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

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。

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

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 MTr 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 microg/ml to 640 microg/ 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 pjg/ml cerulenin 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.