Identification of potential biomarkers for diagnosis of hepatocellular carcinoma.

Hepatocellular carcinoma (HCC) has a high mortality rate owing to its complexity. Identification of abnormally expressed genes in HCC tissues compared to those in normal liver tissues is a viable strategy for investigating the mechanisms of HCC tumorigenesis and progression as a means of developing novel treatments. A significant advantage of the Gene Expression Omnibus (GEO) and The Cancer Genome Atlas (TCGA) is that the data therein were collected from different independent researchers and may be integrated, allowing for a more robust data analysis. Accordingly, in the present study, the gene expression profiles for HCC and control samples were downloaded from the GEO and TCGA. Functional enrichment analysis was performed using a Metascape dataset, and a protein-protein interaction (PPI) network was constructed using the Search Tool for the Retrieval of Interacting Genes/proteins (STRING) online database. The prognostic value of mRNA for HCC was assessed using the Kaplan-Meier Plotter, a public online tool. A gene mRNA heatmap and DNA amplification numbers were obtained from cBioPortal. A total of 2,553 upregulated genes were identified. Functional enrichment analysis revealed that these differentially expressed genes (DEGs) were mainly accumulated in metabolism of RNA and the cell cycle. Considering the complexity and heterogeneity of the molecular alterations in HCC, multiple genes for the prognostication of patients with HCC are more reliable than a single gene. Thus, the PPI network and univariate Cox regression analysis were applied to screen candidate genes (small nuclear ribonucleoprotein polypeptide B and B1, nucleoporin 37, Rac GTPase activating protein 1, kinesin family member 20A, minichromosome maintenance 10 replication initiation factor, ubiquitin conjugating enzyme E2 C and hyaluronan mediated motility receptor) that are associated with the overall survival and progression-free survival of patients with HCC. In conclusion, the present study identified a set of genes that are associated with overall survival and progression-free survival of patients with HCC, providing valuable information for the prognosis of HCC.

Independent and Joint Associations between Serum Calcium, 25-Hydroxy Vitamin D, and the Risk of Primary Liver Cancer: A Prospective Nested Case-Control Study.

BACKGROUND: Accumulating evidence has shown that serum calcium and vitamin D may be associated with or influence various cancer risks. However, no prospective studies have evaluated the independent and joint associations between prediagnostic levels of serum calcium and vitamin D and future risk of incident primary liver cancer. METHODS: We used a nested case-control design to evaluate subjects over 22 years of follow-up. Serum calcium, 25-hydroxy vitamin D [25(OH)D], and three markers of hepatitis B virus and hepatitis C virus were measured in baseline serum from 226 incident primary liver cancer cases and 1,061 matched controls. We calculated ORs and 95% confidence intervals (CI) using logistic regression to estimate the associations between calcium, 25(OH)D, and primary liver cancer risk. RESULTS: Multivariable adjusted models showed that subjects with both low (ORLow/Medium = 1.48, 95% CI = 1.01-2.17) or high (ORHigh/Medium = 1.92, 95% CI = 1.34-2.76) calcium had an increased primary liver cancer risk, while those with high 25(OH)D had a decreased risk of primary liver cancer (ORHigh/Medium = 0.54, 95% CI = 0.35-0.82). In joint analyses, when compared with subjects with medium calcium and 25(OH)D, subjects with high calcium and medium 25(OH)D had elevated odds of developing primary liver cancer (OR = 1.89, 95% CI = 1.17-3.05); those with medium calcium and high 25(OH)D had reduced odds of developing primary liver cancer (OR = 0.34, 95% CI = 0.17-0.67); and subjects in other classifications of calcium and serum 25(OH)D levels had no change in the odds of developing primary liver cancer (all P > 0.05). CONCLUSIONS: In a nutrient-deficient population, we found that serum calcium and serum 25(OH)D could potentially be modifiable risk or protective factors. IMPACT: Our findings provide potential targets for primary liver cancer prevention and control.

MAPRE1 promotes cell cycle progression of hepatocellular carcinoma cells by interacting with CDK2.

Targeting cyclin-dependent kinases (CDKs) is a promising method of therapy for cancer. Unfortunately, the efficacy of CDK inhibitors in hepatocellular carcinoma (HCC) is limited, due in part to incomplete understanding of cell cycle progression and a lack of specific biomarkers to adequately identify which patients may be responsive to CDK inhibitors. In the present study, we report that microtubule-associated protein RP/EB family member 1 (MAPRE1), a gene involved in cell cycle and microtubule regulation, is significantly increased in HCC tissue, promotes HCC cell proliferation, enhances in vitro tumorigenesis, and associates with poor prognosis of HCC. We demonstrate that MAPRE1 binds with CDK2, resulting in the hyperphosphorylation of the CDK2 Thr161 residue in HCC cells. Our findings reveal that targeting MAPRE1 might be an effective therapeutic strategy in HCC, and suggest that MAPRE1 expression might provide a promising biomarker to stratify patients with HCC who may benefit from treatment with CDK inhibitors.

