Aberrant expression of peroxiredoxin 1 and its clinical implications in liver cancer

AIM: To investigate the expression characteristics of peroxiredoxin 1(PRDX1) m RNA and protein in liver cancer cell lines and tissues.METHODS: The RNA sequencing data from 374 patients with liver cancer were obtained from The Cancer Genome Atlas. The expression and clinical characteristics of PRDX1 m RNA were analyzed in this dataset. The Kaplan-Meier and Cox regression survival analysis was performed to determine the relationship between PRDX1 levels and patient survival. Subcellular fractionation and Western blotting were used to demonstrate the expression of PRDX1 protein in six liver cancer cell lines and 29 paired fresh tissue specimens. After bioinformatics prediction,a putative posttranslational modification form of PRDX1 was observed using immunofluorescence under confocal microscopy and immunoprecipitation analysis in liver cancer cells.RESULTS: The m RNA of PRDX1 gene was upregulated about 1.3-fold in tumor tissue compared with the adjacent non-tumor control(P = 0.005). Its abundance was significantly higher in men than women(P < 0.001). High levels of PRDX1 m RNA were associated with a shorter overall survival time(P =0.04) but not with recurrence-free survival. The Cox regression analysis demonstrated that patients with high PRDX1 m RNA showed about 1.9-fold increase of risk for death(P = 0.03). In liver cancer cells,PRDX1 protein was strongly expressed with multiple different bands. PRDX1 in the cytosol fraction existed near the theoretical molecular weight,whereas two higher molecular weight bands were present in the membrane/organelle and nuclear fractions. Importantly,the theoretical PRDX1 band was increased,whereas the high molecular weight form was decreased in tumor tissues. Subsequent experiments revealed that the high molecular weight bands of PRDX1 might result from the post-translational modification by small ubiquitin-like modifier-1(SUMO1).CONCLUSION: PRDX1 was overexpressed in the tumor tissues of liver cancer and served as an independent poor prognostic factor for overall survival. PRDX1 can be modified by SUMO to play specific roles in hepatocarcinogenesis.

Establishment of a Sandwich ELISA Method for Detection of Vascular Endothelial Growth Factor in Serum Samples of Hepatocellular Carcinoma Patients

Objective To establish a sandwich ELISA method for detecting vascular endothelial growth factor （VEGF） in sera of population and the patients with hepatocellular carcinoma （HCC）. Methods Full length and two truncated human VEGF cDNA sequences were amplified from a commercial plasmid pBLAST49-hVEGF by PCR and inserted into the prokaryotic-expression plasmid pET-32a or pGEX-2T. Various VEGF proteins were expressed and purified from E. coli in His-Trx or GST fusion forms. The specific VEGF antibodies were elicited in experimental rabbits and mice by immunization of the full length VEGF fusion protein His-Trx-VEGF1-165. After purification of antibodies with chromatograph of Protein G, a sandwich ELISA technique was established. Serum VEGF levels were evaluated in 229 adults and 291 HCC patients. Results SDS-PAGE displayed that the molecular weights of the expressed full length （His-Trx-VEGF1-165）, N-terminal （His-Trx-VEGF1-100） and C-terminal （GST-VEGF100-165） human VEGF fusion proteins were about 38KD, 31KD, and 33KD, respectively. Western blots confirmed that the prepared antisera were able to recognize both prokaryoticly and eukaryoticly expressed recombinant VEGF proteins. Assays of serially diluted His-Trx-VEGF1-100 by the established sandwich ELISA method showed that the linear range of the standard curve was 0.625-320 ng/mL, with the squared correlation coefficient R^2=0.991. Screening of a serum panel containing 291 serum samples of HCC patients and 229 health adults revealed that the average VEGF level in HCC patients was higher than that in healthy controls, with a statically significant difference. Conclusion The established sandwich ELISA reflects the level of serum VEGF and provide scientific basis for screening metastasis and recurrence of HCC using serum VEGF as an index.

Clinicopathological and Immunohistochemical Study of Low-Grade Myofibroblastic Sarcoma of the Liver One Case Report

Low-grade myofibroblastic sarcoma is a recently characterized tumor showing features of myofibroblastic differen-tiation that is part of the spectrum of malignant mesenchymal tumors. This extremely rare type of tumor occurs most commonly in superficial locations. The case we describe herein is that of a 60- year-old man with two large hepatic masses. The patient＇s tumor was removed radically through an incision due to the inconclusive imaging findings. Follow-up computed tomography showed no recurrence and metastasis after 37 months, suggesting that enucleation was adequate for tumor eradication. To our knowl-edge, this is the first reported case of low-grade myofibroblastic sarcoma in the liver in the English language literature.

