摘要
目的探讨人参皂苷Rg1(Rg1)诱导白血病K562细胞衰老及其机制。方法 MTT比色法筛选细胞增殖抑制的最佳浓度及时间,并以此浓度和作用时间进行细胞衰老的相关机理研究。流式细胞术检测细胞增殖周期;SA-β-Gal染色检测阳性细胞;Colony-Assay检测集落形成能力;Southern blotting检测端粒长度;Western blot检测蛋白表达;透射电镜观察细胞超微形态。结果 Rg1在体外能明显抑制K562细胞增殖,使K562细胞阻滞于G2/M期;并显著提高SA-β-Gal染色阳性细胞百分率(P<0.05);降低细胞形成集落的能力;衰老相关蛋白P53、P21、P16及RB表达上调(P<0.05);端粒长度缩短(P<0.05);胞体增大,溶酶体体积增大、数目增多,线粒体体积增大,异染色质凝集等衰老形态学变化。结论人身皂苷Rg1能诱导K562细胞衰老。
Objective To observe the senescence effects and mechanisms of leukemia K562 cell line induced by ginsenoside Rg1（Rg1）.Methods The related mechanism was examined via inducing k562 cells senescence by optimal drug concentration and interaction time detected by MTT colorimetric test.Effect of Rg1 on cell cycle was analyzed using flow cytometry.The positive cells were detected by SA-β-Gal staining.The colony-formaction was detected by Colony-Assay.The telomere lengths were detected by Southern blotting.The expression of proteins was detected by Western blotting and the ultrastructure changes were observed by transmission electron microscopy.Results Rg1 significantly inhibited the proliferation of K562 cells in vitro and arrested the cells in G2/M phase.The percentage of positive cells stained by SA-β-Gal was dramatically increased（P0.05）and the colony-formed ability in experimental group has been weakened（P0.05）.Senescence-related proteins P53、P21、P16、RB in experimental group were up-regulated（P0.05）.Telomere length became shorter.The observation of ultrastructure showed that cell volume increased,heterochromatin condensation and fragmentation,mitochondrial volume increased,lysosomes increased in size and number.Conclusions Rg1 may induce the senescence of leukemia cell line K562 and P53-P21-Rb,P16-Rb cell signaling pathway play a significant role in this process.
出处
《基础医学与临床》
CSCD
北大核心
2012年第5期481-486,共6页
Basic and Clinical Medicine
基金
国家自然科学基金(30973818)
重庆市科委自然科学基金(CSTC2009BA5038)
