grades 1-2 COX-2 expression), and group 2: Cox-2 expression 25% or more (grades 3-5 COX-2 expression). COX-2 expression was shown in all HCC samples with positive or negative viral markers. No difference was found between degree of COX-2 expression and the etiology of HCC. COX-2 expression was not correlated with number of lesion or stage of the disease or grade of the tumor. COX-2 expression was not related with Child score of the patients. Median survival of all patients was 32 months. Median survival of patients did not differ according to patient's viral marker status. No difference was observed in median survival of patients in group 1 and 2. As a result, COX-2 system seem to be shared part in hepatocarcinogenesis regardless factors that initiate the disease. Although COX-2 expression appears to be independent of disease's characteristics', treatments that target this system appear to be feasible in the management of HCC.

Detection of hTERT mRNA in gastrointestinal tract cancer specimens.

Human telomerase consisting of telomerase RNA template (hTR) and telomerase reverse transcriptase (hTERT) provides a mechanism for synthesis of telomere repeats that prolongs life span of cells. Telomerase activity is present in germ-line and malignant tumor cells but not in most normal human somatic cells. This study determined hTERT mRNA level in tissue samples from patients with gastrointestinal tract (GI) cancers. Tissue samples were obtained from 22 GI cancer patients, 3 gastrointestinal stomal tumors (GIST) and 25 corresponding non-cancerous tissues. hTERT expression was determined by real-time reverse transcriptase-polymerase chain reaction (RT-PCR) using Taqman probe, hTERT mRNA was detected in 12 of 22 cancerous tissue samples. Six of 8 tissue samples obtained from patients with hepatocellular carcinoma and cholangiocarcinoma were positive for hTERT. However, hTERT mRNA was not detected in GIST and non-cancerous tissues. These results suggest that hTERT may be an effective target for cancer therapies to treat many type of GI cancers including cholangiocarcinoma and hepatocellular carcinoma.

Down-regulation of survivin in growth inhibition of hepatoma cells induced by a selective cyclooxygenase-2 inhibitor / 대한간학회지

BACKGROUND/AIMS: Cyclooxygenase-2 (COX-2) inhibitors reportedly inhibit the growth of hepatocellular carcinoma (HCC) via caspase-dependent or caspase-independent apoptosis, which is due to COX-2 being associated with hepatocarcinogenesis. Survivin is highly expressed in most human cancers, but the mechanism regulating survivin expression remains unclear. We investigated the regulatory expression of survivin in selective-COX-2-inhibitor-induced growth inhibition of hepatoma cells. METHODS: After treatment with NS-398 (a selective COX-2 inhibitor) at various concentrations (10, 50, 100, 150, and 200 micrometer), the growth inhibition of Hep3B hepatoma cells was assessed by an MTT cell-viability assay, DNA fragmentation gel analysis, and flow cytometry. The expression of survivin transcript was analyzed by reverse-transcription polymerase chain reactions. RESULTS: NS-398 inhibited the growth of hepatoma cells by an amount dependent on the concentration and the time since treatment. Apoptotic DNA ladder and flow-cytometry shifting to the sub-G1 phase were revealed in NS-398-induced growth inhibition of hepatoma cells. NS-398 suppressed the expression of the survivin gene in a concentration- and time-dependent manner. CONCLUSIONS: Survivin was down-regulated in the growth inhibition of hepatoma cells induced by a selective COX-2 inhibitor, NS-398, in a concentration- and time-dependent manner. These results suggest the therapeutic inhibition of COX-2 via suppression of survivin in HCC.

Anti-tumor mechanisms and regulation of survivin by selective cyclooxygenase-2 inhibitor / 대한간학회지

No abstract available.

Correlative analysis of histological profile of the adjoining liver parenchyma with liver enzyme levels in hepatocellular carcinoma and their comparison with chronic liver disease in autopsy cases.

The exact mechanism and aetiological factor for hepatocarcinogenesis is not yet well defined. Besides genomic integration of hepatitis B viral particles, persistent chronic inflammation is postulated to be important initiating factor in viral related hepatocellular carcinoma (HCC). The objectives of the present study were--to correlate histological profiles of chronic liver disease in the adjoining non-tumor liver tissue in HCC with liver enzymes, to compare with those of non-carcinomatous chronic liver disease cases using the liver tissue and data collected at autopsy, and to correlate with hepatitis B and C positive status. Post mortem liver and data available at autopsy were used for the study. Changes of chronic liver disease was graded and staged according to Peter Scheur's (1991). In HCC, the non-malignant liver tissue was used for the study. Hepatitis B surface and core antibodies were demonstration by immunohistochemistry. HCV was documented by RT-PCR using the tissue extract of paraffin embedded liver tissue. HCC group had higher inflammatory grading and transaminases levels than non-HCC group. HBcAg alone and dual HBcAg and HCV positive cases were more in HCC group. Incidence of biliary epithelial cell injury was higher in HCV positive subgroup. Conclusion: higher incidence of inflammatory grading and enzyme level in alone HBcAg and dual HBcAg and HCV positivity in HCC group would suggest significant role of ongoing persistent chronic inflammation and actively replicating HBV and HCV infections in carcinogenesis.

