姜黄素联合顺铂通过内质网应激信号通路抑制腺样囊性癌ACC-M细胞增殖作用的研究

Inhibitory effect of curcumin combined with cisplatin on proliferation of ACC-M cells in adenoid cystic carcinoma through endoplasmic reticulum stress signaling pathway

目的探讨姜黄素与顺铂联合使用对人腺样囊性癌细胞(ACC-M)的抗肿瘤作用及其可能机制。方法使用不同浓度的姜黄素(5μM、10μM、20μM、40μM)单独或联合顺铂(8μM)处理腺样囊性癌ACC-M细胞24 h后,使用噻唑蓝和乳酸脱氢酶试剂盒检测细胞存活率和细胞毒性。采用20μM姜黄素、8μM顺铂及20μM姜黄素+8μM顺铂进行药物干预,并设立对照组(0.1%DMSO),24 h后倒置显微镜下观察各种细胞形态,蛋白质印迹法检测各组GRP78、p-elF2α、CHOP、p-JNK蛋白的表达水平。结果联合用药组的ACC-M细胞生长存活率均低于姜黄素组、顺铂组(P<0.05);联合组乳酸脱氢酶释放量均高于姜黄素、顺铂组(P<0.05)。与对照组相比,姜黄素组、顺铂组、联合用药组的GRP78、p-elF2α、CHOP表达水平升高(P<0.05)。联合用药组GRP78、p-elF2α、CHOP、p-JNK蛋白水平高于姜黄素组、顺铂组(P<0.05)。结论与单独用药相比,姜黄素联合顺铂能更有效地抑制ACC-M增殖,其机制可能与上调内质网应激相关蛋白GRP78、p-elF2α、CHOP、p-JNK的表达有关。

Objective To explore the anti-tumor effect of curcumin and cisplatin on human adenoid cystic carcinoma cell line(ACC-M)and the possible mechanism.Methods After treating ACC-M cells with different concentrations of curcumin(5μM,10μM,20μM,40μM)alone or in combination with cisplatin(8μM)for 24 hours,we detected the viability and cytotoxicity of these cells by MTT assay and lactate dehydrogenase kits.20μM curcumin,8μM cisplatin and 20μM curcumin+8μM cisplatin were used as medicine intervention,and a control group(0.1%DMSO)was established.Twenty-four hours later,we observed the cell morphology under inverted microscope and detected the protein expressions of GRP78,p-elF2α,CHOP and p-JNK by Western blotting.Results The survival rate of ACC-M cells in the combined-medicine group was lower than that in the curcumin group as well as that of the cisplatin group(P<0.05),while the release of lactate dehydrogenase of the combined-medicine group was higher than that of the curcumin group as well as that of the cisplatin group(P<0.05).Compared with those in the control group,the expressions of GRP78,p-elF2αand CHOP in the curcumin group,the cisplatin group and the combined-medicine group all increased(P<0.05).The protein levels of GRP78,p-elF2α,CHOP and p-JNK in the combined-medicine group were higher than those in the curcumin group and those in the cisplatin group(P<0.05).Conclusion Compared with treatment with single medicine,curcumin combined with cisplatin could inhibit ACC-M proliferation more effectively.The mechanism may be related to the up-regulation of GRP78,p-elF2α,CHOP,and p-JNK proteins related to endoplasmic reticulum stress.