Targeted Screening Of Rab14-interacted Proteins In HBV Related Hepatocellular Carcinoma

Aim:Targeted proteomics was used to identify Rab14-interacted proteins in hepatitis B virus(HBV) related hepatocellular carcinoma (HCC). Bioinformatics methods including gene ontology(GO), functional cluster, signal pathway and protein-protein interaction(PPI) was performed to analyze Rab14-interacted proteins to reveal biological significance and explore possible molecular mechanism of Rab14in HBV related HCC.Methods:1Real-time quantitative polymerase chain reaction and western blot were used to detect the expression of Rab14in HCC tissues.2Targeted proteomics(co-immunoprecipitation coupled with mass spectrometry) was used to identify Rabl4-interacted proteins.3DAVID v6.7, a bioinformatics software, was used to analyze the Rab14-interacted proteins, which include GO analysis, functional cluster analysis and signal pathway analysis. The string9.1, a software to predict PPI, was used to analyze PPI.Results:1When the Rabl4in HCC tissues was compared with those in surrounding non-tumor tissues or the normal liver tissues, its expressional level was up-regulated significantly(P<0.05).2One hundred and thirty-three Rab14-interacted proteins were identified by targeted proteomics in purified HCC cells.3GO analysis showed that one hundred and thirty-three Rab14-interacted proteins were grouped into13functional classes. Functional cluster analysis showed that65Rabl4-interacted proteins had similar structure and function, which involved9functional clusters:molecular chaperone, vesicle-mediated transport, cytoskeleton, GTP binding, protein transport, Ras small GTPase(Rab type), transmembrane transport, ribosome, ion transport.4Biocarta and KEGG pathway analyses showed that fifteen Rab14-interacted proteins were involved in4Biocarta pathways:shuttle for transfer of acetyl groups from mitochondria to the cytosol, cell to cell adhesion signaling, Rab GTPases mark targets in the endocytotic machinery and the role of FYVE-finger proteins in vesicle transport. Fifty three Rab14-interacted proteins were involved in7KEGG pathways: antigen processing and presentation, adherens junction, endocytosis, ribosome, regulation of actin cytoskeleton, pathways in cancer, MAPK signaling pathway. Among them,3proteins were involved in vesicle transport pathway.9proteins were involved in endocytosis pathway and7proteins were involved in MAPK signaling pathway.5PPI analysis showed that Rab14interacted with TFRC and AP1G1through RAB5A. Rab14interacted with DNM2, CLTC, FTL and TFRC through AP1B1. Rab14interacted with CTNNA1, AP1G1, AP1B1, TF, TFRC and VCL through DNM2.Conclusions:1The expressional levels of Rab14in patients with HCC were up-regulated significantly.2For the first time, One hundred and thirty-three Rab14-interacted proteins were identified by targeted proteomics in HCC cells.3One hundred and thirty-three Rab14-interacted proteins were grouped into13functional classes, which involved in9functional clusters:molecular chaperone, vesicle-mediated transport, cytoskeleton, GTP binding, protein transport, Ras small GTPase(Rab type), transmembrane transport, ribosome, ion transport.4Rab14-interacted proteins involved in4Biocarta signaling pathways and7KEGG signaling pathways.5Rab14interacted with TFRC and AP1G1through RAB5A. Rab14interacted with DNM2, CLTC, FTL and TFRC through AP1B1. Rab14interacted with CTNNA1, AP1G1, AP1B1, TF, TFRC and VCL through DNM2. These regulated vesicle transport, endocytosis and cell adhesion, and so on. The proteins may participated in the occurrence and metastasis of HCC.