Department of Animal Biology, Faculty of Natural Sciences, University of Tabriz, Tabriz, Iran.
Immunology Research Center, Tabriz University of Medical Sciences, Tabriz, Iran.
Mol Biol Rep. 2023 Oct;50(10):8407-8420. doi: 10.1007/s11033-023-08711-y. Epub 2023 Aug 24.
Chemotherapy is a predominant strategy for breast cancer (BC) treatment and paclitaxel (PTX) has been known as a conventional chemotherapeutic drug. However, insensitivity of BC cells to PTX limits the anti-tumor effects of this agent. MicroRNAs are closely related to BC which are suggested as therapeutic factors in the combination therapy of BC. We examined the possible efficacy of miR-138-5p restoration in combination with PTX to impove BC treatment.
The human breast cancer cell line MDA-MB-231 was transfected with miR-138-5p mimics and treated with PTX, in a combined or separate manner. The MTT assay was accomplished to determine inhibitory doses of PTX. Annexin V/PI assay and DAPI staining were applied to evaluate apoptosis. Flow cytometry was applied to determine cells arrested in different phases of the cell-cycle. Expression levels of molecular factors involved in cell migration, proliferation, apoptosis, and cell cycle were determined via western blotting and qRT-PCR.
MiR-138-5p combined with PTX suppressed cell migration via modulating MMP2, E-cadherin, and vimentin and sustained colony formation and proliferation by downregulation of the PI3K/AKT pathway. qRT-PCR showed that miR-138-5p increases BC chemosensitivity to PTX by regulating the apoptosis factors, including Bcl-2, Bax, Caspase 3, and Caspase 9. Moreover, miR-138-5p restoration and paclitaxel therapy combined arrest the cells in the sub-G and G phases of cell cycle by regulating p21, CCND1, and CDK4.
Restored miR-138-5p intensified the chemosensitivity of MDA-MB-231 cell line to PTX, and the combination of miR-138-5p with PTX might represent a novel approach in BC treatment.
化疗是乳腺癌（BC）治疗的主要策略，紫杉醇（PTX）已被视为常规化疗药物。然而，BC 细胞对 PTX 的不敏感性限制了该药物的抗肿瘤作用。microRNAs 与 BC 密切相关，被认为是 BC 联合治疗的治疗因素。我们研究了 miR-138-5p 恢复与 PTX 联合治疗以改善 BC 治疗的可能效果。
人乳腺癌细胞系 MDA-MB-231 用 miR-138-5p 模拟物转染，并以联合或单独方式用 PTX 处理。MTT 测定法确定 PTX 的抑制剂量。Annexin V/PI 测定法和 DAPI 染色用于评估细胞凋亡。流式细胞术用于确定细胞在细胞周期的不同阶段停滞。通过 Western blot 和 qRT-PCR 测定参与细胞迁移、增殖、凋亡和细胞周期的分子因子的表达水平。
miR-138-5p 与 PTX 联合通过调节 MMP2、E-钙粘蛋白和波形蛋白抑制细胞迁移，并通过下调 PI3K/AKT 通路维持集落形成和增殖。qRT-PCR 显示 miR-138-5p 通过调节凋亡因子，包括 Bcl-2、Bax、Caspase 3 和 Caspase 9，增加 BC 对 PTX 的化疗敏感性。此外，miR-138-5p 恢复和紫杉醇治疗联合通过调节 p21、CCND1 和 CDK4 将细胞周期阻滞在 sub-G 和 G 期。
恢复的 miR-138-5p 增强了 MDA-MB-231 细胞系对 PTX 的化疗敏感性，miR-138-5p 与 PTX 的联合可能代表 BC 治疗的一种新方法。
