Objective: To investigate the effect of celastrol on the proliferation and apoptosis of human multiple myeloma (MM) cell lines, reveal the relationship between IRAK4/ERK/p38 signaling pathway and celastrol regulating the proliferation and apoptosis of H929 and ARP-1 cells, and explore whether celastrol combined with bortezomib has synergistic effect.
Methods: CCK-8 method was used to detect the viability of MM cell lines H929 and ARP-1 treated by different concentrations of celastrol, bortezomib, and their combination, and the synergistic effect was determined by Kim's formula. The apoptosis rate of H929 cells and necrosis rate of ARP-1 were detected by Annexin V/PI method. The expression of key proteins and apoptosis proteins in IRAK4/ERK/p38 signaling pathway were detected by Western blot.
Results: Celastrol could significantly inhibit the proliferation of H929 and ARP-1 cells (r=0.9018, r=0.9244) and induce apoptosis in a time-dependent manner. Compared with the control group, celastrol could significantly up-regulate the expression of PARP and cleaved caspase-3 while down-regulate the expression of p-IRAK4, p-ERK, and p-p38 in H929 and ARP-1 cells. Celastrol and bortezomib alone inhibited the proliferation of H929 and ARP-1 cells. Compared with celastrol and bortezomib alone, their combination had lower cell survival rate and higher apoptosis rate (P<0.05).
Conclusion: Celastrol can inhibit the proliferation and promote the apoptosis of H929 and ARP-1 cells, which may be related to inhibiting the phosphorylation of IRAK4 and blocking the activation of IRAK4/ERK/p38 signaling pathway. Celastrol combined with bortezomib has synergistic effect, which can more effectively inhibit the proliferation and induce apoptosis of H929 and ARP-1 cells.
题目: 基于IRAK4/ERK/p38信号通路雷公藤红素对多发性骨髓瘤细胞增殖和凋亡的影响.
目的: 探讨雷公藤红素对人多发性骨髓瘤细胞增殖和凋亡的影响，并揭示IRAK4/ERK/p38信号通路与雷公藤红素调控H929、ARP-1细胞增殖和凋亡的关系，探究雷公藤红素联合硼替佐米是否具有协同作用.
方法: 采用CCK-8法检测多发性骨髓瘤细胞株H929、ARP-1细胞经不同浓度雷公藤红素、硼替佐米以及二者联用后的细胞活力，并利用金氏公式判定协同药效。Annexin V/PI法检测H929细胞凋亡率和ARP-1细胞坏死率。蛋白免疫印迹法检测雷公藤红素对IRAK4/ERK/p38信号通路中关键蛋白和凋亡蛋白表达的影响.
结果: 雷公藤红素能够呈时间-浓度依赖性地显著抑制H929、ARP-1细胞的增殖（r=0.9018，0.9244），并诱导细胞的凋亡；与对照组比较，雷公藤红素能够明显上调H929、ARP-1细胞内PARP、cleaved caspase-3表达，下调p-IRAK4、p-ERK、p-p38表达。雷公藤红素、硼替佐米单用对H929、ARP-1细胞均有增殖抑制作用，而联合用药与单药相比，前者细胞存活率更低，凋亡率更高（P<0.05）.
结论: 雷公藤红素能抑制H929和ARP-1细胞增殖、促进其凋亡，其机制可能与抑制IRAK4的磷酸化、阻断IRAK4/ERK/p38信号通路的激活有关；雷公藤红素联合硼替佐米具有协同作用，能更有效地抑制H929、ARP-1细胞增殖并诱导其凋亡.
Keywords: ARP-1; H929; IRAK4/ERK/p38 signaling pathway; bortezomib; celastrol; multiple myeloma.