Vig Shruti, Srivastava Payal, Rahman Idrisa, Jaranson Renee, Dasgupta Anika, Perttilä Robert, Uusimaa Petteri, Huang Huang-Chiao
Fischell Department of Bioengineering, University of Maryland, College Park, MD 20742, USA.
Authors contributed equally.
Cancer Drug Resist. 2024 Sep 21;7:35. doi: 10.20517/cdr.2024.50. eCollection 2024.
ATP-binding cassette (ABC) transporters are proteins responsible for the efflux of drug molecules from cancer cells, reducing the efficacy of anti-cancer treatments. This study assesses the susceptibility of a panel of clinically used photosensitizers to be transported by ABC transporters The involvement of P-glycoprotein (P-gp/ABCB1), breast cancer resistance protein (BCRP/ABCG2), and multidrug resistance-associated protein 1 (MRP1/ABCC1) in the transport of 7 clinically utilized photosensitizers [benzoporphyrin derivative (BPD), temoporfin, redaporfin, talaporfin sodium, rose bengal, methylene blue, and indocyanine green] were investigated using human breast cancer cell lines following well-established protocols. Briefly, parental MCF-7 cells and sublines that overexpress P-gp (MCF-7 TX400), ABCG2 (MCF-7 MX100), or MRP1 (MCF-7/VP) were treated with photosensitizers with and without ABC transporter inhibitors. Intracellular levels of photosensitizers were measured using extraction method and flow cytometry to determine whether the ABC transporters are associated with efflux or uptake of photosensitizers. The ABCG2 inhibitor (fumitremorgin C) and P-gp inhibitor (valspodar) effectively blocked the transport mediated by ABCG2 and P-gp of rose bengal and BPD. Redaporfin showed increased accumulation in the presence of valspodar with flow cytometry. Interestingly, MCF-7/VP cells were found to have reduced intracellular accumulation of rose bengal, which was restored with MRP1 inhibitor (MK571). The cell viability assay showed photodynamic therapy (PDT) resistance with Redaporfin in P-gp-overexpressing cells, BPD in ABCG2- and P-gp-overexpressing cells, and with Rose bengal in ABCG2-, P-gp- and MRP1-overexpressing cells, respectively. However, no change in intracellular retention was observed for other photosensitizers. In summary, our study provided new knowledge that temoporfin, talaporfin sodium, methylene blue, and indocyanine green are not substrates of ABCG2, P-gp, or MRP1. Redaporfin is a substrate for P-gp. BPD is a known substrate of ABCG2 and P-gp. Rose bengal is a substrate of ABCG2, P-gp, and MRP1. The results presented here indicate ABC transporter substrate status as a possible cause for cellular resistance to photodynamic therapy with rose bengal, redaporfin, and BPD.
ATP结合盒（ABC）转运蛋白是负责将药物分子从癌细胞中排出的蛋白质，这会降低抗癌治疗的效果。本研究评估了一组临床使用的光敏剂被ABC转运蛋白转运的敏感性。采用成熟的方案，利用人乳腺癌细胞系研究了P-糖蛋白（P-gp/ABCB1）、乳腺癌耐药蛋白（BCRP/ABCG2）和多药耐药相关蛋白1（MRP1/ABCC1）在7种临床使用的光敏剂[苯并卟啉衍生物（BPD）、替莫泊芬、瑞波芬、他拉泊芬钠、孟加拉玫瑰红、亚甲蓝和吲哚菁绿]转运中的作用。简要地说，用或不用ABC转运蛋白抑制剂处理亲本MCF-7细胞以及过表达P-gp（MCF-7 TX400）、ABCG2（MCF-7 MX100）或MRP1（MCF-7/VP）的亚系细胞。使用萃取法和流式细胞术测量光敏剂的细胞内水平，以确定ABC转运蛋白是否与光敏剂的外排或摄取有关。ABCG2抑制剂（烟曲霉毒素C）和P-gp抑制剂（伐司朴达）有效地阻断了孟加拉玫瑰红和BPD由ABCG2和P-gp介导的转运。瑞波芬在伐司朴达存在的情况下，通过流式细胞术显示其积累增加。有趣的是，发现MCF-7/VP细胞内孟加拉玫瑰红的积累减少，而用MRP1抑制剂（MK571）可使其恢复。细胞活力测定表明，在过表达P-gp的细胞中，瑞波芬对光动力疗法（PDT）耐药；在过表达ABCG2和P-gp的细胞中，BPD对PDT耐药；在过表达ABCG2、P-gp和MRP1的细胞中，孟加拉玫瑰红对PDT耐药。然而，未观察到其他光敏剂的细胞内滞留发生变化。总之，我们的研究提供了新的知识，即替莫泊芬、他拉泊芬钠、亚甲蓝和吲哚菁绿不是ABCG2、P-gp或MRP1的底物。瑞波芬是P-gp的底物。BPD是ABCG2和P-gp的已知底物。孟加拉玫瑰红是ABCG2、P-gp和MRP1的底物。此处呈现的结果表明ABC转运蛋白底物状态可能是细胞对孟加拉玫瑰红、瑞波芬和BPD光动力疗法耐药的原因。
