氧化苦参碱通过P38MAPK通路抑制人宫颈癌HeLa细胞增殖

Inhibitory effect of oxymatrine on human cervical carcinoma He La cell proliferation through P38 MAPK signaling pathways

目的观察氧化苦参碱(OXY)对人宫颈癌HeLa细胞增殖的影响,并探讨其机制及作用。方法人宫颈癌HeLa细胞经过系列浓度的OXY(1.0、2.0和4.0mg/ml)处理不同时间后,采用噻唑蓝(MTT)法测定细胞增殖的变化。免疫印迹(Westernblot)方法测定OXY对HeLa细胞PCNA、CyclinD1和P38MAPK、P-P38MAPK计学意义表达的影响。并采用AnnexinV-PI双染测定细胞凋亡。结果氧化苦参碱成浓度时间依赖性抑制HeLa细胞增殖;高浓度氧化苦参碱(4.0mg/ml)不仅抑制HeLa细胞增殖,而且促进细胞凋亡。与对照组比,OXY处理细胞的PCNA和CyclinD1蛋白表达明显降低(P<0.05,P<0.01),而P-P38MAPK蛋白表达水平明显升高;P38MAPK特异性抑制剂可缓解OXY对HeLa细胞增殖的抑制率,增加PCNA和CyclinD1蛋白表达。结论OXY能够通过提高P38MAPK磷酸化而下调PCNA和CyclinD1表达,抑制HeLa细胞增殖。

[ Objective ] To investigate the inhibitory effect of oxymatrine （OXY） on the human cervical carcinoma HeLa cell proliferation through the P38 MAPK signaling pathways. [Methods] HeLa cells were cultured in RPMI 1640 medium. MTT assay was used to determine cell proliferation. PCNA, Cyclin D1, P38 MAPK, P-P38 MAPK protein expressions were tested by western blot. Apoptosis was detected by Annexin V-PI double staining. [ Results ] OXY inhibited the proliferation of HeLa cells in a concentration and time dependent manner. Compared with control group, the expressions of PCNA, Cyclin D1 protein were significantly lower in OXY group （P 〈 0.05, P 〈 0.01）, and the expression of P-P38 MAPK was significantly higher in OXY group （P 〈 0.01）. The expression of PCNA, Cyclin D1 were significantly higher, while P-P38 MAPK were significantly lower in OXY ＋ SB203580 group than those in OXY group （P 〈 0.01）. [Conclusions] OXY can inhibit HeLa cell proliferation and down-regulate PCNA and Cyclin D1 expressions by activating P-P38 MAPK pathway.