摘要
目的探讨急性淋巴细胞白血病(ALL)患者骨髓基质细胞(BMSCs)对Jurkat细胞的黏附作用及小白菊内酯(PTL)对此作用的干预机制。方法采集2013年10月—2014年3月于桂林医学院附属医院经FAB分型及MICM分型确诊为ALL并经治疗后缓解的6例患者髂后上棘骨髓液,体外分离培养BMSCs。Jurkat细胞复苏后传代培养,取生长状态良好的细胞用于实验,建立BMSCs与Jurkat细胞共培养模型。设置Jurkat细胞组、BMSCs组、BMSCs+Jurkat细胞组、BMSCs+Jurkat细胞+PTL 4μmol/L组、BMSCs+Jurkat细胞+PTL 8μmol/L组、BMSCs+Jurkat细胞+PTL 12μmol/L组、BMSCs+Jurkat细胞+PTL 16μmol/L组。黏附实验检测共培养24 h和48 h后各组黏附率,ELISA测定各组可溶性血管细胞黏附分子-1(s VCAM-1)表达水平,RT-PCR法检测BMSCs VCAM-1 mRNA的相对表达量。结果各组培养24 h和48 h后,黏附率与PTL浓度呈负相关(r=-0.95、-0.91,P<0.05)。各组s VCAM-1表达水平比较,差异有统计学意义(P<0.05),BMSCs+Jurkat细胞组、BMSCs+Jurkat细胞+PTL 4μmol/L组、BMSCs+Jurkat细胞+PTL 8μmol/L组和BMSCs+Jurkat细胞+PTL 12μmol/L组s VCAM-1表达水平高于BMSCs组(P<0.05),各加入PTL组s VCAM-1表达水平低于BMSCs+Jurkat细胞组(P<0.05),各加入PTL组s VCAM-1表达水平与PTL浓度呈负相关(r=-0.994,P=0.001)。各组BMSCs VCAM-1 mRNA表达水平比较,差异有统计学意义(P<0.05),BMSCs组与BMSCs+Jurkat细胞+PTL 16μmol/L组BMSCs VCAM-1 mRNA表达水平比较,差异无统计学意义(P>0.05)。各加入PTL组BMSCs VCAM-1 mRNA表达水平与PTL浓度呈负相关(r=-0.994,P=0.001)。结论 PTL可以通过降低VCAM-1的表达抑制BMSCs对Jurkat细胞的黏附,减弱骨髓微环境对白血病细胞的保护作用。
Objective To investigate the adhesion of bone marrow stromal cells（ BMSCs） to Jurkat cells and the intervention of parthenolide（ PTL） on it in patients with acute lymphoblastic leukemia（ ALL）. Methods From October 2013 to March 2014,in the Affiliated Hospital of Guilin Medical College,a co- culture model of BMSCs and Jurkat cells were builted from the posterior superior iliac spine marrow fluid of 6 ALL patients. Jurkat cells group,BMSCs group,BMSCs ＋ Jurkat cells group,BMSCs ＋ Jurkat cells ＋ PTL 4 μmol / L group,BMSCs ＋ Jurkat cells ＋ PTL 8 μmol / L group,BMSCs ＋ Jurkat cells ＋ PTL12 μmol / L group,BMSCs ＋ Jurkat cells ＋ PTL 16 μmol / L group were setted. Adhesion rates were detected by adhesion test at hours 24,48 after the cultures,the expression of soluble vascular cell adhesion molecule- 1（ s VACM- 1） by ELISA,relative expression of BMSCs VCAM-1 mRNA by RT- PCR in all groups. Results At hours 24,48 after the cultures,adhesion rate was negatively correlated with PTL concentration in all groups（ r =- 0. 95,- 0. 91,P〈0. 05）. There was difference in s VCAM-1 expression in all groups（ P〈0. 05）,higher in groups BMSCs ＋ Jurkat cells,BMSCs ＋ Jurkat cells ＋ PTL 4 μmol / L,BMSCs ＋ Jurkat cells ＋ PTL 8 μmol / L,BMSCs ＋ Jurkat cells ＋ PTL 12 μmol / L than in BMSCs group（ P〈0. 05）,lower in PTL groups than in BMSCs ＋ Jurkat cells group（ P〈0. 05）. Expression level of s VCAM-1 was negatively correlated with PTL concentration in PTL groups（ r =- 0. 944,P = 0. 001）. There was difference in BMSCs VCAM-1 mRNA expression in each group（ P〈0. 05）,no difference between BMSCs group and BMSCs ＋ Jurkat cells ＋ PTL 16 μmol / L group. Expression level of BMSCs VCAM-1 mRNA was negatively correlated with PTL concentration in PTL groups（ r =- 0. 944, P = 0. 001）.Conclusion PTL inhibits the adhesion of BMSCs to Jurkat cells by decreasing VCAM- 1 expression. Weakened bone marrow microenvironment is of protective effect on leukemic cells.
出处
《中国全科医学》
CAS
CSCD
北大核心
2015年第5期535-539,共5页
Chinese General Practice
基金
广西自然科学基金资助项目(2012GXNSFAA053112
2014GXNSFAA118170)
