摘要
目的:探讨丙戊酸钠(valproic acid sodium,VPA)对人骨髓瘤RPMI8226和U266细胞增殖及IL-6/JAK/STAT信号通路的作用。方法:RPMI8226和U266细胞经不同浓度VPA处理12及24 h后进行实验,MTT法检测VPA对细胞增殖的影响,流式细胞术检测VPA处理后的细胞周期及凋亡率,ELISA法检测培养细胞上清IL-6的浓度,半定量RT-PCR检测VPA作用后RPMI8226和U266细胞中STAT3、STAT5、Bcl-xL、Mcl-1、c-Myc、CCND1和VEGF mRNA表达水平。Western blotting检测JAK2和STAT5总蛋白及磷酸化蛋白表达水平。结果:(1)10、20及40μmol/L VPA处理RPMI8226和U266细胞12 h及24 h后,细胞存活率值显著下降,与各对照组比较均有显著差异(P<0.05),各对照组之间无显著差异。(2)VPA处理RPMI8226和U266细胞24 h后,G1/G0期细胞增多,S期细胞减少,细胞凋亡率增加,且凋亡率与VPA的作用剂量相关。(3)VPA处理RPMI8226和U266细胞24 h后,上清IL-6浓度与VPA浓度呈负相关。(4)RPMI8226和U266细胞组在40μmol/L VPA处理后其STAT3、STAT5、Bcl-xL、Mcl-1、c-Myc、CCND1和VEGF mRNA表达减低,与未处理组比较有显著差异。(5)与对照组比较,VPA处理组p-JAK2、JAK2、p-STAT5和STAT5蛋白表达均显著降低。结论:(1)VPA对多发性骨髓瘤细胞有体外抑制作用;(2)VPA对骨髓瘤细胞株的体外抑制可能是通过调节IL-6/JAK/STAT信号通路来实现的。
AIM ： To investigate the effects of sodium valproate （VPA） on the proliferation of multiple myelo- ma cell lines RPMI8226 and U266 and the regulation of IL-6/JAK/STAT signaling pathway. METHODS： The cells were treated with different concentrations of VPA for 12 h and 24 h. The growth of RPMI8226 cells and U266 cells was detected by MTT assay. Apoptotic rates and cell cycle were analyzed by flow cytometry. The mRNA expression of STAT3, STAT5 and STAT target genes Bcl-xL, Mcl-1, c-Myc, CCND1 and VEGF was measured by RT-PCR. Western blotting analysis was used to determine the total proteins and protein phosphorylation levels of JAK2 and STAT5. RESULTS： VPA inhibited the growth and induced the apoptosis of RPMI8226 cells and U266 cells in a concentration- and time-dependent manner. The levels of IL-6 in the culture supernatants of RPMI8226 cells and U266 cells treated with VPA were significantly higher than that in negative control group. VPA down-regulated the mRNA expression of STAT3, STATS, Bcl-xL, Mcl-1, c-Myc, CCND1 and VEGF. After treated with VPA, the protein levels of p-JAK2, JAK2, p-STATS and STATS in RPMI8226 cells and U266 cells were significantly lower than those in control group. CONCLUSION： VPA inhibits the proliferation of PR- MI8226 cells and U266 cells in vitro. The modulation of IL-6/JAK/STAT signaling pathway may be involved in its potential mechanisms.
出处
《中国病理生理杂志》
CAS
CSCD
北大核心
2013年第5期833-838,共6页
Chinese Journal of Pathophysiology
基金
温州市科技计划(No.Y20100106)
