Serum TSGF and miR-214 levels in patients with hepatocellular carcinoma and their predictive value for the curative effect of transcatheter arterial chemoembolization.

BACKGROUND: The purpose of this study is to evaluate the expression of tumor-specific growth factor (TSGF) and microRNA-214 (miR-214) in the serum of patients with primary hepatocellular carcinoma (PHC) and their predictive values for the curative effect of transcatheter arterial chemoembolization (TACE). METHODS: A retrospective analysis of the clinical data of 87 PHC patients were treated with TACE. According to the curative effect 1 month after TACE, PHC patients were divided into disease control group (n=56) and disease progression group (n=31). The expression levels of TSGF and miR-214 were detected by qRT-PCR before or after treatment with TACE in disease control group and disease progression group. The predictive value of TSGF and miR-214 for the efficacy of TACE were evaluated by the receiver operating characteristic (ROC) curve. The Kaplan-Meier survival curve was drawn according to the critical value of ROC. The effect of pretreatment levels of TSGF and miR-214 was analyzed on the three-year survival rate of patients. RESULTS: After TACE treatment, the mRNA expressive of serum TSGF were increased in progression group, while the levels of miR-214 were decreased compared with control group (P<0.05). Importantly, at one month after TACE treatment, the levels of TSGF were decreased in control group and progression group, while the expression of miR-214 was increased compared with in both groups before TACE treatment(P<0.05). The result of ROC analysis showed that for predicting the curative effect of TACE, the levels of TSGF and miR-214 expression were (181.32, 0.63) (0.849 and 0.807) and (0.759-0.938, 0.707-0.907) in the cutoff values, AUCs and 95% confidence intervals, respectively. The result of Kaplan-Meier analysis showed that the 3-year survival rate of the low TSGF group was up-regulated than that of the high TSGF group [65.12% (31/44) vs. 30.23% (13/43); χ2=5.014; P=0.025]. The 3-year survival rate of the miR-214 high expression group was up-regulated than that of the miR-214 low expression group [61.70% (29/47) vs. 30.00% (12/40); χ2=6.928; P=0.008]. CONCLUSIONS: Serum TSGF and miR-214 could be used as potential biomarkers for PHC diagnosis and prognosis.

Revealing A-Raf functions through its interactome.

A-Raf is a member of the Raf kinase family. Unlike B-Raf and C-Raf, the functions of A-Raf remain obscure. To gain more insight into the biological functions of A-Raf, we investigated the A-Raf interactome using proteomics. We found 132 proteins that interact with A-Raf and confirmed the interaction of 12 of these proteins with A-Raf by western blotting. Our data suggested that A-Raf regulates apoptosis, RNA catabolism, GTPase activity, and cell adhesion by interacting with proteins located in different cellular compartments. We identified all ten hallmarks of cancer in these interacting proteins, suggesting that A-Raf is involved in carcinogenesis. Our results also indicated that A-Raf may play a role in different diseases and signaling pathways. These findings have identified potential regulators of A-Raf and provide a systemic insight into its biological functions.

Decoding the full picture of Raf1 function based on its interacting proteins.

Raf1 is a member of the Raf kinase family and regulates many fundamental cell processes, including proliferation, differentiation, apoptosis, motility, and metabolism. However, the functions of Raf1 have not been completely elucidated. To better understand Raf1 function, we investigated the proteins that interacted with Raf1. We identified 198 Raf1 interacting proteins and our data suggested that Raf1 may regulate cell processes through these interactions. These interaction partners were involved in all ten hallmarks of cancer, suggesting that Raf1 is involved in different aspects of carcinogenesis. In addition, we showed that Raf1 interacting proteins were enriched in six signaling pathways and many human diseases. The interaction partners identified in this study may represent oncological candidates for future investigations into Raf1 function. Our findings have provided an overview of Raf1 function from a systems biology perspective.

Proteomic Analysis on Cercariae and Schistosomula in Reference to Potential Proteases Involved in Host Invasion of Schistosoma japonicum Larvae.

Schistosomiasis is a parasitic zoonosis posing great threat to human health. The infection is acquired by larval cercariae penetrating host skin and transforming into juveniles, schistosomula. Proteolytic enzymes secreted from the cercarial acetabular glands are known to aid to the skin penetration, but molecular mechanisms remain largely unclear. To profile the protein composition and identify potential invasive proteases, we developed a new method for simulating cercarial transformation and collecting schistosomula, and for the first time, we compared the proteomes of Schistosoma japonicum cercariae and schistosomula by using in-gel shotgun proteomic analysis. Totally, 1972 proteins were identified in association with ten main biological processes based on Gene Ontology analysis; 46 proteases were detected in cercariae, and among them, 25 proteases disappeared after penetrated. Notably, leishmanolysins and serine and cysteine proteases were found abundant but differentially expressed. Recombinant serine protease SjCE2b and cysteine protease SjCB2 were produced and used for validation of native proteins. Immunofluorescence and Western blotting assays detected SjCE2b and SjCB2 in cercariae but not in schistosomula, suggesting the two enzymes might be consumed upon skin migration. Our data comprehensively chart the proteomic changes during cercarial invasion, revealing the potential proteases involved, providing a platform for the development of molecular anti-infection strategy.