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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 the proteasome activity and the expression of interleukin-6 (IL-6) in multiple myeloma (MM) Molecular mechanism. Methods From May 2006 to March 2007, Institute of Hematology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology. Human MM cell line PRMI8266 was cultured in vitro. The effect of PTMI was different from that of PTMI. The activity of cell proteasome was detected by fluorescent substrate method. The expression of IL-6 gene was detected by reverse transcription-polymerase chain reaction (RT-PCR). The quality of IL-6 in culture supernatant of MM cells was detected by enzyme linked immunosorbent assay (ELISA) concentration. Results PTL at the concentration of 2 ~ 10μmol / L significantly inhibited the chymotrypsin activity of proteasome in PRMI8266 cells in a concentration - dependent manner. At 10μmol / L, the inhibitory activity of PTL reached 50%. The results of RT-PCR showed that the expression of IL-6 mRNA in MM cells was significantly decreased at 2, 5 and 10μmol / L PTL for 24h, and the MM cells treated with 2,5,10μmol / L PTL at 48h The concentrations of IL-6 in the culture supernatant were (92.6 ± 4.3) ng / L, (67.1 ± 5.7) ng / L and (43.5 ± 4.9) ng / L, L, the difference was significant (P <0.01). Conclusion PTL can significantly inhibit the proteasome activity of PRMI8266 cells, decrease the expression of IL-6 gene and decrease the autocrine of IL-6 in MM cells. Tip PTL inhibition of proteasome activity and IL-6 expression may be one of its anti-MM mechanism.