Study On Influence On The Expressive Spectrum Of Three Kinds Of Glycosyltransferase Gene Family In Different Leukemia Cells By The Differentiation Of All-transretinoic Acid

Objective: To observe the expression of polypeptide : N-acetylgalactosaminyl transferase Ts(ppGalNAcTs) and UDP-N-acetylglucosamine:beta-galactose beta 1,3-N-acetylglucosaminyltransferase(β3GnT1, β3GnT3, β3GnT4, β3GnT5), O-Linked GlcNAc transferase (OGnT) in different leukemia cells including HL60 cell line and SHI-1 cell line and to study the variation of acetylglucosaminy transferase expression after ATRA 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 ATRA on ppGalNAcTs and β3GnT1, β3GnT3, β3GnT5, OGnT.Methods: 1. Use RT-PCR and real time RT-PCR to detect (1) the mRNA expression of ppGalNAcTs and β3GnT1, β3GnT3, β3GnT4, β3GnT5, OGnT in HL60 cell line and SHI-1 cell line;(2)the expression discrepancy of ppGalNAcTs and β3GnT1 , ppGalNAcT2, ppGalNAcT4, β3GnT5, OGnT in HL60 cell line and SHI-1 cell line after ATRA added to the culture medium.2 . Use the assay of radioactive compounds ~3H-thymidine incorporation method to study the livability of leukemia cells induced by ATRA.3.To study the cell circle of leukemia cells induced by ATRA with the flow cytometer.4. To study the change of cell shape of leukemia cells induced by ATRA with the LaserConfocal Scanning Microscope(LCSM).Results: 1. The expressions of glycosyltransferase were different in two leukemia celllines.2. After ATRA added to the culture medium, the mRNA expression of p3GnTKppGalNAcT2 in HL60 cell line greatly increased. The cell cycle was arrested in G2/Mphase ; No cell death was detected. The expression of glycosyltransferase in SHI-1 cellline has no obvious change than in HL60 cell line.Conclusion: This study shows that the expression of glycosyltransferase in differentleukemia cells is different. This result suggests that different glycosyltransferases havedifferent particular substances. Leukemia cell differentiation probably has contact withP3GnTl > ppGalNAcT2.