RESUMO
OBJECTIVE: Tumor necrosis factor-associated apoptosis-inducing ligand (TRAIL) is a potent anticancer agent, which could specifically target cancerous cells. Nutlin-3, a small-molecular inhibitor of murine double minute 2 (MDM2), shows oncogenic potential in a variety of human cancers. It has also been found to promote the TRAIL-induced apoptosis of cancer cells in esophageal squamous cancer, but its potential role and underlying mechanisms in the TRAIL-treated hepatocellular carcinoma (HCC) remains to be elucidated. METHODS: HSS cell line (Huh7) cells were used as an in vitro model of HCC. TRAIL (100 ng/ml) was used to induce cell apoptosis. Cell viability was measured by CCK-8 assay. Cell apoptosis was detected using TUNEL staining. Mitochondrial activity was evaluated by measuring caspase 3/7 and 9 levels. P53 expression at protein and mRNA level were measured by Western blotting and RT-qPCR, respectively. RESULTS: The combination of Nutlin-3 and TRAIL facilitated the apoptosis and increased the levels of mitochondrial cleaved-caspase3/7 and 9 in HCC cells compared with TRAIL treatment alone, both in a concentration-dependent way. Nutlin-3 also upregulated the expression of p53 and DR5, while knockdown of p53 significantly hindered the pro-apoptotic effect of Nutlin-3. Further studies revealed that Nutlin-3 downregulated the expression of survivin and bcl-2, both of which could be reversed by p53 knockdown. Moreover, survivin suppressant YM155 and bcl-2 inhibitor YM155 further enhanced the apoptosis of HCC cells in the presence of Nutlin-3 and TRAIL. CONCLUSION: These results suggested that Nutlin-3 facilitated TRAIL-induced apoptosis of HCC cells by activating the p53-survivin/bcl-2 pathway, which provided novel insights into the mechanism of Nutlin-3 and confirmed the potential of combination of Nutlin-3 and TRAIL as an adjuvant in HCC therapy.
Assuntos
Antineoplásicos , Carcinoma Hepatocelular , Neoplasias Hepáticas , Animais , Antineoplásicos/farmacologia , Apoptose , Carcinoma Hepatocelular/tratamento farmacológico , Carcinoma Hepatocelular/genética , Carcinoma Hepatocelular/patologia , Caspase 3/metabolismo , Linhagem Celular Tumoral , Humanos , Imidazóis , Ligantes , Neoplasias Hepáticas/patologia , Camundongos , Piperazinas , Proteínas Proto-Oncogênicas c-bcl-2/metabolismo , Proteínas Proto-Oncogênicas c-mdm2/metabolismo , RNA Mensageiro , Survivina/metabolismo , Ligante Indutor de Apoptose Relacionado a TNF/genética , Ligante Indutor de Apoptose Relacionado a TNF/farmacologia , Proteína Supressora de Tumor p53/metabolismoRESUMO
Patients with catheter-related bloodstream infection (CRBSI) have a poor prognosis. Proadrenomedullin (pro-ADM) has emerged as a valuable marker of sepsis. The potential role of pro-ADM in predicting the prognosis of CRBSI was evaluated. We enrolled 25 CRBSI patients and pro-ADM level was measured within 24âhours after each admission. Survival was assessed after 28 days. Among 25 patients with CRBSI, 14 patients survived. Pro-ADM in survivors was significantly lower than that in non-survivors (3.71â±â1.30 vs 5.58â±â1.18ânmol/L). The area under the curve (AUC) for pro-ADM was 0.87 (95% CI 0.68-0.97) with a cut-off value of 4.67ânmol/L, providing sensitivity of 85.7% and specificity of 81.8%. The AUCs for PCT, WBC, and CRP were 0.76 (95% CI 0.55-0.90), 0.72 (95% CI 0.50-0.88), and 0.69 (95% CI 0.48-0.86), respectively. Kaplan-Meier survival curves showed pro-ADM ≥ 4.67ânmol/L was associated with higher mortality (log-rank pâ=â0.001). Moreover, the pro-ADM level was significantly higher in patients with septic shock than those without shock (5.44â±â1.17 vs 3.54â±â1.18nmol/L). The mortality of patients with septic shock was higher than that of patients without shock (69.2% vs 16.7%, Pâ=â.008). In conclusion, pro-ADM could be used as a prognostic marker of CRBSI in critically ill patients.
