人参皂苷Rg1诱导白血病K562细胞老化的研究  

Study on the aging of leukemia K562 cells induced by ginsenoside Rg1

作　　者：韩艳军 周玥 HAN Yan-jun;ZHOU Yue(Pre-Clinical College,Dali University,Dali 671000,Yunnan,China)

机构地区：[1]大理大学基础医学院

出　　处：《医学研究生学报》2020年第1期32-37,共6页Journal of Medical Postgraduates

基　　金：国家自然科学基金（81660731,81860038）

摘　　要：目的目前人参皂苷Rg1主要集中于对实体瘤和急性白血病的研究,涉及慢性白血病的报道较少。文中旨在探讨共济失调毛细血管扩张突变基因Rad3相关蛋白(ATR)-检查点激酶1(Chk1)通路在人参皂苷Rg1诱导白血病K562细胞老化中的作用。方法采用不同浓度的人参皂苷Rg1处理K562细胞,分为对照组(加入50μL PBS培养液)、5μmol/L人参皂苷组、10μmol/L人参皂苷组、20μmol/L人参皂苷组、40μmol/L人参皂苷组、80μmol/L人参皂苷组。运用CCK-8法、集落形成实验和流式细胞术检测细胞周期确定人参皂苷Rg1对K562细胞老化的影响;SA-β-Gal染色法和Wright’s染色法观察K562细胞衰老形态学变化;实时荧光定量PCR和Western blot检测细胞老化调控因子ATR和Chk1表达的改变。结果20μmol/L人参皂苷组K562细胞集落形成率较其他组明显降低(P<0.05)。CCK-8检测结果显示,人参皂苷Rg1各组分别诱导K562细胞24、48、72 h,K562细胞增殖能力较对照组升高(P<0.05);当20μmol/L人参皂苷组48 h时K562细胞增殖抑制率最高(P<0.05)。20μmol/L人参皂苷组作用48 h后,K562细胞SA-β-Gal染色阳性细胞率[(95.833±1.528)%]显著高于对照组[(3.083±0.764)%],阻滞在G0/G1期的细胞较对照组明显增高,进入S期和G2/M期的细胞较对照组明显减少(P<0.05);且ATR和Chk1 mRNA表达水平[(0.0117±0.0038)%、(0.0120±0.0021)%]较对照组[(0.0027±0.0006)%、(0.0058±0.0019)%]明显增高(P<0.05);ATR和Chk1蛋白相对表达水平[(19.370±0.994)%、(43.520±1.236)%]较对照组[(17.080±1.274)%、(39.100±0.969)%]明显上调(P<0.05)。结论人参皂苷Rg1可通过调控ATR-Chk1通路诱导K562细胞老化,可为临床白血病治疗提供新靶点。Objective At present,the main studies of ginsenoside Rg1 are almost on the field of solid tumors and acute leukemias,and few on chronic leukemias.We aims to figure out the role of ATR-Chk1 pathway on cell aging in ginsenoside Rg1-treated leukemia K562 cells.Methods K562 cells were treated with ginsenoside Rg1 at different concentrations and divided into a control group(with 50μL PBS culture solution)and 5μmol/L ginsenoside group,10μmol/L ginsenoside group,20μmol/L ginsenoside group,40μmol/L ginsenoside group,80μmol/L ginsenoside group.CCK-8 assay,colony formation assay and flow cytometry for cell cycle detection were used to determine the effect of ginsenoside Rg1 on the aging of K562 cells.SA-β-Gal staining and Wright’s staining were used to observe the morphological changes of K562 cells’aging.Real-time quantitative PCR and Western blot were used to detect the changes of ATR and Chk1 expression.Results The colony formation rate of K562 cells in the 20μmol/L ginsenoside group was significantly lower than that in the other groups(P<0.05).CCK-8 test results showed that K562 cell proliferation of ginsenoside Rg1 induced groups was higher than that of the control group at 24,48,and 72 hours(P<0.05).K562 cell proliferation inhibition rate was the highest in 20μmol/L ginsenoside group for 48 hours treatment(P<0.05).The rate of SA-β-Gal positive cells[(95.833±1.528)%]in 20μmol/L ginsenoside-treated K562 cells for 48 h was significantly higher than that of the control group[(3.083±0.764)%].Cells blocked in G0/G1 phase and entered S and G2/M phases were significantly higher and lower than those in the control group,respectively(P<0.05).The ATR and Chk1 mRNA expression levels[(0.0117±0.0038)%,(0.0120±0.0021)%]were significantly higher than that of the control group([0.0027±0.0006)%,(0.0058±0.0019)%)(P<0.05).ATR and Chk1 relative protein expression levels[(19.370±0.994)%,(43.520±1.236)%]were significantly increased compared with that of the control group[(17.080±1.274)%,(39.100±0.969)%)](P<0.05).Co

关 键 词：人参皂苷RG1 K562细胞 老化 共济失调毛细血管扩张突变基因Rad3相关蛋白-检查点激酶1通路 

分 类 号：R552[医药卫生—血液循环系统疾病]