The Expression Of The Human UDP-Gal: BetaGlcNAc β1,3-N-acetylglucosaminyltransferase 8 Gene In Glioma Cells SHG44 And U251 And Its Protein Distribution Inside The Cells

Objectiveβ1,3-N-acetylglucosaminyltransferase 8 gene is a new member of glycosyltransferases we gained by electric cloning. We considered it belongs to beta-3 glycosyltransferase family. This thesis aims to study the expression ofβ3GnT8 gene in glioma cells SHG44 and U251, thereby study on the location of the protein in cells. Method 1. RT-PCR was used to detect the mRNA expression of GnT8; 2. Western blot was used to detect the protein expression; 3.Use immunocytochemistry identification the distribution of GnT8 in two glioma cells Result 1.The two cells both erpress GnT8,the expression low in SHG44 but higher in U251; 2.The protein expression is the same to mRNA,low in SHG44 and higher in U251; 3 cellular immunofluorescence stain showed that the protein expressed in cytoplasm around nucleolus in two glioma cells.Conclusion The mRNA and protein expression ofβ3GnT8 gene in glioma cell SHG44 were both low, but higher in U251. The protein mainly distributing in cytoplasm around nucleolus.It suggests the protein distributed on the golgi apparatus which has intimate relationship with the protein synthesis and elaboration.