冬凌草甲素对多发性骨髓瘤抗肿瘤的机制研究

本研究探讨冬凌草甲素（Oridonin）对人多发性骨髓瘤（MM）细胞株U266抗肿瘤作用及其分子机制。CCK-8法检测冬凌草甲素对细胞增殖的影响；瑞氏染色法观察细胞的形态学改变；流式细胞术检测细胞凋亡；实时荧光定量PCR方法检测U266细胞FGFR3、BC12、CCNDJ、MYC基因表达水平的变化；Westernblot方法分析BCL2、MYC、CCNDl、FGFR3和P53蛋白表达水平的变化。结果表明，①冬凌草甲素能够抑制U266细胞增殖，且呈浓度和时间依赖性；②10μmol／L冬凌草甲素处理U266细胞24h后，光学显微镜下可观察到典型的细胞凋亡形态学改变；③U266细胞凋亡比例随冬凌草甲素药物浓度及作用时间的延长而增加；④经冬凌草甲素处理后的U266细胞中，FGFR3、BCL2、CCNDJ、MYC基因表达下调，同时其相应的蛋白质表达下调，P53蛋白表达上调，上述变化均呈浓度和时间依赖性。结论：冬凌草甲素可以抑制人多发性骨髓瘤U266细胞增殖并诱导其凋亡，发挥其抗肿瘤作用。本研究为MM新的靶向治疗方案选择提供理论依据。

Mechanism Concerning Antitumor Effect of Oridonin on Multiple Myeloma Cell Line U266

This study was purposed to investigate the antitumor effect of oridonin on human multiple myeloma cell line U266 and its possible mechanism. The CCK-8 test was used to determine the inhibitory effect of oridonin on prolif- eration of U266 cells. The morphological changes of U266 cells were observed under optical microscope. The apoptosis rate of U266 cells was detected by flow cytometry. The mRNA levels of FGFR3, BCL2, CCND1 and MYC genes were quantified by using real-time quantitative PCR method, and the protein levels of BCL2, MYC, CCND1, FGFR3 and P53 were detected by Western blot. The results showed that the oridonin obviously inhibited the growth of U266 cell in dose-and time-dependent manners. As for morphological changes, characteristic apoptotic cells presented in U266 cells treated with 10 μmol/L oridonin for 24 hours. The apoptotic rate of U266 ceils increased in dose and time dependent manners; after treatment of U266 cells with oridonin the mRNA levels of FGFR3, BCL2, CCND1 and MYC as well as the their protein levels decreased. Occasionally, the oridonin up-regulated the protein levels of P53 in the same manner. It is concluded that the oridonin can exert its anti-tumor effect by inhibiting proliferation and inducing apoptosis of U266 cell in dose dependent and time dependent manners, that maybe give the clues about new program of target therapy for multiple myeloma.