Detection And Analysis Of CXCR4, C-FOX, FYN,PTPN11 Gene Expression In EBV-Transformed Lymphoblasts

[Objective]The early research have been screened four key mole- cules that Epstein-Barr virus-induced lympho- cyte transformation, namely CXCR4, c-FOS, FYN, PTPN11. We used Fluorescence quantitative PCR and Western blot technology to detect the gene expression of CXCR4, c-FOS, FYN, PTPN11 in normal lymphocytes and EBV-transformed lymphoblasts in vitro. This experiment can verify the early gene chip results and provide the experimental basis for studying the molecular mechanism of the EBV induces lymphoma. [Methods]Collected venous blood of 7 healthy blood donation and isolated lympho- cytes, Build a immortalized lymphoblasts cell model use EBV induced lymphocyte transformation in vitro, extracted the cellular proteins and RNA of normal lymphocyte and EBV transf- ormation lymphoblasts, used Western blot and fluorescence quantitative PCR method, detected CXCR4, c-FOS, FYN, PTPN11 gene expression in 7 cases of EBV transformation lymphoblasts in vitro and matched healthy adults. [Result]Lymphocyte immortalized after the EBV transformation, the experiment successful established EBV transformed lymphatic cell model, and isolated normal lymph cells from 7 cases of healthy adults peripheral blood, respectively extracted protein and m RNA, and examinated the CXCR4, c-FOS, FYN, PTPN11 gene expression in 7 cases of normal lymphoid cells and EBV transformated lympho- cytes. [Conclusion]CXCR4 gene expressed in both normal lymphocytes and transformed cells, compared with normal lymph- ocytes, the m RNA expression in transformed cells was down- regulated 2.718 times, the difference is statistically signif- icant(P<0.01); Western blot results showed that, compared with normal lymphocytes, CXCR4 protein expression was down- regulated 1.894 times, the difference was statistically significant(P<0.01).c-FOS gene expressed in both normal lymphocytes and transformed cells, compared with normal lymphocytes, the m RNA expression in transformed cells was down-regulated 2.641 times, the difference is tatistically significant(P<0.01); Western blot results showed that, compared with normal lymphocytes, c-FOS protein expression was down-regulated 2.323 times, the difference was statistically significant(P<0.01). FYN gene expressed in both normal lymphocytes and transformed cells, compared with normal lymphocytes, the m RNA expression in transformed cells was down-regulated 2.451 times, the difference is statistically significant(P<0.01); Western blot results showed that, compared with normal lymphocytes, FYN protein expression was down-regulated 2.019 times, the difference was statistically significant(P<0.01).PTPN11 gene expressed in both normal lymphocytes and transformed cells, compared with normal lymphocytes, the m RNA expression in transformed cells was up-regulated 2.057 times, the difference is statistically significant(P<0.01); Western blot results showed that, compared with normal lymphocytes, PTPN11 protein expression was up-regulated 2.127 times, the difference was statistically significant(P<0.01). [Conclusion]1. The levels of CXCR4 m RNA transcription and protein expression in transformed cells were down-regulated than in normal lymphocytes.2. The levels of c-FOS m RNA transcription and protein expression in transformed cells were down-regulated than in normal lymphocytes. 3. The levels of FYN m RNA transcription and protein expression in transformed cells were down-regulated than in normal lymphocytes.4. The levels of PTPN11 m RNA transcription and protein expression in transformed cells were up-regulated than in normal lymphocytes.