苦瓜素Ⅰ抑制乳腺癌MCF-7/DOX细胞多柔比星耐药的研究  

Suppression of momordicin Ⅰ on the resistance of breast cancer MCF-7/DOX cells to doxorubicin

作　　者：汤喜兰 李洪铭 梁新丽 谢冰斌[3] 熊印华 邱雨美 谢梦蝶 刘志颖 董伟 TANG Xi-lan;LI Hong-ming;LIANG Xin-li;XIE Bing-bin;XIONG Yin-hua;QIU Yu-mei;XIE Meng-die;LIU Zhi-yin;DONG Wei(School of Pharmacy,Jiangxi Science&Technology Normal University,Jiangxi Nanchang 330013,China;Key Laboratory of Modern Preparation of Chinese Medicine,Ministry of Education,Jiangxi University of Traditional Chinese Medicine,Jiangxi Nanchang 330004,China;Department of Otorhinolaryngology,Head&Neck Surgery,Second Affiliated Hospital,Nanchang University,Jiangxi Nanchang 330006,China)

机构地区：[1]江西科技师范大学药学院,江西南昌330013 [2]江西中医药大学现代中药制剂教育部重点实验室,江西南昌330004 [3]南昌大学第二附属医院耳鼻咽喉头颈外科,江西南昌330006

出　　处：《中国医院药学杂志》2021年第13期1293-1298,共6页Chinese Journal of Hospital Pharmacy

基　　金：国家自然科学基金项目(编号:82060733,81660173,81660692);江西中医药大学现代中药制剂教育部重点实验室开放项目(编号:TCM-201911);江西科技师范大学博士启动基金项目(编号:2017BSQD017)。

摘　　要：目的:研究苦瓜素Ⅰ(momordicin Ⅰ,MMI)对人乳腺癌耐多柔比星(doxorubicin,DOX)MCF-7/DOX细胞耐药的影响及其作用机制。方法:MTT比色法检测MMI对MCF-7/DOX细胞的毒性,考察MMI对MCF-7/DOX细胞多柔比星敏感性的影响;实时定量PCR和western blot方法检测DOX单独以及MMI与DOX联用后,MCF-7/DOX细胞中MDR1在mRNA和蛋白质水平的表达变化;进一步采用实时定量PCR方法检测DOX单独以及MMI与DOX联用后,MCF-7/DOX细胞中与甘油磷脂代谢相关的AGPAT2、CEPT1、CHKA、DGKA、PCYT1A、PLA2G15等基因表达的变化。结果:MMI(12.5,25,50μg·mL^(-1),)可明显抑制MCF-7/DOX细胞的增殖,MMI对MCF-7/DOX细胞的最大无毒浓度为6.25μg·mL^(-1)。MMI 6.25μg·mL^(-1)与DOX联用后,DOX对MCF-7/DOX细胞的半数抑制浓度(IC50)下降,MMI使得MCF-7/DOX细胞对DOX的敏感性提高了13.88倍。与DOX 50μg·mL^(-1)单独组相比,MMI 6.25μg·mL^(-1)与DOX 50μg·mL^(-1)联用后,MCF-7/DOX细胞中的MDR1 mRNA显著下调(P<0.05),MDR1蛋白表达有下降趋势。进一步研究发现MMI 6.25μg·mL^(-1)与DOX 50μg·mL^(-1)联用可协同下调MCF-7/DOX细胞甘油磷脂代谢相关基因AGPAT2、CEPT1、CHKA、DGKA、PCYT1A、PLA2G15等表达。结论:MMI可增强乳腺癌MCF-7/DOX细胞多柔比星敏感性,抑制MDR1表达,逆转耐药,此作用可能与其抑制甘油磷脂代谢有关。OBJECTIVE To explore the effect of momordicin I(MMI)on the drug resistance of human breast cancer MCF-7/DOX cells and elucidate its mechanism.METHODS MTT method was utilized for detecting the toxicity of MMI on MCF-7/DOX cells and examining the effect of MMI on cellular sensitivity to doxorubicin(DOX)chemotherapy.Real-time quantitative polymerase chain reaction(PCR)and Western blot were employed for observing the changes of MDR1 mRNA level and protein expression after treatments with DOX in the presence or absence of MMI.Real-time quantitative PCR method was employed for measuring the mRNA levels of AGPAT2,CEPT1,CHKA,DGKA,PCYT1A and PLA2G15 after a treatment of DOX in the presence or absence of MMI.RESULTS Cytotoxicity assays showed that MMI(12.5/25/50μg·mL^(-1))obviously inhibited cellular growth and the maximal non-toxic concentration of MMI was 6.25μg·mL^(-1).As compared with DOX alone group,IC50 value of DOX decreased after a treatment of MMI 6.25μg·mL^(-1) plus DOX.MMI enhanced cellular sensitivity to DOX by 13.88 folds.As compared with DOX 50μg·mL^(-1) alone group,MDR1 mRNA level was significantly down-regulated(P<0.05)and MDR1 protein expression trended downward after a treatment of MMI 6.25μg·mL^(-1) plus DOX 50μg·mL^(-1).Furthermore,a combination of MMI 6.25μg·mL^(-1) and DOX 50μg·mL^(-1) synergistically down-regulated the mRNA levels of AGPAT2,CEPT1,CHKA,DGKA,PCYT1A and PLA2G15.CONCLUSION MMI can enhance the sensitivity of breast cancer MCF-7/DOX cells to DOX,suppress MDR1 expression and reverse drug resistance.It may be correlated with its inhibition of glycerophospholipid metabolism.

关 键 词：苦瓜素Ⅰ 乳腺癌 多柔比星 耐药 脂质代谢 

分 类 号：R966[医药卫生—药理学]