The Glycosphingolipidomics View: The Study Of Novel Glycosphingolipids In HCC

PartⅠThe expression of glycosphingolipids in hcc solid tumortissues and paired peritumornal tissues by LTQ-MS spectrum method.Objective: To detect the expression of glycosphingolipids in hcc solid tomortissues and paired solid peritumornal tissues by glycomics method, and identify the theaberrant GSLs structures specific to hcc, and examine the power of the aberrant GSLsfor the hcc diagnosis and therapy. Provide the initial structure basis for the study aboutthe function of aberrant GSLs in hcc.Methods: Total lipids was extracted from hcc tumor tissues and pairedperitumornal tissues by component organic solvent. Netutral GSLs and acid GSLs wereseparated by DEAE-Sephadex A25column chromatography. Modifiy GSLs byacetylation and purified by MgSiO4column chromatography then deacetylation.HPTLC analyze the purified GSLs from tumor tissues and paired peritumornal tissuesrespectively. Modify the GSLs by permethylation and analyze by LTQ-ESI MS. Treatthe MS data by SPCC and draw ROC curve.Results:(1) The MS1pattern of GSLs from hcc tumor solid tissues showed anovel group of GSLs at ranges from m/z1600-m/z2000, when compared with the GSLsfrom paired peritumornal tissues.(2) MS2profile of the aberrant GSLs in hccdemonstrated these GSLs were fucosylated structures. These aberrant GSLs wasidentified to be three groups, m/z1086, m/z1260, m/z1290respectively.(3) Receiveroperating characteristic curve (ROC) analysis of the data about MS1patternrespectively from hcc solid tumor tissues and paired peritumornal tissues by SPSSsoftware showed that the AUC of m/z1086was0.809, the AUC of m/z1260was0.722and the AUC of the m/z1290was0.953.Conclusion: The expression of GSLs in hcc tumor tissues were different from those in paired peritumornal tissues, which was reflected by the aberrant expression of agroup of GSLs ranges from m/z1600-m/z2000in the MS1pattern. All three groups ofthe aberrant GSLs was fucosylated. ROC analysis of three groups of GSLs showed thepower of diagnosis, which revealed the potential value of targets of diagnosis andtherapy for hcc. Part Ⅱthe qualitative analysis of aberrant fucosylated GSLs in hcctumor solid tissues and the structure basic for potential target tohcc.Objective: qualitative analysis of the aberrant fucosylated GSLs by LTQ-ESI MSn,and find out the potential structures for target for hcc diagnosis and therapy.Methods:(1) Multistep MS analysis of aberrant fucosylated GSLs;(2) analyze thestructure combining with the Chemdarw software;(3) Multistep MS analysis ofexpression of GSLs in hcc cell lines;(4) Multistep MS analysis of expression of GSLsin serum from hcc patient;(5) Real-time PCR method measure the expression offucosyltransferase responsible for the fucosylation of these aberrant GSLs in hcc solidtumor tissues and paired peritumornal tissues.Results:(1) The result demonstrated that aberrant fucosylated GSLs containedseven structures, which were Type Ⅰ/Ⅱ H antigen，Lewis x，Lewis y，Lewis a，Lewisb，Globo H respectively. Moreover, fucosylated pentosaccharide at m/z1086mainlycontained Type H antigen, double fucosylated at m/z1260mainly contained Lewis band Lewis y, fucosylated at m/z1290was Globo H;(2) the similar result exhibited inthree types of hcc cell line, revealed that the same fucosylated GSLs were contained inhcc cell lines, such as m/z1290;(3) MS pattern of GSLs from hcc patient serumsuggested that the same results exhibited;(4) Treat our MS data in glycomice views,thess aberrant GSLs contained the same terminal Fucɑ1-2Gal-typology, whose MSndata were similar to the Fucɑ1-2Gal-typology standard;(4) the expression of the P value of FUT2was less than0.01.Conclusion: The aberrant fucosylated GSLs contained various of structure, like Hantigen, Globo H et al., in hcc solid tumor tissues. The result revealed that the aberrantfucosylation was complex in hcc. The feature of sugar chains of GSLs led us tohypothesize that taking the common typologies of various tumor-associated GSLantigens as target for tumor therapy. In my study, we found that the aberrant fucosylatedGSLs contained terminal Fuc ɑ1-2Gal-typology, and the AUC value of Fuc ɑ1-2Gal-typology was0.864, which showed the potential value for hcc diagnosis and therapy.The similar result exhibited in the patient serum and hcc cell lines.