SOX7 modulates the progression of hepatoblastoma through the regulation of Wnt/ß-catenin signaling pathway.

Aims: Hepatoblastoma (HB) was reported as the frequently diagnosed primary hepatic malignant tumor among children. No reports have shown the function of SOX7 and its relationship with the Wnt/ß-catenin pathway in HB. Materials and Methods: SOX7 and factors related to Wnt/ß-catenin pathway were detected using reverse transcription-quantitative polymerase chain reaction (RT-PCR) and Western blotting. MTT 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium and flow cytometry were used to detect HB cell proliferation and apoptosis. The transwell assay uses cell invasion. Results: In this study, RT-PCR, Western blotting, and immunohistochemistry results indicated that the expression of SOX7 was significantly reduced in HB tissues compared with adjacent noncancerous tissues, while the ß-catenin was significantly increased in HB tissues compared with adjacent noncancerous tissues. There were significant differences in the PRETEXT stage and tumor metastasis between patients with low expression and high expression of SOX7. Moreover, it was found that the overexpression of SOX7 and inhibiting Wnt/ß-catenin pathway significantly reduced the cell proliferation and invasion, while the cell apoptosis was significantly increased compared with the control group. Conclusions: This study shows that SOX7 was downexpressed in HB tumor tissues. Moreover, ex vivo experiments indicated that SOX7 was related to ß-catenin and regulated the progression of HB cells.

Expression and prognostic significance of fatty acid synthase in clear cell renal cell carcinoma.

Fatty acid synthase (FASN), a key enzyme essential for fatty acid (FA) synthesis, was reportedly implicated in the initiation and progression of various cancers. However, the clinical significance of FASN in renal cell carcinoma (RCC) has not been fully elucidated yet. Here we compare the expression profile and evaluate the prognostic significance of FASN in clear cell RCC (ccRCC) patients. FASN expression was examined in 3 pairs ccRCC and their adjacent nontumor tissues by western blotting (WB) analysis, and its expression was assessed in 145 ccRCC and 13 nontumor tissues by immunohistochemistry (IHC) analysis with tissue microarrays (TMAs). The prognosis of FASN was further investigated in large-scale database using LinkedOmics (n = 537) and The Cancer Protein Atlas (TCPA, n = 445), respectively. WB detected higher FASN expression in ccRCC than normal tissues, then IHC analysis revealed that FASN expression was positively associated with histological grade, pathological stage, tumor size and metastasis status, and negatively associated with cancer-specific survival (CSS). Univariate survival analysis demonstrated that high grade, advanced stage, large tumor, metastasis, and high FASN expression were significantly associated with a shorter CSS, and multivariate analysis revealed tumor grade, stage, metastasis and FASN were identified as independent predictors for CSS in patients with ccRCC. Further LinkedOmics and TCPA analyses confirmed that high FASN expression was correlated with a poorer overall survival (OS) of ccRCC. Collectively, these findings demonstrated FASN could be a poor prognostic factor in ccRCC patients, which indicated that FA synthesis might be implicated in the tumorigenesis and progression of ccRCC.

Prognostic significance of PI3K/AKT/ mTOR signaling pathway members in clear cell renal cell carcinoma.