FBXW10 promotes hepatocarcinogenesis in male patients and mice.

Hepatocellular carcinoma (HCC) is reported to associate with abnormal expression of SCF E3 ubiquitin ligases. FBXW10, an F-box protein of the E3 ubiquitin ligases, was abnormally regulated in HCC patients. However, whether FBXW10 is associated with HCC has not yet been evaluated. Here, we analyzed the associations between overall survival and various risk factors in 191 HCC tissues. Univariate and multivariate analyses demonstrated that FBXW10 was an independent risk factor related to HCC prognosis. The results showed that FBXW10, gender and tumor state were strongly associated with overall survival in HCC patients. Furthermore, high expression of FBXW10 was associated with poor survival among male HCC patients but not female HCC patients. FBXW10 was more highly expressed in male HCC tissues and more strongly related to vascular invasion in male HCC patients. Consistent with these findings, the male FBXW10-Tg(+) mice were more susceptible to tumorigenesis, changes in regenerative capacity, and liver injury and inflammation but not changes in liver function than FBXW10-Tg(-) mice. FBXW10 promoted cell proliferation and migration in HCC cell lines. Our findings reveal that FBXW10, an independent risk factor for HCC, promotes hepatocarcinogenesis in male patients, and is also a potential prognostic marker in male patients with HCC.

Association between C-reactive protein, incident liver cancer, and chronic liver disease mortality in the Linxian Nutrition Intervention Trials: a nested case-control study.

BACKGROUND: C-reactive protein (CRP) is a marker of systemic inflammation that has been associated with the incidence and prognosis for a number of different cancers. Recent data suggest that CRP may be a prognostic factor for liver cancer and cirrhosis. However, few long-term studies are available. METHODS: We prospectively examined associations between serum CRP and subsequent risk of liver cancer incidence or chronic liver disease mortality in a nested case-control study performed in the Linxian Nutrition Intervention Trials cohort. Baseline serum CRP was measured for 220 incident liver cancer cases, 276 participants who died of chronic liver disease, and 1,018 age-, sex-, and trial-matched controls. Unconditional logistical regression models were used to estimate ORs and 95% confidence intervals (CI). RESULTS: Compared with the lowest quartile, subjects in the fourth quartile of serum CRP had a higher risk of liver cancer incidence (OR, 1.63; 95% CI, 1.06-2.51), with a significant Ptrend across quartiles (P = 0.01). The association with liver cancer was only significant among men (Q4 vs. Q1; OR, 2.00; 1.10-3.62), but not among women (Q4 vs. Q1; OR, 1.15; 0.60-2.22). For chronic liver disease deaths, the corresponding risk estimate in men and women was 2.95 (1.90-4.57), with a monotonic trend (P < 0.001). CONCLUSIONS: Higher serum CRP concentrations at baseline were associated with subsequent incidence of liver cancer and death from chronic liver disease. IMPACT: Our findings suggest that levels of systemic inflammation may serve as a long-term marker of liver cancer and liver disease.

Multivitamin and mineral supplementation is associated with the reduction of fracture risk and hospitalization rate in Chinese adult males: a randomized controlled study.

Controversy exists in the literature regarding the efficacy of bone health-related nutrients, especially calcium and vitamin D, in preventing fractures. The aim of our present study was to determine the effect of multivitamin and mineral supplementation on fracture incidence among 3,318 participants from a nutritional intervention trial in Linxian, China. A total of 1,461 men and 1,857 women were enrolled and randomized to daily supplementation with 26 vitamins and minerals tablet or placebo pills for 6 years, followed by a 16-year post-interventional follow-up. The dates, sites, and causes of the fractures were collected retrospectively via a standardized questionnaire. Cox proportional hazard model was used to estimate hazard ratios and 95% confidence intervals of fracture incidence in the intervention versus the placebo group. A total of 221 fractures (57 in men and 164 in women) occurred during the entire study period of 21 years and 9 months. In men, the supplement reduced the risk of fracture by 63% during the trial period, and this protective effect was sustained and statistically significant when analysis included both the trial period and 5- or 10-year post-intervention follow-up (years 0-11, P = 0.04; years 0-16, P = 0.02, respectively). The protection against fracture was not apparent >10 years after cessation of the intervention. In women, no significant effect of supplementation on fracture incidence was seen in any of the study periods. These results demonstrate that a 6-year multivitamin and mineral intervention was associated with significant reduction of fracture risk and fracture-related hospitalization in men, but not in women.

[Association between serum calcium levels and the risk of liver cirrhosis].