Excision Repair Cross-complementation Group 1 is a Prognostic Biomarker in Patients with Colorectal Cancer Receiving Chemotherapy

Background：Conflicting results about the association between expression level of excision repair cross-complementation group 1 （ERCC1） and clinical outcome in patients with colorectal cancer （CRC） receiving chemotherapy have been reported.Thus,we searched the available articles and performed the meta-analysis to elucidate the prognostic role of ERCC1 expression in patients with CRC.Methods：A thorough literature search using PubMed （Medline）,Embase,Cochrane Library,Web of Science databases,and Chinese Science Citation Database was conducted to obtain the relevant studies.Pooled hazard ratios （HRs） or odds ratios （ORs） with 95% confidence intervals （CIs） were calculated to estimate the results.Results：A total of 11 studies were finally enrolled in this meta-analysis.Compared with patients with lower ERCC1 expression,patients with higher ERCC1 expression tended to have unfavorable overall survival （OS） （HR =2.325,95% CI：1.720-3.143,P 〈 0.001）,progression-free survival （PFS） （HR =1.917,95% CI：1.366-2.691,P 〈 0.001） and poor response to chemotherapy （OR =0.491,95% CI：0.243-0.990,P =0.047）.Subgroup analyses by treatment setting,ethnicity,HR extraction,detection methods,survival analysis,and study design demonstrated that our results were robust.Conclusions：ERCC1 expression may be taken as an effective prognostic factor predicting the response to chemotherapy,OS,and PFS.Further studies with better study design and longer follow-up are warranted in order to gain a deeper understanding of ERCC 1＇s prognostic value.

Evaluation of Eight Different Clinical Staging Systems Associated with Overall Survival of Chinese Patients with Hepatocellular Carcinoma

Background： Hepatocellular carcinoma （HCC） is a common cancer in China, an area of high hepatitis B virus （HBV） infection. Although several staging systems are available, there is no consensus on the best classification to use because multiple factors, such as etiology, clinical treatment and populations could affect the survival of HCC patients. Methods： This study analyzed 743 HBV-related Chinese HCC patients who received surgery first and evaluated the predictive values of eight different commonly used staging systems in the clinic. Results： The overall 1-, 3-, 5-year survival rates and a median survival were 91.5%, 70.3%, 55.3% and 72 months respectively. Barcelona Clinic Liver Cancer （BCLC） staging systems had the best stratification ability and showed the lowest Akaike information criterion （AIC） values （2896.577）, followed by tumor-node-metastasis Th （TNM 7^th） （AIC = 2899.980）, TNM 6th （AIC = 2902.17）, Japan integrated staging score （A1C = 2918.085）, Tokyo （AIC = 2938.822）, Cancer of the Liver Italian Program score （AIC = 2941.950）, Chinese University Prognostic Index grade （AIC = 2962.027）, and Okuda （AIC = 2979.389）. Conclusions： BCLC staging system is a better staging model for HBV infection patients with HCC in Chinese population among the eight currently used staging systems. These identifications afford a large group of Chinese HCC patients with HBV infection and could be helpful to design a new staging system for a certain population.

Prognostic Analysis of 102 Patients with Synchronous Colorectal Cancer and Liver Metastases Treated with Simultaneous Resection

Background： The liver is the most common site for colorectal cancer （CRC） metastases. Their removal is a critical and challenging aspect of CRC treatment. We investigated the prognosis and risk factors of patients with CRC and liver metastases （CRCLM） who underwent simultaneous resections for both lesions. Methods： From January 2009 to August 2016, 102 patients with CRCLM received simultaneous resections of CRCLM at our hospital. We retrospectively analyzed their clinical data and analyzed their outcomes. Overall survival （OS） and disease-free survival （DFS） were examined by Kaplan-Meier and log-rank methods. Results： Median follow-up time was 22.7 months： no perioperative death or serious complications were observed. Median OS was 55.5 months： postoperative OS rates were l-year： 93.8%, 3-year： 60.7%, and 5-year： 46.4%. Median DFS was 9.0 months; postoperative DFS rates were l-year： 43.1%, 3-year： 23.0%, and 5-year 21.1%. Independent risk factors found in multivariate analysis included carcinoembryonic antigen 〉100 ng/ml, no adjuvant chemotherapy, tumor thrombus in liver metastases, and bilobar liver metastases for OS; age 〉60 years, no adjuvant chenlotherapy, multiple metastases, and largest diameter ≥3 cm for DFS. Conclusions： Simultaneous surgical resection is a safe and effective treatment for patients with synchronous CRC LM. The main prognostic factors are pathological characteristics of liver metastases and whether standard adjuvant chemotherapy is performed.