BACKGROUND: Renal cell carcinoma (RCC) is a fatal disease, in which the PI3K/AKT/mTOR signaling pathway serves an important role in the tumorigenesis. Previous studies have reported the prognostic significance of PI3K/AKT/mTOR signaling pathway members in RCC; however, there is insufficient evidence to date to confirm this. Thus, the present study aimed to systematically investigate the prognostic roles of multiple PI3K/AKT/mTOR signaling proteins in clear cell RCC (ccRCC) using online large-scale databases. METHODS: The mRNA expression profiles of PI3K/AKT/mTOR signaling pathway proteins PTEN, PIK3CA, PIK3CB, PIK3CD, PIK3CG, AKT1, AKT2, AKT3 and mTOR were investigated using the Gene Expression Profiling Interactive Analysis (GEPIA) and Oncomine databases, and the protein expression levels of PI3K, AKT and mTOR were detected using western blotting (WB) analysis. In addition, the correlation between mRNA or protein expression levels and the prognostic significance was analyzed using the Kaplan-Meier (K-M) plotter (n = 530), the Human Protein Atlas (HPA; n = 528) and The Cancer Protein Atlas (TCPA; n = 445) databases. RESULTS: The GEPIA revealed that the mRNA expression of major PI3K/AKT/mTOR pathway members, including PTEN, PIK3CA, PIK3CB, AKT1, AKT2 and AKT3, were negatively correlated with ccRCC stages (P < 0.05), though most of their mRNA and protein expression levels were notsignificantly different between ccRCC and normal tissues using GEPIA, Oncomine and WB analyses (P < 0.05). Meanwhile, using the K-M plotter and HPA prognostic analysis, it was found that the mRNA expression levels of the majority of the PI3K/AKT/mTOR signaling pathway members, including PTEN, PIK3CA, PIK3CB, PIK3CG, AKT3 and mTOR were positively correlated with overall survival (OS), whereas PIK3CD mRNA expression was negatively correlated with OS (P < 0.05). Furthermore, TCPA prognostic analysis observed that several of the key molecules of the PI3K/AKT/mTOR signaling pathway [PTEN, p-AKT (S473) and p-mTOR (S2448)] were also positively correlated with OS in patients with ccRCC (P < 0.05). In conclusion, the present study suggested that several members of the PI3K/AKT/mTOR signaling pathway, especially PTEN, may be favorable prognostic factors in ccRCC, which indicated that the PI3K/AKT/mTOR signaling pathway may be implicated in ccRCC initiation and progression.

The mRNA Expression Signature and Prognostic Analysis of Multiple Fatty Acid Metabolic Enzymes in Clear Cell Renal Cell Carcinoma.

Renal cell carcinoma (RCC) is a metabolic disease, and accumulating evidences indicate significant alterations in the cellular metabolism, especial aerobic glycolysis and glutamine metabolism, in RCC. However, fatty acid (FA) metabolism has received less attention, and the mRNA expression pattern and prognostic role of FA metabolic enzymes in clear cell RCC (ccRCC) have not been carefully examined. In the current study, we first investigated the mRNA expression profiles of multiple FA metabolic enzymes, i.e., ACLY, ACC, FASN, SCD, CPT1A, HADHA, HADHB, and ACAT1, in 42 ccRCC and 33 normal kidney tissues using the Oncomine database, validated their mRNA expression profiles using GEPIA resource, then evaluated and validated the prognostic significance of these metabolic enzymes in 530 ccRCC patients using Kaplan-Meier plotter and GEPIA analyses respectively. The Oncomine and GEPIA confirmed higher ACLY, SCD, and lower ACAT1 mRNA expression in ccRCC than normal tissues (P<0.05). And further prognostic analysis displayed that overexpression of the some FA anabolic enzymes (FASN) was correlated to poor overall survival (OS), while overexpression of the FA catabolic enzymes (CPT1A, HADHA, HADHB, and ACAT1) was correlated to favorable OS in ccRCC patients. In conclusion, multiple FA metabolic enzymes, such as FASN, HADHA, and ACAT1, were potential prognostic markers of ccRCC, which implied alterations in FA metabolism might be involved in ccRCC tumorigenesis and progression.

HADHA overexpression disrupts lipid metabolism and inhibits tumor growth in clear cell renal cell carcinoma.

Hydroxyacyl-CoA dehydrogenase alpha subunit (HADHA) is a key lipid metabolic enzyme with a novel role in carcinogenesis. We previously reported that HADHA, a prognostic marker, was downregulated in clear cell renal cell carcinoma (ccRCC). Herein, the tumor inhibitory role of HADHA overexpression in ccRCC was investigated further. The quantitative proteomic analysis displayed that a total of 1293 and 1293 proteins were identified in HADHA overexpressed 786-O-hadha and vector-transfected control 786-O-vc cells, respectively, and 206 proteins were found to be up- or downregulated. PANTHER, OmicsNet, STRING, and DAVID tools were utilized on the dysregulated proteins in order to elucidate multiple metabolic pathways (especial lipid metabolism) and lipid metabolism-related proteins (e.g. ACAT1, ACLY). The dysregulation of the lipid metabolic enzymes, ACAT1, ACLY, CYB5R3 and FASN, were confirmed by Western blotting. Further assays demonstrated that HADHA overexpression significantly inhibited cell growth, induced cell apoptosis, and decreased the formation of cytoplasmic lipid droplets (LDs); moreover, it also inhibited tumor growth and lessened the formation of LDs in xenografted mouse. Collectively, these data revealed that HADHA overexpression disrupted lipid metabolism and inhibited tumor growth, which shed light on HADHA as a potential therapeutic target for clinical intervention of ccRCC.

