黄连碱通过抑制ABC转运蛋白的功能和表达来增强化疗耐药乳腺癌细胞的敏感性。

Coptisine enhances the sensitivity of chemoresistant breast cancer cells by inhibiting the function and expression of ABC transporters.

作者信息

Eid Safaa Yehia

机构信息

Department of Biochemistry, Faculty of Medicine, Umm Al-Qura University, Makkah, Saudi Arabia.

出版信息

Front Pharmacol. 2024 Dec 3;15:1472458. doi: 10.3389/fphar.2024.1472458. eCollection 2024.

DOI:10.3389/fphar.2024.1472458

PMID:39691399

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC11650208/

Abstract

BACKGROUND

Multidrug resistance (MDR), mainly caused by ATP-binding cassette transporters (ABCTs) efflux, makes it difficult for many anticancer drugs to treat breast cancer (BC). Phytochemicals can reverse cancer's MDR by modifying ABC transporter expression and function, as well as working synergistically with anticancer drugs to target other molecules. The reversal effect of the isoquinoline alkaloid coptisine (COP) was assessed on four breast cell lines; Two sensitive MCF-7 cell lines with positive estrogen, androgen, progesterone, and glucocorticoid receptors, as well as MDB-MB-231 cells with negative estrogen, progesterone, and HER2 receptors, and two doxorubicin-resistant cell lines, MCF-7/ADR and MDB-MB-231/ADR.

METHODS

The cytotoxicity of COP and its ability to improve doxorubicin (DOX) cytotoxicity were assessed using the MTT assay. The effectiveness of COP in reversing DOX resistance was evaluated by calculating resistance ratio (RR) values, combination index (CI), and isobologram (IB). The inhibitory effect of COP on ABCT efflux function in comparison to verapamil (VER) was evaluated by measuring the cellular accumulation of Rho123 using flow cytometry. The impact of COP, either alone or in combination with DOX, on the gene expression of ABCTs (P-gp/MDR1, BCRP, and MRP1) of investigated cell lines was assessed by RT-PCR.

RESULTS

The COP showed modest cytotoxicity on the examined cell lines. In MCF-7/ADR and MDA-MB-231/ADR cells, COP (31 μM) enhanced DOX cytotoxicity with CI (0.77 and 0.75), RR (2.58 and 3.33), and IB suggesting synergism. COP significantly inhibits ABCT function in resistant BC cell lines, increases Rho123 accumulation, and decreases efflux more than VER; 2.1 and 1.2-fold, respectively. The combination of COP and DOX had a strong inhibitory effect on ABCT function (3.1 and 3.9 times VER, P< 0.001) and downregulated the genes and protein expression of ABCT.

CONCLUSION

COP reversed ABCT-mediated multidrug resistance , indicating its potential as a multidrug resistance-reversing agent in cancer chemotherapy.

摘要

背景

多药耐药性（MDR）主要由ATP结合盒转运蛋白（ABCTs）外排引起，这使得许多抗癌药物难以治疗乳腺癌（BC）。植物化学物质可通过改变ABC转运蛋白的表达和功能来逆转癌症的MDR，还可与抗癌药物协同作用以靶向其他分子。评估了异喹啉生物碱黄连碱（COP）对四种乳腺癌细胞系的逆转作用；两种雌激素、雄激素、孕激素和糖皮质激素受体呈阳性的敏感MCF-7细胞系，以及雌激素、孕激素和HER2受体呈阴性的MDB-MB-231细胞系，还有两种对多柔比星耐药的细胞系，MCF-7/ADR和MDB-MB-231/ADR。

方法

使用MTT法评估COP的细胞毒性及其提高多柔比星（DOX）细胞毒性的能力。通过计算耐药比（RR）值、联合指数（CI）和等效线图（IB）来评估COP逆转DOX耐药性的有效性。通过流式细胞术测量Rho123的细胞内蓄积，评估COP与维拉帕米（VER）相比对ABCT外排功能的抑制作用。通过RT-PCR评估COP单独或与DOX联合对所研究细胞系ABCTs（P-糖蛋白/MDR1、BCRP和MRP1）基因表达的影响。

结果

COP在所检测的细胞系中显示出适度的细胞毒性。在MCF-7/ADR和MDA-MB-231/ADR细胞中，COP（31μM）增强了DOX的细胞毒性，CI分别为0.77和0.75，RR分别为2.58和3.33，等效线图表明具有协同作用。COP显著抑制耐药乳腺癌细胞系中的ABCT功能，增加Rho123蓄积，且比VER更能降低外排；分别为VER的2.1倍和1.2倍。COP与DOX联合对ABCT功能有强烈抑制作用（是VER的3.1倍和3.9倍，P<0.001），并下调ABCT的基因和蛋白表达。