Primary Study Of The Relationship Between Human β1,3-N-acetylglucosaminyltransferase Gene β3GnT8 (GalT7) And β3GnT2 In Different Leukemia Cells

Object:β1,3-N-acetylglucosaminyltransferase 8 is a new member of glycosyltransferases we gained by electric cloning. We considered it belongs to beta-3 glycosyltransferase family.It is reported thatβ3GnT2 andβ3GnT8(GalT7) can form a complex with enhanced enzymatic activity. This thesis aims to study the expression ofβ3GnT2 andβ3GnT8(GalT7) gene in different leukemia cells, thereby study on the relationship between the two gene in leukemia.Methods: 1.RT-PCR was used to detect the mRNA expression in 8 leukemia cells,24 patients and bone marrow of 10 normal persons. 2. Constructed the RNA interference expression plasmid ofβ3GnT8(GalT7), and obtained stable cell line withβ3GnT8(GalT7) knockdown expression. 3. The expression discrepancy ofβ3GnT2 andβ3GnT8(GalT7) gene between the normal cells and transfected cells were studied by RT-PCR and Western blot method.Results: 1. Different expression level ofβ3GnT8(GalT7) can be found in leukemia cells SHI-1, THP-1, HL-60, K562, NB4, U937 and 18 patients who were leukemia at first diagnosis, but never found in leukemia cell Raji,KG1 and bone marrow of normal persons;β3GnT2 expressed in SHI-1,THP-1,HL-60,K562,NB4,U937,Raji,16 patients and 4 bone marrow of normal persons, never found in KG1 and 6 bone marrow of normal persons. 2. Constructed the RNA interference expression plasmid ofβ3GnT8(GalT7). 3. The result of RT-PCR and Western blot indicated the expression of theβ3GnT8(GalT7) andβ3GnT2 were declined both on the mRNA and protein level as expression ofβ3GnT8(GalT7).Conclusion: Different expression level ofβ3GnT2 andβ3GnT8(GalT7) can be found in different leukemia cells suggested thatβ3GnT2 andβ3GnT8(GalT7) gene have certain relationship influence on the tumor infiltration and transfusion.