AIM:To assess effects of heme on messenger RNA(mRNA) and microRNA(miRNA) profiles of liver cells derived from humans.METHODS:We exposed human hepatoma cell line Huh-7 cells to excess iron protoporphyrin(heme)(10 μmol...AIM:To assess effects of heme on messenger RNA(mRNA) and microRNA(miRNA) profiles of liver cells derived from humans.METHODS:We exposed human hepatoma cell line Huh-7 cells to excess iron protoporphyrin(heme)(10 μmol/L) or induced heme deficiency by addition of 4,6-dioxoheptanoic acid(500 μmol/L),a potent inhibitor of aminolevulinic acid dehydratase,for 6 h or 24 h.We harvested total RNA from the cells and performed both mRNA and miRNA array analyses,with use of Affymetrix chips,reagents,and instruments(human genome U133 plus 2.0 and miRNA 2.0 arrays).We assessed changes and their significance and interrelationships with Target Scan,Pathway Studios,and Ingenuity software.RESULTS:Changes in mRNA levels were most numerous and striking at 6 h after heme treatment but were similar and still numerous at 24 h.After 6 h of heme exposure,the increase in heme oxygenase 1 gene expression was 60-fold by mRNA and 88-fold by quantitative reverse transcription-polymerase chain reaction.We found striking changes,especially up-regulation by heme of nuclear erythroid-2 related factor-mediated oxidative stress responses,protein ubiquitination,glucocorticoid signaling,P53 signaling,and changes in RNAs that regulate intermediary metabolism.Fewer mRNAs were down-regulated by heme,and the fold decreases were less exuberant than were the increases.Notable decreases after 24 h of heme exposure were patatin-like phospholipase domain-containing protein 3(-6.5-fold),neuronal PAS domain protein 2(-1.93-fold),and protoporphyrinogen oxidase(-1.7-fold).CONCLUSION:Heme excess exhibits several toxic effects on liver and kidney,which deserve study in humans and in animal models of the human porphyrias or other disorders.展开更多
AIM:To investigate the prognostic value of vascular endothelial growth factor messenger RNA (VEGF mRNA) in the peripheral blood (PB) of patients with hepatocellular carcinoma (HCC) undergoing curative resection.METHOD...AIM:To investigate the prognostic value of vascular endothelial growth factor messenger RNA (VEGF mRNA) in the peripheral blood (PB) of patients with hepatocellular carcinoma (HCC) undergoing curative resection.METHODS:Using a reverse-transcription polymerase chain reaction (RT-PCR)-based assay, VEGF mRNA in the PB was determined prospectively in 50 controls and in 50 consecutive patients undergoing curative resection for HCC.RESULTS:Among the isoforms of VEGF mRNA, VEGF165 and VEGF121 were expressed. By multivariate analysis, a higher level of VEGF165 in preoperative PB correlated with a risk of HCC recurrence with borderline significance (P=0.050)and significantly with recurrence-related mortality (P=0.048);while VEGF121 did not. Other significant predictors of HCC recurrence included cellular dedifferentiation (P=0.033),an absent or incomplete capsule (P=0.020), vascular permeation (P=0.018), and daughter nodules (P=0.006).The other significant parameter of recurrence related mortality was cellular dedifferentiation (P=0.053). The level of circulating VEGF mRNA, however, did not significantly correlate with tumor size, cellular differentiation, capsule,daughter nodules, vascular permeation, necrosis and hemorrhage of tumors.CONCLUSION: The preoperative level of circulating VEGF mRNA, especially isoform VEGF165, plays a significant role in the prediction of postoperative recurrence of HCC.展开更多
基金Supported by A Grant from NIH/NIDDK (DK38825) to Bonkovsky HLInstitutional Funds from the Carolinas Health Care Foundation and Carolinas Medical Center
文摘AIM:To assess effects of heme on messenger RNA(mRNA) and microRNA(miRNA) profiles of liver cells derived from humans.METHODS:We exposed human hepatoma cell line Huh-7 cells to excess iron protoporphyrin(heme)(10 μmol/L) or induced heme deficiency by addition of 4,6-dioxoheptanoic acid(500 μmol/L),a potent inhibitor of aminolevulinic acid dehydratase,for 6 h or 24 h.We harvested total RNA from the cells and performed both mRNA and miRNA array analyses,with use of Affymetrix chips,reagents,and instruments(human genome U133 plus 2.0 and miRNA 2.0 arrays).We assessed changes and their significance and interrelationships with Target Scan,Pathway Studios,and Ingenuity software.RESULTS:Changes in mRNA levels were most numerous and striking at 6 h after heme treatment but were similar and still numerous at 24 h.After 6 h of heme exposure,the increase in heme oxygenase 1 gene expression was 60-fold by mRNA and 88-fold by quantitative reverse transcription-polymerase chain reaction.We found striking changes,especially up-regulation by heme of nuclear erythroid-2 related factor-mediated oxidative stress responses,protein ubiquitination,glucocorticoid signaling,P53 signaling,and changes in RNAs that regulate intermediary metabolism.Fewer mRNAs were down-regulated by heme,and the fold decreases were less exuberant than were the increases.Notable decreases after 24 h of heme exposure were patatin-like phospholipase domain-containing protein 3(-6.5-fold),neuronal PAS domain protein 2(-1.93-fold),and protoporphyrinogen oxidase(-1.7-fold).CONCLUSION:Heme excess exhibits several toxic effects on liver and kidney,which deserve study in humans and in animal models of the human porphyrias or other disorders.
文摘AIM:To investigate the prognostic value of vascular endothelial growth factor messenger RNA (VEGF mRNA) in the peripheral blood (PB) of patients with hepatocellular carcinoma (HCC) undergoing curative resection.METHODS:Using a reverse-transcription polymerase chain reaction (RT-PCR)-based assay, VEGF mRNA in the PB was determined prospectively in 50 controls and in 50 consecutive patients undergoing curative resection for HCC.RESULTS:Among the isoforms of VEGF mRNA, VEGF165 and VEGF121 were expressed. By multivariate analysis, a higher level of VEGF165 in preoperative PB correlated with a risk of HCC recurrence with borderline significance (P=0.050)and significantly with recurrence-related mortality (P=0.048);while VEGF121 did not. Other significant predictors of HCC recurrence included cellular dedifferentiation (P=0.033),an absent or incomplete capsule (P=0.020), vascular permeation (P=0.018), and daughter nodules (P=0.006).The other significant parameter of recurrence related mortality was cellular dedifferentiation (P=0.053). The level of circulating VEGF mRNA, however, did not significantly correlate with tumor size, cellular differentiation, capsule,daughter nodules, vascular permeation, necrosis and hemorrhage of tumors.CONCLUSION: The preoperative level of circulating VEGF mRNA, especially isoform VEGF165, plays a significant role in the prediction of postoperative recurrence of HCC.
