Design And Fabrication Of Glycan-binding Protein Gene Microarray

Background:The interactions between glycan chains of cell surface and proteins can promote the cell-cell adhesion and vertebrate embryo growth. This kind of proteins is called glycan-binding protein (GBP). GBPs can mediate cell migration, signaling transduction, regulation between cells, between cells and matrix reaction by identifying glycan chains of cell surface or in body fluid; it also play an important role in the occurrence and development of the tumor. Liver cancer cntain some features, including:can not detected early; easy to transfer; development fast and difficult to treat. GBPs and its ligand glycan chains have significant change in liver cancer cells. Also the genes of GBPs are changed. So it is indispensable to monitor the expression profile of the GBP gene expression for the investigation of other tumor.Method:We use gene chip technology which is a high-flux detecting method to investigate the variety Expression GBP gene between liver cancer and normal liver cells/blood. Select the differential expressed GBP gene. Using the cell lines (HepG2, SMMC-7721, LO2) and the blood samples (liver cancer patient and health people) as the research objects. Using the oligonucleotide arrays which we have designed and prepared (including 135 GBP gene and 25 control gene) to hybridize. Select the variety expressed GBP gene. The result of oligonucleotide arrays is confirmed by Quantitative RT-PCR.Result:With the glycan-binding protein gene chip to detect the cells and the blood samples, we found 24 differential expressed glycan-binding protein gene between LO2 and HepG2:15 high expression and 9 low expression; 15 differential expressed betweenLO2 and SMMC-7721:9 high expression and 6 low expression; 17 differential expressed between the blood samples:9 high expression and 8 low expression. There are 7 common up regulated and 3 common down regulated glycan-binding protein gene in cell lines and blood samples. Using the Quantitative RT-PCR confirm some of the result from oligonucleotide arrays (including ALPP, Galectin-1, MGL and REG4). Conclusion:Using the gene chip and Quantitative RT-PCR technology, we found common variety expressed GBP gene between liver cancer and normal liver Cells/blood. It' meaningful to prevent, diagnose, predict liver cancer, and establish a platform for the research of other tumors.