STAT3反义表达载体的构建、稳定表达及对M1白血病细胞表型的影响

目的 探讨JAK／STATs通路在IL－6信号转导中的功能。方法 将含起始密码子在内的约1．2kbSTAT3cDNA反向插入到真核细胞表达载体pcDNA3中，采用电穿孔法重组质粒导入到M1小鼠急性髓系白血病细胞中，进行G418筛选。结果 G418筛选出抗性克隆后，基因组DNAPCR扩增表明STAT3反义基因片段已导入M1细胞中并在基因组中整合。STAT3反义RNA不影响M1细胞的生长动力学特征，但可显著拮抗IL－6诱导的STAT3激活，并拮抗IL－6对M1细胞的生长抑制效应。结论 STAT3反义RNA能够在M1细胞中稳定表达，成为进一步研究JAK／STATs途径功能的新模型。

Construction, stable expression of STAT3 antisense expression vector and its effect on the phenotype of M1 leukemia cells

Objective To investigate the biological roles of JAK STATs pathway in the signal transduction induced by IL 6. Methods STAT3 antisense expression vector was constructed. The recombinant vector (pST/Fan) or the vector alone (pcDNA3) was transfected into M1 leukemia cells with electroporation technique. Following selection with neomycin, resistant colonies were obtained. The activation of STAT3 induced by IL 6 was analyzed. Results STAT3 antisense RNA antagonized the IL 6 induced activation of STAT3 efficiently and reduced the inhibitory effect of IL 6 on the growth of M1 cells. However, STAT3 antisense RNA itself did not change the kinetic characteristics of M1 cells. Conclusion The STAT3 antisense RNA modified M1 cells may be new model for studying the function of JAK STATs pathway.