二十碳五烯酸抑制自噬反应提高膀胱癌细胞化疗敏感性  

Eicosapentaenoic acid enhances the sensitivity of bladder cancer cell line T24 to cisplatin via inhibiting autophagy reaction

作　　者：张建育[1] 高瑞林[1] 李毅宁[1] 孙建国[1] 郭一泓[1] Zhang Jianyu;Gao Ruilin;Li Yining;Sun Jianguo;Guo Yihong(Department of Urology,the Second Affiliated Hospital,Medical University of Fujian,Quanzhou 362001, China)

机构地区：[1]福建医科大学附属第二医院泌尿外科,泉州362001

出　　处：《中华实验外科杂志》2018年第12期2202-2204,共3页Chinese Journal of Experimental Surgery

基　　金：福建省自然科学基金(2018J01280);泉州市科技计划项目基金(2015257)。

摘　　要：目的探讨二十碳五烯酸(EPA)对膀胱癌T24细胞顺铂(DDP)化疗敏感性的影响及其机制。方法根据不同药物干预情况分为5组:对照组、EPA组,DDP组、EPA+DDP组、EPA+DDP+雷帕霉素组(EPA+Rapa+DDP组)。细胞计数(CCK-8)试剂盒法和流式细胞仪检测T24细胞增殖抑制率和凋亡率,Westernblot法检测凋亡因子[活化的天冬氨酸特异性半胱氨酸蛋白酶3(CleavedCaspase-3)、B细胞淋巴瘤/白血病-2相关X蛋白(bax)],抗凋亡因子(bcl-2)和自噬反应相关因子(LC3-Ⅱ、Beclin-1)的表达。结果与DDP组比较,EPA+DDP组T24细胞的增殖抑制率明显增高[(73.19±6.92)%比(48.62±4.0)%,P=0.000],凋亡率明显增高[(64.30±6.53)%比(35.90±4.21%,P=0.010]。Westernblot检测显示,与DDP组比较,EPA+DDP组CleavedCaspase-3、bax表达增加,而bcl-2、LC3-Ⅱ、Beclin-1蛋白表达减少(P均=0.000)。EPA+Rapa+DDP组增殖抑制率和凋亡率较EPA+DDP组明显降低,同时自噬反应表达增加。结论EPA预处理增强DDP抑制膀胱癌T24细胞增殖,促进细胞凋亡,提高DDP化疗敏感性,其机制可能是通过抑制化疗后增高的自噬反应。Objective To investigate the effect and underlying mechanism of eicosapentaenoic acid (EPA) on the sensitivity of bladder cancer cell line T24 to cisplatin. Methods T24 cells were randomly divided into five groups: control group, EPA group, cisplatin group (group DDP), EPA+ DDP group and rapamycin group (group EPA+ Rapa+ DDP). Cell counting Kit-8 (CCK-8) method was used to evaluate inhibition ratio of cell proliferation. The apoptotic ratio was examined by flow eytometry (FCM). The protein expressions of apoptosis cytokines [cysteine-containing aspartate-specific proteases (Cleaved Caspase)-3, B cell lymphoma/leukemia-2 associated X protein (bax), B cell lymphoma/leukemia-2 (bcl-2)] and autophagy reaction cytokines (LC3-Ⅱ, Beclin-1) were detected by Western blotting. Results Compared to DDP treatment alone, the proliferation inhibition ratio and apoptotic ratio of T24 cells were enhanced by EPA+ DDP treatment [(73.19±6.92)% vs. (48.62±4.80)%, P=0.000; (64.30±6.53)% vs. (35.90±4.21)%, P=0.010, respectively]; Western blotting showed that the expressions of apoptotic factors were increased by EPA+ DDP treatment. Meanwhile, the protein expressions of bcl-2, LC3-Ⅱ and Beclin-1 were significantly inhibited by EPA+ DDP treatment (P=0.000). However, the salutary effects of EPA would been reversed by autophagy reaction activation rapamycin. Conclusion EPA enhances the sensitivity of bladder cancer cell line T24 to cisplatin, which may suppress activation of autophagy reaction.

关 键 词：膀胱癌细胞 二十碳五烯酸 顺铂 自噬反应 

分 类 号：R737.14[医药卫生—肿瘤]