OBJECTIVE: To examine the association between serum calcium levels and the risk of liver cirrhosis. METHODS: A nested case-control study was performed based on the nutritional intervention trial of esophageal carcinoma in Linxian, Henan province. Serum samples of 281 liver cirrhosis cases and 562 controls were tested for calcium concentrations, surface antigen (HBsAg) and core antibody (anti-HBc) on Hepatitis virus B and antibody on Hepatitis virus C (anti-HCV), using automatic serum biochemical analysis system and enzyme-linked immunoassay. Data on baseline characteristics were collected via a questionnaire. Serum calcium value was divided into tertiles and logistic regression model was used to calculate odds ratios and 95% confidence intervals. RESULTS: Individuals in the case group showed higher calcium levels (1.81 ± 0.84) mmol/L when compared to the controls (1.65 ± 0.79) mmol/L, with t = -2.640 and P = 0.008. The calcium levels were associated with the risk of liver cirrhosis (χ(2) = 6.888, P = 0.0319). Risks for the individuals in the highest tertile were doubled when compared to the lowest (adjusted OR = 2.261, 95%CI: 1.497 - 3.416, P = 0.002). A positive correlation was observed between serum calcium level and the risk of liver cirrhosis (χ(2) = 6.842, P = 0.0089). CONCLUSION: from our study revealed that the elevated serum calcium level might be an independent risk factor for liver cirrhosis. However, further investigations are anticipated to explore its potential mechanisms.

[Prospective study on the relation between serum vitamin D levels and liver cirrhosis risk].

OBJECTIVE: To explore the relation between serum vitamin D (25(OH)D) concentrations and liver cirrhosis. METHODS: A nested case control study was designed based on the Nutrition Intervention Trial (NIT) cohort, from which non-degraded serum samples and complete baseline and follow-up data were available for 282 individuals diagnosed with liver cirrhosis and 564 healthy controls. The serum samples were tested by enzyme-linked immunosorbent assay to detect and quantify 25(OH)D, as well as hepatitis B virus surface antigen and core antibody and hepatitis C virus antibody. The study participants were divided into four groups according to quartile range of 25(OH)D concentration and logistic regression modeling was used to evaluate the relation with liver cirrhosis risk by estimating odds ratios (ORs) and 95% confidence intervals (CIs). RESULTS: The liver cirrhosis group had a significantly lower concentration of 25(OH)D than the control group (median and quartile range in nmol/L, cases: 15.3 and 13.8; controls: 20.2 and 16.6; t = 5.169, P = 0.002). When the first interval was set as the reference level, the ORs(95% CIs) of the second, third, and fourth intervals were: 0.495(0.333 - 0.736), 0.452(0.303 - 0.675), and 0.349(0.231- 0.527). After adjusting the analysis for the presence of pathogenic-related confounding factors, the ORs(95% CIs) of the second, third, and fourth intervals were: 0.499(0.328 - 0.759), 0.427(0.278 - 0.654), and 0.344(0.222-0.532). The 25(OH)D level was inversely correlated with risk of liver cirrhosis (Chi2 = 25.200, P < 0.001). CONCLUSION: Risk of liver cirrhosis increases as 25(OH)D serum concentration decreases. Vitamin D might function as a protective factor against development of cirrhosis.

[Prospective study of the association between fasting glucose concentrations and liver cirrhosis risk].

OBJECTIVE: To investigate the association between concentration levels of fasting serum glucose and liver cirrhosis. METHODS: A nested case-control study was carried out based on the sample cohort from the Nutrition Intervention Trials previously conducted in one country in Henan province. Using an automatic biochemical analysis system and enzyme-linked immunoassay, baseline serum samples from 310 liver cirrhosis patients and 620 healthy controls were tested for fasting glucose concentration, hepatitis B surface antigen (HBsAg), hepatitis B core antibody (anti-HBc), and hepatitis C virus antibody (anti-HCV). Baseline demographic information was collected by questionnaire. The serum glucose values were divided into quintiles and applied to a logistic regression model to estimate odds ratios (ORs) and 95% confidence intervals (95% CIs). RESULTS: The mean fasting blood glucose level was significantly higher in cases (4.5+/-1.8 mmol/L) than in controls (4.2+/-2.1 mmol/L) (t=-2.414, P=0.016). The individuals in the highest quintile had a significantly higher risk of disease than those in the lowest quintile [OR=1.672 (1.080, 2.588)]. Moreover, increase in glucose level was accompanied by increased risk, and the relation showed statistically significant linearity (P=0.002). The statistical significance of risk remained after adjustment for potential confounders, including sex, age, HBsAg, anti-HBc, and residence running water status [OR=1.96 (1.216, 3.157), P=0.001]. CONCLUSION: Elevated serum fasting glucose concentration was an independent risk factor of cirrhosis.