RESUMO
Hepatocellular carcinoma (HCC) is one of the most common malignant tumors in the human digestive system, and has been recognized as a serious threat to public health worldwide. This study explored the role of chondroitin polymerizing factor (CHPF) in the development and metastasis of HCC. Immunohistochemistry analysis was performed to detect CHPF expression in HCC tissues and para-carcinoma tissues. qRT-PCR and Western blot analysis were used to determine the mRNA and protein expression of CHPF. MTT assays, colony formation assays, and flow cytometry were used to evaluate the cell proliferation, colony formation, and cell apoptosis, respectively. Wound-healing and Transwell assays were performed to evaluate cell migration. The results show that CHPF was not only up-regulated in HCC tissues compared with para-carcinoma tissues, but was also related with more advanced stages of HCC. Further studies revealed that CHPF knockdown significantly inhibited cell proliferation and colony formation, and induce cell apoptosis of HCC cells. Moreover, suppressing the expression of CHPF reduced the migration and invasiveness of HCC cells. In conclusion, we demonstrated that CHPF plays important roles in the development and progression of HCC, and high expression levels of HCC may be related with poorer prognosis. The results from this study may provide a potential therapeutic target for HCC treatment.
Assuntos
Carcinoma Hepatocelular/patologia , Regulação Neoplásica da Expressão Gênica , Neoplasias Hepáticas/patologia , N-Acetilgalactosaminiltransferases/metabolismo , Apoptose , Carcinoma Hepatocelular/metabolismo , Linhagem Celular Tumoral , Movimento Celular , Proliferação de Células , Citometria de Fluxo , Perfilação da Expressão Gênica , Humanos , Imuno-Histoquímica , Neoplasias Hepáticas/metabolismo , Metástase Neoplásica , Prognóstico , RNA Mensageiro/metabolismo , Regulação para Cima , CicatrizaçãoRESUMO
Kruppel-like factor 8 (KLF8) is a transcription factor which has been identified to play a critical role in oncogenic transformation, epithelial-mesenchymal transition and invasion. Higher expression level of KLF8 has been observed in ovarian, renal and breast cancer cells. This study focused on investigating the knockdown effects of KLF8 through lentivirus mediated targeted disruption of KLF8 in gastric cancer cell lines. The expression level of KLF8 is much higher in gastric cancer cells than that in normal cell via Western blot analysis. The decreased expression level of KLF8 after repression was confirmed by real-time PCR and Western blot in SGC-7901, a gastric cancer cell line. The effects of KLF8 deletion on cell proliferation and cell cycle were analyzed by MTT assay and flow cytometry, respectively. Finally, the effects of KLF8 deletion on cell apoptosis and invasion of gastric cancer cells were analyzed by Annexin staining and transwell assay, respectively. It was observed that knockdown of KLF8 reduced the cellular proliferation of SGC-7901 gastric cancer cells, a phenotype at least partially due to cell cycle arrest at G1 phase and increased apoptosis. Furthermore, the inhibition of KLF8 reduces the invasion rates of the cancer cells. Therefore, KLF8 is necessary for cell survival and invasion in gastric cancer cells. The absence of KLF8 may lead to cancer cell death. These results demonstrated that the lentivirus mediated targeted disruption of KLF8 would be an promising therapeutic method for treatment of gastric cancer.
Assuntos
Proliferação de Células , Lentivirus/genética , Interferência de RNA , Proteínas Repressoras/genética , Adenocarcinoma , Apoptose , Pontos de Checagem do Ciclo Celular , Linhagem Celular Tumoral , Movimento Celular , Técnicas de Silenciamento de Genes , Vetores Genéticos , Humanos , Fatores de Transcrição Kruppel-Like , Proteínas Repressoras/metabolismo , Neoplasias GástricasRESUMO
BACKGROUND: This study aims to explore the relationship between spleen arterial blood flow (SBF) with platelet count, spleen index (SPI) and the serum nitric oxide (NO) level of patients with liver cirrhosis and to investigate the role of SBF in the development of hypersplenism. METHODOLOGY: Platelet count, SPI, SBF and serum NO levels were evaluated in 100 patients with liver cirrhosis caused by hepatitis B with hypersplenism (cirrhosis group) and 30 healthy persons without hypersplenism (control group). RESULTS: Platelet count in cirrhosis group and control group was 57.0 +/- 25.6 x 109/L and 205.8 +/- 47.4 x 109/L (p = 0.000), SBF was 535.7 +/- 263.7 milmin and 172.2 +/- 66.9 ml/min (p = 0.000), and serum NO level was 98.51 +/- 23.06 micromol/L and 48.43 +/- 19.47 micromol/L (p = 0.000). Linear correlations were made between SBF and platelet count in cirrhosis group (r = -0.573, p = 0.000), SBF and SPI (r = 0.607, p = 0.01), SBF and serum NO level (r = 0.754, p = 0.000). Moreover, serum NO level increased as liver disease aggravated (82.50 +/- 15.04 pmol/L in Child grade A, 94.61 +/- 21.00 micromol/L in grade B and 116.83 +/- 18.03 micromol/L in grade C; grade A versus grade C, p = 0.003). CONCLUSION: The elevation of SBF may play an important role in the development of hypersplenism and disorders in vasoactive factors such as the serum NO caused by liver cirrhosis may play an important role in the elevation of SBF.
