辣椒素对多柔比星耐药的乳腺癌细胞的耐药逆转作用

doi:10.5246/jcps.2021.08.054

中国药学(英文版) ›› 2021, Vol. 30 ›› Issue (8): 666-674.DOI: 10.5246/jcps.2021.08.054

辣椒素对多柔比星耐药的乳腺癌细胞的耐药逆转作用

周毅¹^,²^,^*(), 王婷²^,³, 赵佳佳², 蒋学华²^,^*()

1. 重庆医科大学附属第一医院药学部, 重庆 400016
2. 四川大学华西药学院靶向药物及释药系统教育部重点实验室, 四川成都 610041
3. 四川省肿瘤医院药学部, 四川成都 610041

收稿日期:2021-01-12 修回日期:2021-02-20 接受日期:2021-03-11 出版日期:2021-08-29 发布日期:2021-08-29
通讯作者: 周毅, 蒋学华
作者简介:
+ Tel.: +86-23-89012410, E-mail: johnny@hospital.cqmu.edu.cn
+ E-mail: jxh1013@163.com
基金资助:
Chongqing Nature Science (Grant No. cstc2020jcyj-msxmX0858).

The reversal effect of capsaicin on doxorubicin-resistant breast cancer cells

Yi Zhou¹^,²^,^*(), Ting Wang²^,³, Jiajia Zhao², Xuehua Jiang²^,^*()

1 Department of Pharmacy, the First Affiliated Hospital of Chongqing Medical University, Chongqing 400016, China
2 Key Laboratory of Drug Targeting and Drug Delivery System, West China School of Pharmacy, Sichuan University, Chengdu, Sichuan 610041, China
3 Department of Pharmacy, Sichuan Cancer Hospital & Institution, Chengdu, Sichuan 610041, China

Received:2021-01-12 Revised:2021-02-20 Accepted:2021-03-11 Online:2021-08-29 Published:2021-08-29
Contact: Yi Zhou, Xuehua Jiang

摘要/Abstract

摘要：

肿瘤细胞高表达P糖蛋白(P-gp)会抑制化疗的疗效, 导致肿瘤耐药, 而P-gp抑制剂可逆转P-gp介导的肿瘤耐药。辣椒素是辣椒的主要活性成分, 也是P-gp抑制剂。本研究旨在初步探讨辣椒素对多柔比星(DOX)耐药的乳腺癌MCF-7/DOX细胞的耐药逆转作用。细胞毒性研究结果表明, 长期处理72 h后, 辣椒素(50 μM)可以逆转MCF-7/DOX细胞对多柔比星的耐药(逆转倍数为4.68倍), 并浓度依赖性地下调MDR1基因mRNA的表达。在短期培养3 h后, 辣椒素可逆地、浓度依赖性地增加了DOX在MCF-7/DOX细胞中的蓄积, 对DOX在细胞内亚细胞分布的影响不同于阳性对照药维拉帕米。辣椒素也能浓度依赖性地抑制线粒体生成ATP。综上所述, 辣椒素能够逆转MCF-7/DOX细胞对DOX的耐药, 可成为一种很有前途的逆转多药耐药的先导化合物。

关键词: MDR1, 辣椒素, MCF-7/DOX, P糖蛋白

Abstract:

Overexpression of P-glycoprotein (P-gp), encoded by the MDR1 gene, in tumor cells curtails the efficacy of chemotherapy, leading to multidrug resistance (MDR), which can be reversed by the P-gp inhibitor. Capsaicin, the principal pungent component of hot chili, is a reported P-gp inhibitor with anti-cancer properties. In the present study, we aimed to evaluate the MDR reversal effect of capsaicin. The MCF-7/DOX cells were treated with capsaicin for a short term (3 h) or long term (72 h). The cytotoxicity studies were completed by using the SRB method. RT-PCR method was utilized to evaluate the effect of capsaicin on the expression of MDR1 at the mRNA level. The accumulation and subcellular distribution studies were implemented to further investigate the reversal effect of capsaicin. The effect of capsaicin on the ATP production of mitochondria was also evaluated. The results of the cytotoxicity study indicated that capsaicin (50 μM) reversed the resistance of MCF-7/DOX cells to doxorubicin (DOX) with the reversal fold of 4.68, and concentration-dependently down-regulated the expression of MDR1 at the mRNA level after long-term (72 h) incubation. After short-term (3 h) incubation, capsaicin reversibly and concentration-dependently increased the accumulation of DOX into MCF-7/DOX cells and induced a different subcellular distribution of DOX compared with verapamil as a positive control. The ATP production of mitochondria was also concentration-dependently inhibited by capsaicin. In conclusion, capsaicin was capable of reversing the resistance of MCF-7/DOX cells to DOX, making it a promising lead compound for MDR reversal.

Key words: MDR1, Capsaicin, MCF-7/DOX, P-glycoprotein

Supporting:

周毅, 王婷, 赵佳佳, 蒋学华. 辣椒素对多柔比星耐药的乳腺癌细胞的耐药逆转作用[J]. 中国药学(英文版), 2021, 30(8): 666-674.

Yi Zhou, Ting Wang, Jiajia Zhao, Xuehua Jiang. The reversal effect of capsaicin on doxorubicin-resistant breast cancer cells[J]. Journal of Chinese Pharmaceutical Sciences, 2021, 30(8): 666-674.

图/表 8

Figure 1. The chemical structure of capsaicin.

Figure 2. The expression of MDR1 in MCF-7 and MCF-7/DOX cells.

Figure 3. The inhibition curve of DOX in combination with or without capsaicin in MCF-7/DOX cells. Cap, capsaicin; Cap 12.5 + DOX, 12.5 μM of capsaicin combined with DOX; Cap 25 + DOX, 25 μM of capsaicin combined with DOX; Cap 50 + DOX, 50 μM of capsaicin combined with DOX.

Table 1. Cytotoxicity of DOX with or without capsaicin in MCF-7/DOX cells.

Figure 4. The effects of different concentrations of capsaicin on the MDR1 gene expression in MCF-7/DOX cells after 72 h of incubation. Control, untreated MCF-7/DOX cells; Cap, capsaicin; Cap 12.5, 12.5 μM of capsaicin; Cap 25, 25 μM of capsaicin; Cap 50, 50 μM of capsaicin. *P < 0.05.

Figure 5. The accumulation of DOX in MCF-7/DOX cells after 3 h of treatment of different concentrations of a reversal agent. Cap, capsaicin; Ver, verapamil; CDOX/Cprotein, the DOX fluorescence was normalized to the protein concentration. *P < 0.05.

Figure 6. The subcellular accumulation of DOX in MCF-7/DOX cells after 3 h of treatment of a reversal agent. Cap, capsaicin, 150 μM; Ver, verapamil, 150 μM; CDOX/Cprotein, the DOX fluorescence was normalized to the protein concentration. *P < 0.05.

Figure 7. The effects of capsaicin on the ATP production in MCF-7/DOX cells after 3 h (A) or 72 h (B) of incubation. CATP/Cprotein, the ATP concentration was normalized to the protein concentration. Cap, capsaicin; Cap 25, 25 μM of capsaicin; Cap 50, 50 μM of capsaicin; Cap 150, 150 μM of capsaicin; Cap 250, 250 μM of capsaicin. *P < 0.05.

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辣椒素对多柔比星耐药的乳腺癌细胞的耐药逆转作用

The reversal effect of capsaicin on doxorubicin-resistant breast cancer cells

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