The Biological Functions Of Pp-GalNac-T In The Difference Of Leukemia

Part1 and Part2 Study on Expressive Spectrum of ppGalNAcTs in Different Leukemia Cells and the Effects of differentiationObjective: To observe the expression of polypeptide : N-acetylgalactosaminyl transferase Ts(ppGalNAcTs) in different leukemia cells including K562 cell line and SHI-1 cell line and to study the variation of acetylglucosaminy transferase expression after 1,25(OH)2 D3 added to the culture medium, to find out the relative function of acetylglucosaminy transferase in the course of leukemias generating and developing, as well as to make out the influence of 1,25(OH)2 D3 on ppGalNAcTs.Methods: 1. Use RT-PCR to detect (1) the mRNA expression of ppGalNAcTs in K562 cell line and SHI-1 cell line;(2)the expression discrepancy of ppGalNAcTs in K562 cell line and SHI-1 cell line after 1,25(OH)2 D3 added to the culture medium. 3.To study the cell circle of leukemia cells induced by 1,25(OH)2 D3 with the flow cytometer. 4. To study the change of cell shape of leukemia cells induced by1,25(OH)2 D3 with the Laser Confocal Scanning Microscope(LCSM). Results:1. The expressions of glycosyltransferase were different in two leukemia cell lines.2. Afte 1,25(OH)2 D3 added to the culture medium, the mRNA expression of ppGalNAcT2、T5 in K562 cell line greatly increased. The cell cycle was arrested in G2/M phase ; No cell death was detected. The expression of glycosyltransferase in SHI-1 cell line has no obvious change than in K562 cell line.Conclusion: This study shows that the expression of glycosyltransferase in different leukemia cells is different. This result suggests that different glycosyltransferases have different particular substances. Leukemia cell differentiation probably has contact with pp-GalNAcT2. T5.Part3 and Part4 Construction of Plus Sense Expression plasmid of pp-GalNAc-T 4 gene and Primary Study on its FunctionObjective: This thesis aims to approach the function of pp-GalNAc-T 4 in the differentiation of leukemia induced by ATRA.Methods:1, Subclone pp-GalNAc-T4 fragment to eukaryotic plasmid and form plus sense plasmids. Transfect the recombinant plasmids separately to NB4. To study the change of cell shape of leukemia cells induced by ATRA with the Laser Confocal Scanning Microscope(LCSM)..Studied the variational characteristics of NBT and PML-RARαexpression on mRNA level with Real Time -PCR.Results:1. Subclone pp-GalNAc-T4 fragment to eukaryotic plasmid and form plus sense plasmids2.Up-regulated expressions of pp-GalNAc-T4,inhibited differentiation of leukemia NB4 induced by ATRA.2. Up-regulated expressions of pp-GalNAc-T4,inhibited differentiation of leukemia NB4 induced by ATRA .PML-RARαmay contribute to this functions. CD44 may contribute to this functions that cannot the degradation of PML-RARαConclusion: Up-regulated expressions of pp-GalNAc-T4,inhibited differentiation of leukemia NB4 induced by ATRA .PML-RARαmay contribute to this functions. CD44 may contribute to this functions that cannot the degradation of PML-RARα.