Integrated Glycoproteomics Demonstrates Fucosylated Serum Paraoxonase 1 Alterations in Small Cell Lung Cancer

Small cell lung cancer (SCLC) is an aggressive type of lung cancer, and the detection of SCLCs at an early stage is necessary for successful therapy and for improving cancer survival rates. Fucosylation is one of the most common glycosylation-based modifications. Increased levels of fucosylation have been reported in a number of pathological conditions, including cancers. In this study, we aimed to identify and validate the aberrant and selective fucosylated glycoproteins in the sera of patients with SCLC. Fucosylated glycoproteins were enriched by the Aleuria aurantia lectin column after serum albumin and IgG depletion. In a narrowed down and comparative data analysis of both label-free proteomics and isobaric peptide-tagging chemistry iTRAQ approaches, the fucosylated glycoproteins were identified as up- or down-regulated in the sera of limited disease and extensive disease stage patients with SCLC. Verification was performed by multiple reaction monitoring-mass spectrometry to select reliable markers. Four fucosylated proteins, APCS, C9, SERPINA4, and PON1, were selected and subsequently validated by hybrid A. aurantia lectin ELISA (HLE) and Western blotting. Compared with Western blotting, the HLE analysis of these four proteins produced more optimal diagnostic values for SCLC. The PON1 protein levels were significantly reduced in the sera of patients with SCLC, whereas the fucosylation levels of PON1 were significantly increased. Fucosylated PON1 exhibited an area under curve of 0.91 for the extensive disease stage by HLE, whereas the PON1 protein levels produced an area under curve of 0.82 by Western blot. The glycan structural analysis of PON1 by MS/MS identified a biantennary fucosylated glycan modification consisting of a core + 2HexNAc + 1Fuc at increased levels in the sera of patients with SCLC. In addition, the PON1 levels were decreased in the sera of the Lewis lung carcinoma lung cancer mouse model we data fucosylated protein as PON1, and fucosylation levels and as diagnostic and for SCLC. Small cell lung cancer (SCLC) is an aggressive type of lung cancer, and the detection of SCLCs at an early stage is necessary for successful therapy and for improving cancer survival rates. Fucosylation is one of the most common glycosylation-based modifications. Increased levels of fucosylation have been reported in a number of pathological conditions, including cancers. In this study, we aimed to identify and validate the aberrant and selective fucosylated glycoproteins in the sera of patients with SCLC. Fucosylated glycoproteins were enriched by the Aleuria aurantia lectin column after serum albumin and IgG depletion. In a narrowed down and comparative data analysis of both label-free proteomics and isobaric peptide-tagging chemistry iTRAQ approaches, the fucosylated glycoproteins were identified as up- or down-regulated in the sera of limited disease and extensive disease stage patients with SCLC. Verification was performed by multiple reaction monitoring-mass spectrometry to select reliable markers. Four fucosylated proteins, APCS, C9, SERPINA4, and PON1, were selected and subsequently validated by hybrid A. aurantia lectin ELISA (HLE) and Western blotting. Compared with Western blotting, the HLE analysis of these four proteins produced more optimal diagnostic values for SCLC. The PON1 protein levels were significantly reduced in the sera of patients with SCLC, whereas the fucosylation levels of PON1 were significantly increased. Fucosylated PON1 exhibited an area under curve of 0.91 for the extensive disease stage by HLE, whereas the PON1 protein levels produced an area under curve of 0.82 by Western blot. The glycan structural analysis of PON1 by MS/MS identified a biantennary fucosylated glycan modification consisting of a core + 2HexNAc + 1Fuc at increased levels in the sera of patients with SCLC. In addition, the PON1 levels were decreased in the sera of the Lewis lung carcinoma lung cancer mouse model we data fucosylated protein as PON1, and fucosylation levels and as diagnostic and for SCLC. cancer is the most common of cancer with one of cancer four as cell cell and cell carcinoma of cell lung Small cell lung cancer (SCLC) cell lung disease disease aurantia under lung for and reaction cell lung disease disease aurantia under lung for and reaction is the by of cell lung pathological and of cell lung is a lung cancer for of lung cancer cancer is an aggressive early and a including and the survival for and for cell lung we is by the of of for patients with lung diagnostic and for SCLC, as and in of early the of diagnostic to identify is for the of or to more and of for patients with lung The of serum is a for the and and early detection of cancer, and a of is to and cell lung cancer limited disease stage extensive disease stage Aleuria aurantia lectin area under curve Lewis lung carcinoma isobaric for and multiple reaction serum serum hybrid ELISA Western curve cell lung cancer limited disease stage extensive disease stage Aleuria aurantia lectin area under curve Lewis lung carcinoma isobaric for and multiple reaction serum serum hybrid ELISA Western curve is one of the of proteins for of serum proteins, including and proteins, to the of protein as analysis of the glycan of proteins of the of cell in cancer and in and A. number and of to cell and of by and In have been as of cancer in glycoproteins in and A. of glycoproteins in and A. of proteins produced by and for the and of and cancer The detection of and a to diagnostic and the glycoproteins been the and and a and in of to in including and glycoproteins as of is a of the of glycan and is with cancer and A. number and of to cell and The fucosylation of been reported in as cancer, cancer, cancer, cancer, and cancer of fucosylation in cancer The of fucosylation in cancer is with cancer modification of and cancer cell with Fucosylation and and fucosylated have been as in the sera of patients with and of and The fucosylated of more diagnostic A. 