HSF4b突变对人晶状体细胞影响的蛋白质组学研究

Objective To study the proteome changes of HLECs,SRA01/04,which were transfected with mutated HSF4b(T to C transition at nucleotide 348).Methods Human lens cells(HLECs),SRA01/04, were cultured in vitro. The plasmid,pcDNA3.1-HSF4b was mutated at certain site. Plasmid was amplified on a large scale. HLECs were transfected with plasmids using lipofection（pcDNA3.1-HSF4b，mutant pcDNA3.1-HSF4b, empty plasmid vector pcDNA3.1）.The whole cell proteins of four groups （three groups were transfected with different plasmid and one group without transfection）were extracted and were labeled with iTRAQ after digestion. The proteome changes were studied using liquid chromatography and mass spectrometry. Results One hundred and four proteins were identified on the whole. Comparing with other three groups, the cells transfected with mutant pcDNA3.1-HSF4b had 20 proteins up-regulated and 14 proteins down-regulated. Twelve proteins in the group transfected with mutant pcDNA3.1-HSF4b and 14 proteins in the group transfected with pcDNA3.1-HSF4b were up- or down-regulated compared with transfected with empty vector pcDNA3.1,among which Vinculin was up-regulated and Protein S100-A13 was down-regulated. Conclusion Some proteins were up- or down-regulated after HLECs were transfected with mutant pcDNA3.1-HSF4b.These changed proteins were related with heat shock response, cell differentiation, protein biosynthesis and DNA binding.