【摘要】 以杂交瘤细胞为模型 ,在培养液中加入不同的信号转导抑制剂和囊素 ,用间接ELISA法测定抗体分泌水平 ,用定量RT PCR法检测抗体mRNA的表达水平。结果表明 ,加入Gα 蛋白选择性抑制剂NF 0 2 3、蛋白激酶C抑制剂GF10 92 0 3X、磷脂酰肌醇 3激酶抑制剂heparin、Ca2 + /calmodulin依赖蛋白激酶Ⅱ抑制剂KN 6 2、磷脂酶C抑制剂U 7312 2和蛋白激酶G抑制剂后 ,囊素对杂交瘤细胞抗体分泌速度的影响明显 (P <0 0 1)。而加入蛋白激酶A抑制剂 4 Cyano 3 M thylisoquinoline、酪氨酸蛋白激酶抑制剂 genistein和丝裂原活化的蛋白激酶抑制剂PD 980 5 9后 ,囊素对杂交瘤细胞抗体分泌速度的影响不明显 (P >0 0 5 ) ,这 3种抑制剂对杂交瘤细胞抗体分泌速度的抑制率分别为 74 5 5 % ,91 13%和80 6 9%。加入 4 Cyano 3 M thylisoquinoline、genistein和PD 980 5 9后 ,囊素处理组与对照组γ1mRNA的表达水平差异不显著 (P >0 0 5 ) ,说明这 3种抑制剂阻断了囊素刺激的杂交瘤细胞抗体mRNA的表达。以上结果表明 ,蛋白激酶A、酪氨酸蛋白激酶和丝裂原活化的蛋白激酶参与了囊素在杂交瘤细胞中的信号转导。更多还原

【Abstract】 In order to investigate the influences of signal transduction inhibitors on the antibody production and mRNA expression in hybridoma cells promoted by bursin,hybridoma cells pre-treated by different inhibitors were stimulated by bursin,then antibody levels and mRNA expression levels of which were tested by indirect ELISA and semi-quantitative RT-PCR respectively.The results showed that,after treatment with NF023,GF109203X,heparin,KN-62,U-73122 and inhibitor of protein kinase G,bursin could still greatly promote the production of monoclonal antibody in hybridoma cells(P<0.01),while bursin played no role on the production of monoclonal antibody in hybridoma cells pre-treated by 4-Cyano-3-M-thylisoquinoline,genistein and PD98059 respectively.The inhibiting efficiency of 4- Cyano-3-M- thylisoquinoline,genistein and PD98059 on the production rate of monoclonal antibody in hybridoma cells were 74.55%,91.13% and 80.69% respectively.The expression levels of γ 1 mRNA in hybridoma cells treated with bursin and 4-Cyano-3-M-thylisoquinoline,genistein or PD98059 were 1.845±0.236,1.632±0.476 and 1.728±0.353 respectively,showed no significant difference from that of hybridoma cells stimulated with 4-Cyano-3-M-thylisoquinoline,genistein or PD98059 alone(P>0.05).These results indicated that protein kinase A(PKA),tyrosine kinase and mitogen-activated protein kinase kinase(MAPKK) were involved in the signal transduction of bursin on hybridoma cells.更多还原