Assuntos
Hiperesplenismo/fisiopatologia , Artéria Esplênica/fisiologia , Adulto , Idoso , Velocidade do Fluxo Sanguíneo/fisiologia , Estudos de Casos e Controles , Feminino , Hepatite B/complicações , Humanos , Hiperesplenismo/sangue , Hiperesplenismo/diagnóstico por imagem , Hiperesplenismo/etiologia , Cirrose Hepática/virologia , Masculino , Pessoa de Meia-Idade , Óxido Nítrico/sangue , Contagem de Plaquetas , Índice de Gravidade de Doença , Ultrassonografia DopplerRESUMO
Circular RNAs (circRNAs) are critical regulators in tumor initiation and development and participate in the pathological process of hepatocellular carcinoma (HCC). However, the specific role and mechanism of circRNA, hsa_circ_102559, in HCC remains elusive. First, analysis of HCC-related circRNA expression profile GSE97332 and HCC patients showed a significant upregulation of hsa_circ_102559 in HCC tissues. Upregulation of hsa_circ_102559 in HCC cells was associated with the metastatic properties. Second, hsa_circ_102559 significantly promoted HCC metastasis, while knockdown of hsa_circ_102559 reversed the promotive effects on HCC progression. Functionally, hsa_circ_102559 could target and colocalize with miR-130a-5p in the cytoplasm of HCC cells. Annexin A2 (ANXA2) was identified as a target gene of miR-130a-5p, and overexpression of ANXA2 counteracted with the suppressive effects of hsa_circ_102559 silence on HCC metastasis. Lastly, xenograft experiment was established and results indicated that knockdown of hsa_circ_102559 inhibited HCC growth and metastasis through the downregulation of ANXA2. In conclusion, hsa_circ_102559 inhibited HCC progression via sponging miR-130a-5p to reduce ANXA2 expression, suggesting that hsa_circ_102559 might be a potential biomarker or therapeutic target for HCC.
Assuntos
Anexina A2/metabolismo , Carcinoma Hepatocelular/metabolismo , Carcinoma Hepatocelular/patologia , Neoplasias Hepáticas/metabolismo , Neoplasias Hepáticas/patologia , MicroRNAs/metabolismo , Anexina A2/genética , Humanos , MicroRNAs/genéticaRESUMO
Ribosomal protein s15a (RPS15A) plays a promotive role in the mRNA/ribosome interactions during early translation. Our previous study has found that inhibiting RPS15A expression can decrease proliferation and induce cell cycle arrest in hepatocellular carcinoma (HCC) cell lines. However, the mechanism underlying the involvement of RPS15A in HCC pathogenesis and the clinical significance of RPS15A expression remain unclear. In this study, an evaluation of RPS15A expression in 110 surgically resected HCCs and matched tumor-adjacent normal tissues revealed an overexpression of RPS15A in HCC, which was correlated with worse survival. In addition, tumor tissue with higher RPS15A expression demonstrated a higher microvascular density (MVD). Subsequently, two HCC cell lines, Huh7 (low-level constitutive RPS15A expression) and HepG2 (high RPS15A expression) were used to further evaluate the role of RPS15A in angiogenesis. The co-culture experiment of HCC cells with endothelial cells revealed that the induced overexpression of RPS15A in Huh7 cells increased the angiogenic potential of HUVEC in a paracrine fashion; conversely, knockdown of RPS15A in HepG2 cells showed an opposite effect. Further analysis indicated that RPS15A modulated FGF signaling by enhancing Wnt/beta-catenin-mediated FGF18 expression in HCC cells. FGF18, in turn, through binding to its FGFR3 receptor on endothelial cells, can activate the AKT and ERK pathway and promotes angiogenesis in a tumor microenvironment. Our in vivo experiment further confirmed that inhibition of RPS15A expression in HCC xenografts dramatically hindered tumor growth and inhibited tumor angiogenesis. Together, our findings suggest that RPS15A promotes angiogenesis in HCCs by enhancing Wnt/beta-catenin induced FGF18 expression. The RPS15A/FGF18 pathway may be a rational target for anti-angiogenic therapy of HCC.
