【摘要】 白血病抑制因子(LIF)为一种多功能细胞因子,在生物发育及维持正常生理功能中发挥着重要的作用. LIF在所有的哺乳动物中都具有高度的保守性,某些氨基酸在不同物种中保持不变. LIF中29位氨基酸在所有的哺乳动物中都是Q. 为了研究Q对LIF功能的重要性,利用随机PCR的方法将29位氨基酸突变为R,并将突变后的LIF克隆到真核表达载体pcDNA6,在哺乳动物细胞中成功表达. 同时以克隆到的野生型LIF作为对照. 将分泌到细胞培养基中的野生型和突变LIF因子收集,通过EMSA实验以及荧光素酶报告系统检测,发现野生型LIF因子具有激活STAT3信号通路的生物学活性. 同时,3H-TdR掺入实验结果表明,野生型LIF因子能显著抑制鼠骨髓白血病细胞系M1的增殖。而29位氨基酸突变的LIF因子则完全丧失了以上功能,但所发现的突变没有显性负的作用. 结果充分说明,LIF中高度保守的29位氨基酸对LIF功能的维持具有重要的作用.更多还原

【Abstract】 Leukemia inhibitory factor (LIF) plays important roles in varieties of biological processes. This factor is highly conserved in mammalian animals and only one heterozygous LIF mutation was reported to cause the infertility of women. A LIF mutation was generated and the evidences were provided that the mutation of mature LIF at the 29th amino acid totally abolished its functions, including stimulation of STAT activation assayed by Luciferase reporter gene expression and EMSA experiments. In addition, the mutated LIF failed to inhibit the proliferation of M1 cells. The data indicated that the mutation of LIF did not have a dominant negative effect but lost the biological functions, suggesting that the 29th amino acid is critical for maintaining the activities of LIF.更多还原