Detection And Analysis Of TOP2A, PBK,HIST2H2BE Gene Expression In EBV-Transformed Lymphoblasts

[Objective]The early researches have screened three molecules which may play a key role in the process of Epstein-Barr virus-induced lymphoma by gene chip examination, respectively TOP2 A,PBK, HIST2H2 BE. The expression of TOP2 A, PBK, and HIST2H2 BE genes was detected by real-time fluorescence quantitative PCR and Western blot technology in both EBV-transformed lymphoblasts and normal lymphocytes in vitro to verify the results of early gene chip and provide the experimental basis for the molecular mechanism of EBV-induced lymphocyte transformation and occurrence of lymphoma.[Method]We collected peripheral venous blood from seven healthy donations and isolated lymphocytes, and then built transformed lymphoblastoid with EBV-induced lymphocyte transformation in vitro.Total RNA and protein were extracted from the EBV-transformed lymphoblasts and the matched normal lymphocytes, and then real-time fluorescence quantitative PCR and Western blot technology were used to detect the expression of TOP2 A, PBK, HIST2H2 BE genes in seven cases of EBV-transformed lymphoblasts and the matched lymphocytes fromhealthy adults.[Result]the experiment successfully established EBV-transformed lymphoblastoid cell lines,and detected the expression of TOP2 A, PBK,HIST2H2 BE genes in seven cases of EBV-transformated lymphoblasts and the matched normal lymphocytes.TOP2 A gene expressed in both normal lymphocytes and transformed cells, compared with normal lymphocytes, the expression of mRNA in transformed cells was up-regulated 1.413 times, the difference is statistically significant(P<0.001); Western blot results showed that, compared with normal lymphocytes, the protein expression of TOP2 A was up-regulated2.760 times, and the difference was also statistically significant(P<0.001).PBK gene expressed in both normal lymphocytes and transformed cells,compared with normal lymphocytes, the expression of mRNA in transformed cells was up-regulated 1.422 times, the difference is tatistically significant(P<0.001); Western blot results showed that,compared with normal lymphocytes, the protein expression of PBK was up-regulated 3.047 times,and the difference was also statistically significant(P<0.01).HIST2H2 BE gene expressed in both normal lymphocytes and transformed cells, compared with normal lymphocytes,the expression of mRNA in transformed cells was up-regulated 1.757 times, the difference is statistically significant(P<0.001); Western blot results showed that, compared with normal lymphocytes,the proteinexpression of HIST2H2 BE was up-regulated 2.003 times, and the difference was also statistically significant(P<0.001).[Conclusion]The levels of TOP2 A, PBK, HIST2H2 BE mRNA transcription and protein expression in transformed cells were up-regulated than those in normal lymphocytes.