Hosseinkhani Negar, Alipour Shiva, Ghaffari Jolfayi Amir, Aghebati-Maleki Leili, Baghbani Elham, Alizadeh Nazila, Khaze Vahid, Baradaran Behzad
Immunology Research Center, Tabriz University of Medical Sciences, Tabriz, Iran.
Department of Immunology, Faculty of Medicine, Tabriz University of Medical Sciences, Tabriz, Iran.
Cytotechnology. 2025 Feb;77(1):19. doi: 10.1007/s10616-024-00677-4. Epub 2024 Dec 12.
Breast cancer is the most frequent cancer in women with a 20% mortality rate. The fate of patients suffering from breast cancer can be influenced by immune cells and tumor cells interaction in the tumor microenvironment (TME). Immune checkpoints such as Cytotoxic T-lymphocyte-associated antigen-4 (CTLA-4) are regulators of the immune system and defend normal tissues from immune cell attacks but they can be expressed in breast cancer tissue and facilitate immune evasion of tumoral cells. Based on this, here we studied the role of CTLA-4 silencing by specific siRNA in MCF-7 breast cancer cell line together with Docetaxel treatment which is one of the robust chemotherapy agents to demonstrate the significance of combining chemotherapy with efficient targeted therapy in tumor regression. The MCF-7 breast cancer cell line was transfected with CTLA-4-siRNA through the electroporation method, then received an appropriate dose of Docetaxel determined by MTT assay. Flow cytometry was utilized to investigate the consequence of simultaneous CTLA-4 gene silencing and Docetaxel treatment on the apoptosis and cell cycle of MCF-7 cells. The expression levels of Bax and Bcl-2 were also investigated using quantitative real-time PCR. Compared to control groups, CTLA-4-suppressed and Docetaxel-treated cells became more susceptible to apoptosis and cell cycle arrest at the G2-M phase. The additive effect of CTLA-4 knockdown together with Docetaxel treatment significantly downregulated BCL-2 level and upregulated BAX expression. Our findings support the idea that combining chemotherapy such as Docetaxel with efficient targeted therapy against inhibitory immune checkpoints can be a promising strategy in cancer treatment.
乳腺癌是女性中最常见的癌症,死亡率为20%。乳腺癌患者的命运会受到肿瘤微环境(TME)中免疫细胞与肿瘤细胞相互作用的影响。免疫检查点,如细胞毒性T淋巴细胞相关抗原4(CTLA-4),是免疫系统的调节因子,可保护正常组织免受免疫细胞攻击,但它们可在乳腺癌组织中表达,促进肿瘤细胞的免疫逃逸。基于此,我们在此研究了通过特异性小干扰RNA(siRNA)沉默CTLA-4在MCF-7乳腺癌细胞系中的作用,并联合多西他赛治疗(多西他赛是一种强效化疗药物),以证明化疗与有效的靶向治疗相结合在肿瘤消退中的重要性。通过电穿孔法用CTLA-4-siRNA转染MCF-7乳腺癌细胞系,然后通过MTT试验确定合适剂量的多西他赛。利用流式细胞术研究同时沉默CTLA-4基因和多西他赛治疗对MCF-7细胞凋亡和细胞周期的影响。还使用定量实时PCR研究Bax和Bcl-2的表达水平。与对照组相比,CTLA-4抑制和多西他赛处理的细胞在G2-M期更容易发生凋亡和细胞周期阻滞。CTLA-4基因敲低与多西他赛治疗的相加效应显著下调BCL-2水平并上调BAX表达。我们的研究结果支持这样一种观点,即多西他赛等化疗与针对抑制性免疫检查点的有效靶向治疗相结合可能是癌症治疗中一种有前景的策略。