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- 2015
蜂斗菜素诱导骨髓瘤RPMI 8226细胞凋亡及其机制
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Abstract:
摘要：目的 观察蜂斗菜素对骨髓瘤RPMI 8226细胞凋亡的影响，并探讨其作用机制。方法 采用锥虫蓝拒染法检测蜂斗菜素对RPMI 8226细胞的增殖抑制作用，TUNEL法检测蜂斗菜素诱导的RPMIS226细胞凋亡，Hochest 33258荧光染色法观察细胞核的变化，Western blot技术检测Caspase-3、8、9蛋白表达和MEK/ERK、p38MAPK蛋白磷酸化水平。结果 蜂斗菜素作用24、48、72h后对骨髓瘤RPMI 8226细胞均有显著的增殖抑制作用(P均<0.01)；蜂斗菜素呈时间和浓度依赖地诱导骨髓瘤RPMI 8226细胞凋亡(P<0.05，P<0.05)，Caspase抑制剂预处理可明显抑制细胞凋亡；蜂斗菜素作用72h后，Caspase-3、8、9蛋白表达显著增加(P<0.05，P<0.01，P<0.05)，ERK1/2及MEK蛋白磷酸化水平显著下降(P<0.01，P<0.05)，p38MAPK蛋白磷酸化水平显著上升(P<0.01)。结论 蜂斗菜素能够抑制骨髓瘤RPMI 8226细胞增殖，诱导细胞凋亡，其机制可能与Caspase-3、8、9蛋白激活，ERK/MEK及p38MAPK蛋白磷酸化改变有关。
ABSTRACT: Objective To investigate the apoptotic effect of petasin on myeloma RPMI 8226 cells and the mechanisms. Methods The inhibition of petasin on the proliferation of myeloma RPMI 8226 cells was tested by trypan blue assay. Apoptosis of RPMI 8226 cells was measured by terminal-deoxynueleotidyl transferase mediated dUTP nick end labeling (TUNEL) assay and Hoechst 33258 staining assay. Effects of petasin on caspase-3, 8 and 9 expressions, phosphorylation of ERK1/2, MEK(p-ERK1/2; p-MEK) and p38MAPK(p-p38MAPK) protein were analyzed by Western blot. Results Incubation by petasin for 24h, 48h or 72h could significantly inhibit the proliferation of myeloma RPMI 8226 cells (P<0.01, P<0.01, P<0.01). Petasin induced the apoptosis of myeloma RPMI 8226 cells in time- and concentration-dependent manners (P<0.05, P<0.05). Caspase inhibitor pretreatment could significantly inhibit the apoptosis of myeloma cells. After cultured with petasin for 72h, the expressions of caspase-3, 8 and 9 were obviously enhanced (P<0.05, P<0.01, P<0.05) and phosphorylation of p-p38MAPK of RPMI8226 cells was significantly increased (P<0.01). However, phosphorylation of p-ERK1/2 and p-MEK was decreased significantly (P<0.01, P<0.05). Conclusion Petasin can inhibit the proliferation of myeloma RPMI 8226 cells and induce apoptosis. The mechanism may be related to the activation of caspase-3, 8 and 9 proteins and the changes in phosphorylation of p38MAPK, ERK1/2 and MEK
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