摘要
目的探讨小白菊内酯（parthenolide，PTL）对多发性骨髓瘤（multiple myeloma，MM）细胞蛋白酶体活性及白细胞介素-6（interleukin-6，IL-6）表达的影响，以期了解PTL抗MM的分子机制。方法2006年5月至2007年3月华中科技大学同济医学院附属协和医院血液病研究所，体外培养人MM细胞系PRMI8266，与不同浓度的PTL作用不同时间。以荧光底物法检测细胞蛋白酶体活性，逆转录聚合酶链反应（RT-PCR）检测IL-6基因表达，酶联免疫吸附试验（ELISA）法检测MM细胞培养上清中IL-6的质量浓度。结果2~10μmol/L的PTL作用16h对PRMI8266细胞蛋白酶体的糜蛋白酶活性具有明显抑制作用，其效应呈现浓度依赖性；10μmol/L可达到接近50%的活性抑制。RT-PCR检测结果表明，2，5，10μmol/L的PTL作用24h，MM细胞的IL-6基因 mRNA表达强度与对照相比均明显降低；2，5，10μmol/L的PTL作用MM细胞48h后，培养上清中IL-6质量浓度分别为（92.6±4.3）ng/L、（67.1±5.7）ng/L、（43.5±4.9）ng/L，与对照组（148.7±8.2）ng/L相比，差异有显著性意义（P〈0.01）。结论PTL能明显抑制PRMI8266细胞的蛋白酶体活性，降低IL-6基因表达，减少MM细胞IL-6的自分泌。提示PTL抑制蛋白酶体活性及IL-6表达可能是其抗MM的机制之一。
Objective To investigate the effect of parthenolide（PTL） on activity of proteasome and expression of IL-6 of human multiple myeloma（MM） cells so as to understand the molecular mechanisms of anti-MM effect of PTL. Methods After treatment with different concentrations of PTL at different time points, the proteasomal activity of RPMI8226 cells was measured by detecting the fluorescence units given out by substrate,and the expression of interleukin-6 was evaluated by RT-PCR and the concentrations of interleukin-6 of the supernant were measured by ELISA. Results The cells were cultured with PTL at 2 to 10 μmol/L. Sixteen hours later,significant inhibited proteasomal activities could be observed in a dose-dependent manner. When treated with PTL at 10μmol/L,the proteasomal activity could be inhibited to 50 percent. PTL at 2,5 and 10 μmol/L significantly inhibited the expression of IL-6 at mRNA level. After 48-hour-treatment with PTL at different doses of 2 to 10 μmol/L, concentrations of IL-6 in the supernant were 92. 6 ± 4. 3,67.1 ± 5.7 and 43.5±4. 9 ng/L（ compared with 148.7 ± 8.2 for the control,P 〈 0. 01 ）. Conclusion PTL can significantly inhibit the proteasomal activity and expression and secretion of IL-6 of MM cells,which seems to be one of the mechanisms of anti-MM effects of PTL. It is the first report of PTL inhibiting the proteasomal activity of MM cells.
出处
《中国实用内科杂志》
CAS
CSCD
北大核心
2007年第18期1448-1450,共3页
Chinese Journal of Practical Internal Medicine
