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目的探讨和厚朴酚(HNK)及硼替佐米对人多发性骨髓瘤KM3细胞增殖的影响及诱导凋亡作用。方法本实验分实验组和对照组(A组),实验组分为HNK单药组(B、C、D、E、F、G组的药物质量浓度分别为4、6、8、10、12、15μg/mL)、硼替佐米单药组(H、I、J组的药物质量浓度分别为5、10、20ng/mL)及二者联合用药组(K组为HNK4μg/mL+硼替佐米5ng/mL,L组为HNK4μg/mL+硼替佐米10ng/mL,M组为HNK4μg/mL+硼替佐米20ng/mL)。在不同的时间(24、48、72h)采用MTT比色法检测对照组和实验组细胞增殖活性;采用流式细胞术检测对照组和实验组细胞凋亡的变化。结果和厚朴酚4～15μg/mL对KM3细胞作用24、48、72h的细胞增殖抑制率分别由20.38%升至80.54%、27.45%升至89.99%、44.94%升至90.91%,不同组、不同作用时间相比差异均有统计学意义(P<0.05)。H组、I组、J组KM3细胞48h的细胞增殖抑制率分别是13.13%、36.22%、53.99%,各组间差异有统计学意义(P<0.05)。K组、L组、M组作用48h的细胞增殖抑制率分别是52.68%、69.99%、78.53%,K组与H组、L组与I组、M组与J组比较差异有统计学意义(P<0.05)。流式结果显示B组、C组、E组作用于KM3细胞24h和48h后细胞凋亡率分别是6.92%、16.15%、60.70%和18.84%、37.21%、86.07%,同一时间不同实验组之间差异有统计学意义(P<0.05)。H组、I组作用于KM3细胞24、48h的细胞凋亡率分别是9.27%、17.87%和11.92%、53.51%,同一时间不同实验组之间差异有统计学意义(P<0.05)。K组、L组作用于KM3细胞24、48h,细胞凋亡率分别是15.75%、22.18%和35.96%、74.70%,K组、L组细胞凋亡率明显高于H组、I组,差异有统计学意义(P<0.05)。结论和厚朴酚与硼替佐米均可诱导KM3细胞凋亡,抑制KM3细胞的增殖,两者联合作用能显著提高KM3细胞的凋亡率,增强抑制KM3细胞的生长。
Objective To investigate the effects of honokiol (HNK) and bortezomib on the proliferation of human multiple myeloma KM3 cells and the induction of apoptosis. Methods The experimental group was divided into experimental group and control group (group A). The experimental group consisted of HNK single drug group (group B, C, D, E, F, G, drug quality concentrations were 4,6,8,10,12 , 15μg / mL), bortezomib monotherapy group (drug concentrations of 5, 10 and 20ng / mL in H, I and J groups, respectively) and combination group (HNK4μg / mL and bortezomib in group K / mL, L group was HNK4μg / mL + bortezomib 10ng / mL, M group was HNK4μg / mL + bortezomib 20ng / mL). At different time (24,48,72h), MTT assay was used to detect the cell proliferation activity in control group and experimental group. The changes of apoptosis in control group and experimental group were detected by flow cytometry. Results The inhibitory rates of 4 ~ 15μg / mL honokiol on KM3 cells were increased from 20.38% to 80.54%, from 27.45% to 89.99% and from 44.94% to 90.91%, respectively. There were significant differences in different time of action (P <0.05). The inhibitory rates of KM3 cells in H group, I group and J group at 48h were 13.13%, 36.22% and 53.99%, respectively. The differences among the three groups were statistically significant (P <0.05). The inhibitory rates of cell proliferation of K group, L group and M group at 48h were 52.68%, 69.99% and 78.53% respectively. There was significant difference between K group and H group, L group and I group, M group and J group P <0.05). The results of flow cytometry showed that the apoptotic rates of KM3 cells were 6.92%, 16.15%, 60.70% and 18.84%, 37.21% and 86.07% respectively after treated with B, C and E groups at different time points The difference was statistically significant (P <0.05). The apoptotic rates of H group and I group on KM3 cells at 24 h and 48 h were 9.27%, 17.87% and 11.92%, 53.51%, respectively. There was significant difference between the experimental groups at the same time (P <0.05). The apoptotic rates of KM3 cells in K group and L group were 15.75%, 22.18% and 35.96%, 74.70% respectively at 24 and 48 hours. The apoptosis rates in K and L groups were significantly higher than those in H and I groups There was statistical significance (P <0.05). Conclusion Both honokiol and bortezomib can induce the apoptosis of KM3 cells and inhibit the proliferation of KM3 cells. The combination of both can significantly increase the apoptosis rate of KM3 cells and increase the inhibition of the growth of KM3 cells.