Assuntos
Carcinoma Hepatocelular/irrigação sanguínea , Fatores de Crescimento de Fibroblastos/metabolismo , Neoplasias Hepáticas/irrigação sanguínea , Neovascularização Patológica/patologia , Proteínas Ribossômicas/metabolismo , Proteína Wnt1/metabolismo , beta Catenina/metabolismo , Adulto , Idoso , Animais , Apoptose , Biomarcadores Tumorais/genética , Biomarcadores Tumorais/metabolismo , Carcinoma Hepatocelular/metabolismo , Carcinoma Hepatocelular/patologia , Estudos de Casos e Controles , Proliferação de Células , Feminino , Fatores de Crescimento de Fibroblastos/genética , Seguimentos , Regulação Neoplásica da Expressão Gênica , Humanos , Neoplasias Hepáticas/metabolismo , Neoplasias Hepáticas/secundário , Metástase Linfática , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Nus , Pessoa de Meia-Idade , Invasividade Neoplásica , Neovascularização Patológica/genética , Neovascularização Patológica/metabolismo , Prognóstico , Proteínas Ribossômicas/genética , Taxa de Sobrevida , Células Tumorais Cultivadas , Proteína Wnt1/genética , Ensaios Antitumorais Modelo de Xenoenxerto , beta Catenina/genéticaRESUMO
The antidiabetic drug metformin has been shown to possess antitumor functions in many types of cancers. Although studies have revealed its beneficial effects on the prognosis of hepatocellular carcinoma (HCC), the detailed molecular mechanism underlying this event remains largely unknown. In this work, we showed that miR-23a was significantly induced upon metformin treatment; inhibition of miR-23a abrogated the proapoptotic effect of metformin in HepG2 cells. We next established forkhead box protein A1 (FOXA1) as the functional target of miR-23a, and silencing FOXA1 mimicked the effect of metformin. Moreover, the phosphorylation of AMP-activated protein kinase (AMPK) and the expression of p53 were increased upon metformin treatment, and the inhibition of p53 abrogated the induction of miR-23a by metformin, suggesting that AMPK/p53 signaling axis is responsible for the induction of miR-23a by metformin. In summary, we unraveled a novel AMPK/p53/miR-23a/FOXA1 axis in the regulation of apoptosis in HCC, and the application of metformin could, therefore, be effective in the treatment of HCC.
RESUMO
BACKGROUND: Endothelial progenitor cells (EPCs) can migrate to the tumor tissue and enhance the angiogenesis of hepatocellular carcinoma (HCC); thus, they are associated with a poor prognosis. However, the specific molecular mechanism underlying the homing of EPCs to the HCC neovasculature remains unrevealed. METHODS: Co-culture experiments of endothelial progenitor cells with HCC cells with modulation of EphA1 were performed in vitro. Using EPCs as angiogenic promoters by injecting them into HCC xenograft-bearing nude mice via their tail veins to test homing ability of EPCs changed according to different EphA1 level in HCC xenograft. RESULTS: In this study, we found that the up-regulation of EphA1 expression in HCC cells could affect not only the chemotaxis of EPCs to tumor cells and endothelial cells (ECs) but also the tube formation ability of EPCs in a paracrine fashion. Further, we revealed that the increased expression of EphA1 in HCC cells led to an increased SDF-1 concentration in the tumor microenvironment, which in turn activated the SDF-1/CXCR4 axis and enhanced the recruitment of EPCs to HCC. In addition, the EphA1-activated SDF-1 expression and secretion was partially mediated by the PI3K and mTOR pathways. In vivo experiments demonstrated that blocking EphA1/SDF-1/CXCR4 signaling significantly inhibited the growth of HCC xenografts. Using immunohistochemistry and immunofluorescence assays, we verified that the inhibition of tumor angiogenesis was at least partially caused by the decreased number of EPCs homing to tumor tissue. CONCLUSIONS: Our findings indicate that targeting the EphA1/SDF-1 signaling pathway might be a therapeutic anti-angiogenesis approach for treating HCC.
