脂肪酸合成酶抑制剂抑制人多发性骨髓瘤细胞增殖及诱导其凋亡的研究

Study on proliferation inhibiting and apoptosis inducing effects of cerulenin on multiple myeloma cells

目的明确脂肪酸合成酶(FAS)在人多发性骨髓瘤(MM)细胞中的高表达;探讨 FAS 抑制剂抑制 MM 细胞增殖及诱导其凋亡的机制。方法通过 RT-PCR 方法检测 FAS 在 MM 细胞系U266、RPMI8226细胞中的表达;以 MM 细胞系 U266细胞为模型,MTT 法观察 FAS 抑制剂浅蓝菌素(cerulenin)对 U266细胞增殖抑制率;以流式细胞术检测经 cerulenin 处理后 U266细胞 Annexin V 的表达及细胞周期变化。结果 MM 细胞系 U266、RPMI8226细胞均有 FAS mRNA 的高表达,而健康献血员外周血单个核细胞(PBMNC)中不表达 FAS mRNA;cerulenin 对 U266细胞增殖有显著的抑制作用,并呈剂量-效应相关;20μg/ml 的 cerulenin 作用于 U266细胞12h,细胞早期凋亡率为56.9%,24h 细胞早期凋亡率为69.3%,而对照组分别为4.3%和1.8%(P<0.01);细胞周期 DNA 分析发现,20μg/mlcerulenin 作用于 U266细胞12h,S 期细胞从对照组的9.7%上升至20.3%,作用24h,S 期细胞上升为29.8%。结论 FAS 在 MM 细胞中高表达;FAS 抑制剂可抑制 U266细胞的增殖;DNA 合成阻止在 S期,其抑制增殖的作用可能是通过促进 U266细胞早期凋亡;FAS 可能是一个潜在的抗 MM 的靶位。

Objective To determine whether fatty acid synthase （FAS） is expressed in human multiple myeloma（MM） cells and investigate the proliferation inhibition effect of fatty acid synthase inhibitor cerulenin on multiple myeloma cell line U266 and its mechanism. Methods FAS mRNA expression in human MM cell line U266, RPMI8226 cell was assayed by RT-PCR. The proliferation inhibition rate of U266 cells was assayed by MTT analysis. Cell apoptosis and cycle distribution were evaluated by flow cytometry （FCM）. Results FAS mRNA was highly expressed in human multiple myeloma cell lines as compared with healthy donor PBMNCs. After U266 cells were treated with cerulenin （ the concentrations from 5 μg/ml to 640 μg/ ml） for 24 h, the cell proliferation was markedly inhibited with a dose related manner, while the inhibition rate of human skin fibroblast cells were all lower than 30%. When U266 cells were treated with 20 μg/ml eerulenin for 12 h and 24 h,the early apoptosis rate revealed by Annexin V/PI were 56.9% and 69.3% respectively, being higher than that of the blank controls （4.3% and 1.8%, P 〈0.01 ）. Cell cycle analysis showed it was blocked in S phase. Conclusion FAS is highly expressed in human MM. Cerulenin could induce apoptosis and inhibit proliferation of U266 cells. FAS might be a new potential target for multiple myeloma treatment.