Evaluation and prognostic significance of manganese superoxide dismutase in clear cell renal cell carcinoma.

The antioxidant enzyme manganese superoxide dismutase (MnSOD) is up-regulated in renal cell carcinoma (RCC) and has been implicated in multiple stages of RCC tumorigenesis and progression. However, the prognostic significance of MnSOD in RCC has not been fully elucidated. This study aimed to investigate the expression profile of MnSOD in clear cell RCC (ccRCC) tissues and evaluate the clinical significance of this enzyme in ccRCC patients. MnSOD mRNA was assessed in 42 ccRCC and 33 normal kidney tissues using the Oncomine database, and its protein was detected in 145 ccRCCs and 3 normal tissues by immunohistochemistry staining. The Oncomine database confirmed higher MnSOD mRNA expression in ccRCC than in normal tissues, and immunohistochemistry analysis revealed that MnSOD protein expression was inversely associated with pathologic grade, clinical stage, tumor size, M status, and cancer-specific survival. In addition, univariate survival analysis demonstrated that high-grade, late-stage, large tumors, stage M1, and low MnSOD expression were associated with a poorer prognosis for cancer-specific survival, and further multivariate analysis revealed that tumor grade, stage, M1 stage, and MnSOD were identified as independent prognostic factors for cancer-specific survival in patients with ccRCC. Collectively, these findings imply that MnSOD is a promising prognostic marker in ccRCC and implies that oxidative stress might be involved in the tumorigenesis and progression of ccRCC.

Exosomal miR-34s panel as potential novel diagnostic and prognostic biomarker in patients with hepatoblastoma.

PURPOSE: The aim of this study is to identify the diagnostic values of serum exosomal miRNA-34s of patients with HB in a large Asian group and explore the prognostic value of the exosomal miRNA-34s panel compared with other risk factors. METHODS: We retrospectively reviewed 89 children with HB. Among these patients, 63 patients were included as training group to build the diagnostic model for HB. 26 patients were defined as the validation group. The expressions of miRNA-34s were detected by real-time PCR. The comparison of diagnostic and prognostic performance of serum exosomal miRNA-34s was measured using the area under ROC curve (AUC). RESULTS: For patients in the training group, expression of miRNA-34a, miRNA-34b and miRNA-34c was significantly lower in patients with HB compared with control group in serum exosomes. Between HB training group and the control group, exosomal miRNA-34a, miRNA-34b and miRNA-34c had no significant differences compared with the AFP level in diagnosing HB. The performance of the exosomal miRNA-34s panel in differentiating the HB training group from the control group was superior to the AFP level. The value of the exosomal miRNA-34s panel in predicting prognosis of patients with HB was superior to other risk factors in both training group and validation group. CONCLUSIONS: In this study, we found that the expression of exosomal miRNA-34a, miRNA-34b and miRNA-34c was significantly lower in patients with HB compared with the control group, and we confirmed the exosomal miRNA-34s panel could be defined as a diagnostic and prognostic biomarker for patients with HB. LEVEL OF EVIDENCE: Level II. TYPE OF STUDY: Retrospective Study.

Combined low miR-34s are associated with unfavorable prognosis in children with hepatoblastoma: A Chinese population-based study.

