Generation of H/Rª²Sª²like murine cells by LMP1 overexpression in A20

    LI Xianª²Mao,ZHU Meiª²Gang£¬SHEN Liª²Jia£¬LIU Tengª²Fei£¬ZHANG Sanª²Quan£¬ZHANG Jinª²Hua£¬ZHANG Yan£¬ZHAO Tong

    Department of Pathology,School of Basic Medicine,Southern Medical University,Guangzhou 510515,China

    ¡¾Abstract¡¿ AIM: To establish an H/Rª²Sª²like murine cell line A20RS by transfecting CMV promoter driven LMP1 into a murine neoplastic B cell line,in which the level of endogenous LMP1 expression is almost negligible.METHODS: The fragment including all the exons of LMP1 was amplified by RTª²PCR and was recombined to the downstream of CMV promoter in the vector PLXSN.PLXSNª²LMP1 and PLXSN were introduced respectively into A20 cell line using lipofectamine method.RESULTS: The sequence of the RTª²PCR product was totally consistent with the data of Gen Bank by DNA sequencing analysis.The LMP1 cDNA fragment was cloned in the vector PLXSN in the right direction and the open reading fragment of LMP1 was maintained. In this cell line A20RS transfected with PLXSNª²LMP1,Western analysis demonstrated the overexpression of LMP1.The morphological characteristics of the cells were almost identical to those of human H/Rª²S cells in lymph nodes of Hodgkin¬ðs disease patients.CONCLUSION: H/Rª²Sª²like murine cell models are generated,which provides a solid basis for the establishment of animal models. ......