羟基红花黄色素A逆转卵巢癌耐药细胞株A2780/DDP的作用及机制

摘要

目的 探讨羟基红花黄色素A(HSYA)对人卵巢癌A2780/DDP细胞株顺铂耐药性的逆转作用及作用机制.方法 培养A2780和A2780/DDP细胞,A2780组命名为正常组,A2780/DDP组分成对照组、顺铂组、HSYA组和联合组,顺铂组中加入25μg/ml DDP,HSYA组加入1mg/ml HSYA,联合组加入25μg/ml DDP和1mg/ml HSYA,正常组和对照组加入等体积培养基.采用MTT比色法、RTCA法和流式细胞检测法分别测定对照组、顺铂组和联合组的A2780/DDP细胞存活率、迁移能力和凋亡能力;采用Western blot检测正常组、对照组和HSYA组细胞的Erk1/2、p-Erk1/2、E-cadherinl、Vimentin、Snail和Twist表达含量.结果 联合组细胞存活率、细胞迁移能力较对照组和顺铂组下降(P<0.01),细胞凋亡率较对照组和顺铂组上升(P<0.01).Western blot结果示,与正常组比较,对照组Erk1/2、p-Erk1/2、Vimentin、Snail和Twist表达明显增加,E-cadherinl表达减少(P<0.05);与对照组比较,HSYA组Erk1/2、p-Erk1/2、Vimentin、Snail和Twist表达下降, E-cadherinl 表达增加(P<0.05).结论 HSYA能抑制Erk1/2、p-Erk1/2的表达,干预MAPK/ERK通路,进一步影响E-cadherinl、Vimentin、Snail和Twist表达,抑制EMT,从而逆转人A2780/DDP细胞对DDP的耐药,有效提高化疗敏感性.%Objective To investigate the reversal effect of hydroxysafflor yellow A(HSYA) on cisplatin resistance in human ovarian cancer cells A2780/DDP and its specific mechanism. Methods A2780 and A2780/DDP cells were cultured. The A2780/DDP group was named as normal group while the A2780/DDP group was divided into control group,DDP group,HSYA group and combined group. DDP group was added 25μg/ml DDP,HSYA group was added 1 mg/ml HSYA,and combined group was added 25μg/ml DDP and 1mg/ml HSYA,rest groups were conducted with equal volume medium. The survival rate,migration ability and apoptosis ability of A2780/DDP cells in control group,DDP group and combined group were measured by MTT colorimetry,RTCA and flow cytometry respectively. Western blot was used to detect the expression levels of Erk1/2, p-Erk1/2,E-cadherinl,Vimentin,Snail and Twist in normal group,control group and HSYA group respectively. Results Compared with control group and DDP group,A2780/DDP cells' survival rate and migration ability in combined group decreased significantly(P<0.01),the apoptosis ability increased (P<0.01). Western blot results showed that compared with normal group,the expressions of Erk1/2,p-Erk1/2,Vimentin, Snail and Twist in the control group were significantly increased while E-cadherinl was decreased (P<0.05). In relation to the control group, the expressions of Erk1/2,p-Erk1/2,Vimentin,Snail and Twist in HSYA group were decreased,and E-cadherinl was increased (P<0.05). Conclusion HSYA can inhibit the expression of Erk1/2 and p-Erk1/2, interfere with the MAPK/ERK pathway, further affect the expression of E-cadherinl, Vimentin, Snail and Twist, and inhibit EMT, thus reversing the resistance of human A2780/DDP cells to DDP and effectively improving the chemosensitivity.