PURPOSE: The aim of this study is to identify the association between miR-34's family and the prognosis of HB in a large Asian cohort and to explore the interaction of miR-34 with other independent risk factors in the process of affecting prognosis of HB. METHODS: We retrospectively reviewed 78 children with HB (36 female, 42 male) managed in our institutions between 2007 and 2014. The expression of miR-34 was detected by real-time PCR. Prognostic factors were evaluated using Kaplan-Meier curves and Cox proportional hazards models. RESULTS: For the entire cohort of 76 patients, The normalized real-time PCR results showed that all three miRNAs were deregulated in tumor tissues as compared with corresponding noncancerous tissue samples. Descriptive survival statistics and Kaplan-Meier curves suggested that AFP levels, metastases, vascular invasion, PRETEXT stage and miR-34 had prognostic significance in this relatively selected cohort. After that we made miR-34 into different combinations. The results demonstrated that combined low miR-34a and miR-34b (HR:2.212, P=0.016), combined low miR-34a and miR-34c (HR:1.984, P=0.025) and combined low miR-34a, miR-34b and miR-34c (HR:3.569, P=0.001) were independent prognostic factors of HB. We further conduct stratified analysis of the impact of other identified risk factors on the combined low of three miR-34. CONCLUSIONS: In this study, we found that miR-34s were deregulated in tumor tissues compared with corresponding noncancerous tissue samples. We also confirmed that combined low miR-34 is an independent prognostic factor related with HB.

Pure fetal histology subtype was associated with better prognosis of children with hepatoblastoma: A Chinese population-based study.

BACKGROUND AND AIM: The aim of this study is to identify the association between histologic types and the prognosis of hepatoblastoma (HB) in a large Asian cohort of a single institution and to explore the interaction of histologic types with other independently risk factors in the process of affecting prognosis of HB. METHODS: We retrospectively reviewed 176 children with HB (82 female, 94 male) managed in our institution between May 1, 2001 and July 30, 2014. Prognostic factors were evaluated using Kaplan-Meier curves and Cox proportional hazards models. RESULTS: For the entire cohort of 176 patients, the overall median survival was 80.4 months(95% CI: 71.6-89.2 months), and the 5-year event-free survival and overall survival rates were 54.6 and 66.7%. Descriptive survival statistics and Kaplan-Meier curves suggested that alpha fetoprotein levels, tumor metastases, multifocality, histologic types, and Pre-Treatment Extent of Disease staging System stage had prognostic significance in this relatively selected cohort. Moreover, after eliminating the impact of the interaction of different classification methods of histologic types, pure fetal histologic (PFH) was an independent prognostic factor of HB (hazard ratio [HR]: 2.752, P = 0.021). Further stratification analysis showed that the impaction of other identified risk factors on the influence of PFH on the prognosis of HB patients was different. CONCLUSIONS: We have confirmed that the HB prognostic factors of HB and PFH was associated with better prognosis of children with HB based on an Asian population. PFH showed different significance in the process of affecting prognosis of HB with the interaction of other independent risk factors.

Conditional probability of survival in patients with biliary atresia after Kasai portoenterostomy: a Chinese population-based study.

Survival estimates are commonly reported for the outcomes of patients. Conditional probability, which offers more relevant information regarding the prognosis for patients once they have survived for some time, are lacking for patients with biliary atresia. The aim of this study is to calculate the conditional probability of survival in patients with biliary atresia after Kasai portoenterostomy. Clinical data from 244 consecutive patients who underwent Kasai portoenterostomy were reviewed and the actuarial survival estimated. Conditional survival probabilities and 95% CIs were calculated. Of the 244 patients, the overall median NLS was 41.2 months (95% CI:30.8-51.6 months), and the 1-year, 3-year, and 5-year NLS rates were 85.4%, 61.1%, and 43.3%, respectively. The probabilities of surviving at the time point of 5 years given survival to 1, 2, 3, and 4 years were 50%, 56%, 73%, and 93%, respectively. For conditional survival for 1 additional year, patients with age >90 days and cholangitis had significantly predictive values(both P<0.05). Cox proportional hazards model showed that age > 90 days(HR: 2.519, P=0.001), and postoperative cholangitis(HR:2.568, P<0.001) were associated with an adverse NLS in the multivariable adjusted analysis. our findings showed that conditional survival of patients with BA is not linear over time after Kasai operation. Information derived from conditional survival can be used to better manage patients with BA, including the potential subsequent treatment decisions, the chance of listing for transplantation, or even the end-of-life planning.