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values were to the performed curve analysis for a of serum the the and the and in the serum levels were significantly increased in the with the The for the and was and this the and was The serum levels were significantly in the and with in the The for the and was and the the and was The serum protein levels of as was as a down-regulated protein in the by analysis exhibited the the patients with and the The the and data to in the of the in to detection The serum levels of PON1 were significantly in the and with in the The for PON1 the and was and the the and was the levels of the fucosylated of the APCS, C9, SERPINA4, and PON1 proteins in the serum the fucosylated glycan levels the four selected proteins, we a fucosylated detection The HLE a to the proteins and a with to the fucosylated the The fucosylated protein levels were by HLE in the sera of and and patients The levels of fucosylated were increased in the sera of the patients with SCLC. were the and the and and the and The for the and was whereas the and was The fucosylation levels were significantly in the with the The for the and was whereas the and was The of fucosylated was decreased in the whereas the protein was the were the and and the and the and The for the and was whereas the and was the fucosylated PON1 levels were significantly increased in the with the and was the and The for PON1 the and was and the and was the of fucosylated PON1 levels was of the PON1 protein levels by the fucosylated PON1 levels or more as a of SCLC. 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A. is in cancer and cell In this study, fucosylated glycoproteins enriched by the column were by of comparative label-free and iTRAQ the four proteins, APCS, C9, SERPINA4, and PON1, were selected for the analysis of protein levels and of fucosylation in and serum The PON1 protein levels were and fucosylation levels were significantly increased in the glycan analysis the fucosylated glycan at is increased in the lung cancer mouse model decreased serum levels of PON1 data the of PON1 fucosylation as an diagnostic for SCLC. performed multiple proteomics to and to more reliable for The of the label-free of proteins and the iTRAQ more we of for the of The of the four proteins were with the Four proteins were selected as the were in a cancer The glycan of the proteins were for these four proteins, the levels of and were significantly whereas PON1 was significantly decreased in the the values of were increased in the and with the was significantly the to both and The and to in the albumin and IgG and fucosylated protein for the analysis were with sera for the by HLE, the fucosylation levels of and were significantly increased as the protein levels increased. the fucosylation levels of PON1 were significantly whereas the PON1 protein was The fucosylation levels of were significantly decreased in the in the of SERPINA4, the fucosylated by HLE a more for the protein exhibited the and in protein levels of SERPINA4, APCS, and PON1 of early stage patients the and the fucosylation levels in these exhibited and PON1, the HLE analysis more and by HLE an for the of patients with In this study, we for the the fucosylated glycan of glycoproteins in the sera of patients with SCLC. glycan analysis fucosylated glycan were increased in and In the glycan at and the glycan at were in the in the these were increased disease to data these as serum for SCLC. protein is with in and SERPINA4, a and was to and in Lewis lung carcinoma mouse In study, the of and the survival of the Lewis lung carcinoma of lung by of The of was to and in a carcinoma mouse model therapy and to carcinoma in of and and in a cancer mouse model protein of carcinoma by and The decreased levels of have been reported in carcinoma with as by with proteomics analysis protein at data a of protein levels in and a more of the fucosylated a for the of the and of SCLC. APCS, a of the been to in and as a protein is with to by serum in in by therapy of of in patients with cancer have been of protein of cancer patients in serum to and after was significantly decreased after of in serum protein in cancer an Increased levels have been reported in the of a model and in serum lung cancer as by proteomics analysis of in by and to identify disease serum proteins in lung cancer of Increased levels reported in of as protein to identify of analysis of in and of protein and in of with were in the levels of in the sera of cancer patients levels in patients with the levels of protein were to decreased in with The of of the The increased levels in by the of and cell is the is a of the a in the by in the of reported fucosylated levels a for lung cancer A. Fucosylated to identify a with cell lung cancer In this study, we both protein levels and fucosylated levels as PON1 to and to the of as and and for of in PON1 is of an in the and been to as of serum and in patients with and in patients with lung A. and in patients with lung cancer in a levels in and levels in cancers. the these were performed PON1 and the of serum The PON1 levels were decreased in these with the a and in early carcinoma reported the increased fucosylation and of serum PON1 as glycan for the early of carcinoma in In study, was to PON1 was the sera of the carcinoma In this study, we the fucosylated PON1 levels by the lectin by HLE, was to the levels of PON1 fucosylation in a number of at a The fucosylated PON1 levels were significantly increased in the sera of the patients with SCLC, whereas the PON1 protein was significantly In addition, we the fucosylated glycan of PON1 for the by the fucosylated glycan at of PON1 was increased in the with data the PON1 the have of fucosylated number of PON1 The serum levels of PON1 protein by the data have PON1 is in lung cancer the PON1 protein levels in the and in mouse model a of PON1 serum with the in the PON1 levels in the were significantly the and is lung cancer the of PON1 in the of PON1 proteins the to the the lung cancer to the of fucosylated PON1 or fucosylated PON1 proteins whereas fucosylated PON1 proteins to reduced in the fucosylated PON1 proteins in the of patients with SCLC. is to the the fucosylated PON1 levels whereas the PON1 levels decreased in lung cancers. more fucosylated PON1 fucosylated PON1 protein more to fucosylated PON1 to the serum proteins and cancer, were the the by and A. A. is in cancer and cell A. of and PON1 is a to cell and to lung cancer PON1 the for to cell in the reduced serum levels of PON1 we in this these to by In fucosylated fucosylated glycan and the glycan of PON1 as for the of SCLC. data to to these a of these with significantly the and of for the of SCLC. with