Rajabi Neda, Mohammadnejad Fateme, Doustvandi Mohammad Amin, Shadbad Mahdi Abdoli, Amini Mohammad, Tajalli Habib, Mokhtarzadeh Ahad, Baghbani Elham, Silvestris Nicola, Baradaran Behzad
Immunology Research Center, Tabriz University of Medical Sciences, Tabriz, Iran.
Immunology Research Center, Tabriz University of Medical Sciences, Tabriz, Iran; Student Research Committee, Tabriz University of Medical Sciences, Tabriz, Iran.
Photodiagnosis Photodyn Ther. 2023 Mar;41:103212. doi: 10.1016/j.pdpdt.2022.103212. Epub 2022 Nov 25.
Photodynamic therapy (PDT) is a light-based anti-neoplastic therapeutic approach. Growing evidence indicates that combining conventional anti-cancer therapies with PDT can be a promising approach to treat malignancies. Herein, we aimed to investigate anti-cancer effects of the combination treatment of zinc phthalocyanine (ZnPc)-PDT with tamoxifen (TA) on MDA-MB-231 cells (as a triple-negative breast cancer (TNBC) cell line). For this purpose, we investigated the cytotoxicity of TA and ZnPc-PDT on MDA-MB-231 cells performing the MTT assay. The effect of TA and ZnPc-PDT on the apoptosis of MDA-MB-231 cells was studied using Annexin V/PI and DAPI staining. The wound-healing assay, and colony formation assay were performed to study the effect of TA and ZnPc-PDT on the migration, and clonogenicity of MDA-MB-231 cells, respectively. The qRT-PCR was done to study the gene expression of caspase-8, caspase-9, caspase-3, ZEB1, ROCK1, SNAIL1, CD133, CD44, SOX2, and ABCG2 (ATP-binding cassette sub-family G member 2). Based on our results, monotherapies with TA and ZnPc-PDT can remarkably increase cell cytotoxicity effects, stimulate apoptosis via downregulating Bcl-2 and upregulating caspase-3 and caspase-9, inhibit migration via downregulating SNAIL1 and ZEB1, and suppress clonogenicity via downregulating SOX2 and CD44 in MDA-MB-231 cells. Besides, these monotherapies can downregulate the expression of ABCG2 in MDA-MB-231 cells. Nevertheless, the combination treatment can potentiate the above-mentioned anti-cancer effects compared to monotherapy with TA. Of interest, the combined treatment of TA with ZnPc-PDT can synergically increase cell cytotoxicity effects on MDA-MB-231 cells. In fact, synergistic effects were estimated by calculation of Combination Index (CI); that synergistic outcomes were observed in all groups. Also, this combination treatment can significantly upregulate the caspase-8 gene expression and downregulate ROCK1 and CD133 gene expression in MDA-MB-231 cells. Overall, our results show that ZnPc-PDT can more sensitize the MDA-MB-231 cells to TA treatment. Based on our knowledge and experiment, the synergistic effects of ZnPc-PDT and TA deserve further evaluation in cancer research.
光动力疗法（PDT）是一种基于光的抗肿瘤治疗方法。越来越多的证据表明，将传统抗癌疗法与光动力疗法相结合可能是治疗恶性肿瘤的一种有前景的方法。在此，我们旨在研究锌酞菁（ZnPc）-光动力疗法与他莫昔芬（TA）联合治疗对MDA-MB-231细胞（作为三阴性乳腺癌（TNBC）细胞系）的抗癌作用。为此，我们通过MTT试验研究了TA和ZnPc-光动力疗法对MDA-MB-231细胞的细胞毒性。使用膜联蛋白V/碘化丙啶和4',6-二脒基-2-苯基吲哚染色研究了TA和ZnPc-光动力疗法对MDA-MB-231细胞凋亡的影响。分别进行伤口愈合试验和集落形成试验，以研究TA和ZnPc-光动力疗法对MDA-MB-231细胞迁移和克隆形成能力的影响。进行qRT-PCR以研究半胱天冬酶-8、半胱天冬酶-9、半胱天冬酶-3、锌指E盒结合蛋白1（ZEB1）、Rho相关卷曲螺旋蛋白激酶1（ROCK1）、锌指转录抑制因子1（SNAIL1）、CD133、CD44、性别决定区Y框蛋白2（SOX2）和ATP结合盒亚家族G成员2（ABCG2）的基因表达。基于我们的结果，TA和ZnPc-光动力疗法单一疗法可显著增加细胞毒性作用，通过下调Bcl-2和上调半胱天冬酶-3和半胱天冬酶-9来刺激凋亡，通过下调SNAIL1和ZEB1来抑制迁移，并通过下调SOX2和CD44来抑制MDA-MB-231细胞的克隆形成能力。此外，这些单一疗法可下调MDA-MB-231细胞中ABCG2的表达。然而，与TA单一疗法相比，联合治疗可增强上述抗癌作用。有趣的是，TA与ZnPc-光动力疗法联合治疗可协同增加对MDA-MB-231细胞的细胞毒性作用。事实上，通过计算联合指数（CI）来评估协同效应；在所有组中均观察到协同结果。此外，这种联合治疗可显著上调MDA-MB-231细胞中半胱天冬酶-8基因表达并下调ROCK1和CD133基因表达。总体而言，我们的结果表明ZnPc-光动力疗法可使MDA-MB-231细胞对TA治疗更敏感。基于我们的知识和实验，ZnPc-光动力疗法和TA的协同效应在癌症研究中值得进一步评估。
