[关键词]
[摘要]
目的:探讨黏蛋白1(mucin 1,MUC1)多肽对人T淋巴瘤Jurkat 细胞生长的抑制作用及其机制。方法:将MUC1多肽与Jurkat细胞共同培养,锥虫蓝染色法观察MUC1多肽对该细胞生长的影响;流式细胞术检测Jurkat细胞生长周期及其细胞表面MUC1的表达;Annexin V/PI双标记法检测Jurkat细胞的凋亡;应用抗体封闭实验检测MUC1多肽在Jurkat细胞表面作用的位点。结果: 10、20、40 μg/ml MUC1多肽作用使Jurkat细胞生长抑制分别达(32±4)%、(37±2)%、(46±5)%,未见凋亡细胞。流式细胞术检测结果表明MUC1多肽可诱导Jurkat细胞周期阻滞在G0/G1期,在Jurkat细胞表面有MUC1蛋白的表达。采用5 μg/ml和25 μg/ml 抗MUC1抗体封闭Jurkat细胞表面MUC1表位后,MUC1多肽对细胞生长的抑制效应几乎全部消失。结论: MUC1多肽能明显抑制Jurkat细胞生长,其机制与该多肽通过和Jurkat细胞表面MUC1蛋白相互作用导致细胞周期阻滞在G0/G1期有关。
[Key word]
[Abstract]
Objective: To investigate the inhibitory effect of mucin 1 (MUC1) peptide on the proliferation of T lymphoma Jurkat cells and the related mechanism. Methods: Jurkat cells were cocultured with MUC1 peptide and effects of MUC1 peptide on Jurkat cell growth was examined by Trypan Blue staining. Cell cycle of Jurkat cells and membrane MUC1 protein expression were determined by flow cytometry. The apoptosis of Jurkat cells was examined by Annexin V/PI double staining. To further analyze whether soluble MUC1 peptide interacts with membrane MUC1 protein, MUC1 protein binding sites on the surface of Jurkat cells was blocked by an antiMUC1 polyclonal antibody. Results: Proliferation of Jurkat cells was inhibited by soluble MUC1 peptide in a dosedependent manner (MUC1 at 10, 20, and 40 μg/ml resulted in inhibitory rates of (32±4)%, (37±2)%, and(46±5)%, respectively ). No apoptosis was observed. Soluble MUC1 peptide induced G0/G1 phase cell cycle arrest of Jurkat cells and membrane MUC1 protein was expressed on the surface of Jurkat cells. The inhibition of Jurkat cells by soluble MUC1 peptide was almost totally reversed by antiMUC1 polyclonal antibodies at the concentrations of 5 μg/ml to 25 μg/ml. Conclusion: It is suggested that soluble MUC1 peptide can inhibit proliferation of Jurkat cells by interacting with membrane MUC1 protein on Jurkat cells and inducing G0/G1 phase cell cycle arrest.
[中图分类号]
[基金项目]
吉林省科技发展计划项目(No.20080931)