Assuntos
Carcinoma Hepatocelular/patologia , Quimiocina CXCL12/metabolismo , Células Endoteliais/citologia , Neoplasias Hepáticas/patologia , Neovascularização Patológica/metabolismo , Receptor EphA1/metabolismo , Receptores CXCR4/metabolismo , Animais , Carcinoma Hepatocelular/genética , Carcinoma Hepatocelular/metabolismo , Linhagem Celular Tumoral , Movimento Celular , Quimiocina CXCL12/genética , Técnicas de Cocultura , Células Endoteliais/transplante , Células Endoteliais da Veia Umbilical Humana , Humanos , Neoplasias Hepáticas/genética , Neoplasias Hepáticas/metabolismo , Camundongos , Neovascularização Patológica/genética , Neovascularização Patológica/patologia , Receptores CXCR4/genética , Transdução de Sinais , Microambiente TumoralRESUMO
Glypican-3 (GPC3) is a widely used immunohistochemical marker for hepatocellular carcinoma (HCC); however, its prognostic value is unclear. Immunohistochemical evaluation of GPC3 expression was performed on 300 postoperative HCC tissue samples with paired adjacent non-tumor tissues on tissue microarray sections. The integral optic density, representing the expression level of GPC3 in each HCC sample, was calculated using Image-Pro Plus. The outcome-based cut-point optimization was performed using X-tile software. GPC3 was highly expressed in HCC tissues compared with adjacent non-tumor tissues. The expression level of GPC3 was significantly correlated with overall survival (OS) and time to recurrence (TTR). The lower the level of GPC3 expression in HCC tissue, the poorer the observed prognosis. Univariate and multivariate analyses showed that the expression level of GPC3 in HCC was an independent prognostic factor for both OS and TTR. In conclusion, GPC3 expression is an independent prognostic factor for postoperative HCC, and low expression levels of GPC3 in HCC may indicate poor outcome.
Assuntos
Biomarcadores Tumorais/análise , Carcinoma Hepatocelular/patologia , Glipicanas/análise , Interpretação de Imagem Assistida por Computador/métodos , Neoplasias Hepáticas/patologia , Adulto , Idoso , Carcinoma Hepatocelular/metabolismo , Feminino , Humanos , Imuno-Histoquímica , Estimativa de Kaplan-Meier , Neoplasias Hepáticas/mortalidade , Masculino , Pessoa de Meia-Idade , Análise Multivariada , Prognóstico , Modelos de Riscos Proporcionais , Software , Análise Serial de TecidosRESUMO
OBJECTIVE: To discuss the value of multiple-slice spiral CT diagnosis of atypical intraabdominal hernia. METHODS: The clinical and CT findings of 16 cases of atypical intraabdominal hernia confirmed surgically were retrospectively analysed. RESULTS: In all the 16 cases, the contents of hernia were small bowels and the mesentery. Nine cases were caused by the adhesion after abdominal operations or infection, 6 by the mesenteric foramen, and 1 by the gap of pelvic peritoneum. The main CT findings were as follows:(1)the obstructed small bowels gathered abnormally and showed cluster shape(9 cases); the walls of the bowels thickened with edema and showed "target" sign with exudate in the neighboring spaces(5 cases); other part of the abdominal cavity became empty for lack of small bowels(4 cases).(2) Abnormal arrangement of the branches of mesenteric vessel, which appeared gathered or pulled or rigid and displacement of the main branches to left or right(12 cases). Thickened mesenteric vessel (4 cases): torsion of mesentery with "whirlpool" sign (3 cases). (3) With the help of multiplanar reformation, 5 cases showed the evidence of hernia rings. In the ring area, there were gathered or pulled or rigid and radiating mesenteric vessel and the dilated or effusion bowels in cluster arrangement forming "parachute" and "bundle of balloons" sign. CONCLUSION: CT manifestations of atypical intraabdominal hernia has some specific characteristics, which is of important value for clinical diagnosis and treatment.