Regulation of cell growth by fatty acid-CoA ligase 4 in human hepatocellular carcinoma cells

Fatty acid-CoA ligase 4 (FACL4) is a central enzyme controlling the unesterified free arachidonic acid (AA) level in cells and the free AA is known to induce apoptosis. We have recently reported that expression of FACL4 is upregulated in about 40% of human hepatocellular carcinoma (HCC) and 50% of HCC cell lines, suggesting that FACL4 may be involved in liver carcinogenesis. In this study, we investigated whether HCC cell growth is regulated by FACL4. Immunoblot analysis showed that SNU 398 cells express very low or no detectable level of FACL4. We, therefore, transfected the SNU 398 cells with FACL4 expression vector, and clones expressing FACL4 were pooled and analyzed. We found that forced expression of FACL4 in SNU 398 promotes the growth of cells. In addition, we observed that triacsin C, a FACL4 inhibitor, inhibits the growth of Hep 3B cell line which expresses high level of endogenous FACL4. We also found that the triacsin C-mediated growth inhibition in Hep 3B cells results from the induction of apoptosis with evidence of Bcl-2 reduction. Altogether, our data show that FACL4 affects HCC cell growth and suggest that modulation of FACL4 expression/activity is an approach for treatment of HCC.

Telomerase is strongly activated in hepatocellular carcinoma but not in chronic hepatitis and cirrhosis

Telomerase is highly activated in human immortal cell lines and tumor tissues, whereas it is not activated in primary cell strains and many tumor-adjacent tissues. It is suggested that telomerase activation is one of the critical steps in malignant transformation. In the present study, the telomerase activity was investigated in hepatocellular carcinoma tissues and non-tumor liver tissues from Korean patients with chronic hepatitis and cirrhosis. Eighty two liver tissues (24 chronic hepatitis specimens, 34 cirrhosis specimens, and 24 hepatocellular carcinomas) were obtained from 23 chronic viral hepatitis patients, 19 cirrhosis patients (including 7 liver transplants), and 24 patients with hepatocellular carcinoma, of which the surrounding non-tumor liver tissues were available in 16 patients (1 chronic hepatitis and 15 cirrhosis). As negative controls, 3 normal liver tissues were included. Protein from liver specimens was purified by a detergent lysis method as described elsewhere, and telomerase activity was measured in 2 diluents of each sample (1:1 and 1:100) by a telomeric repeat amplification protocol (TRAP). Telomerase was strongly activated in 79% (19/24) of the hepatocellular carcinomas, while weakly in 8% (2/24) of the chronic hepatitis tissues and in 24% (8/34) of the cirrhosis tissues. All of 3 normal control livers showed no telomerase activation. No relationship could be observed between the enhancement of telomerase activity and tumor nature. None of the chronic heaptitis or cirrhosis patients with mild telomerase activation in the liver have developed hepatocellular carcinoma for at least 2 years of follow-up period. These results suggest that the strong enhancement of telomerase activity may be a critical part of hepatocarcinogenesis, although the exact mechanism of such high activation in hepatocellular carcinoma is not clear. In addition, further study will be necessary to clarify the reason why no telomerase activity detectable by a conventional TRAP can be seen in some hepatocellular carcinoma.

Deficiencia de alfa-1 antitripsina: presentación de un estudio familiar

Se informa el caso de un paciente de 65 años de edad quien presentaba un hepatocarcinoma y cirrosis asociados a deficiencia de alfa-1antitripsina (A1-AT). Cuatro de sus seis hermanos, dos hombres y dos mujeres, tenían valor de esta glicoproteína inferiores a los normales, sin manifestación aparente de compromiso clínico hepático ni pulmonar; ninguno tenía antecedentes de hepatitis o colestasis neonatal. La descripción de este primer caso en nuestro medio se propone estimular el interés por esta entidad como causa de hepatopatía crónica, carcinoma hepático primario y enfisema pulmonar, sin factores de riesgo diferentes asociados