Khordadmehr Monireh, Matin Reyhaneh, Baradaran Behzad, Baghbani Elham, Jigari-Asl Farinaz, Noorolyai Saeed
Department of Pathobiology, Faculty of Veterinary Medicine, University of Tabriz, 51665-1647, Tabriz, Iran.
Immunology Research Center, Tabriz University of Medical Sciences, 51666-14761, Tabriz, Iran.
Adv Pharm Bull. 2023 Mar;13(2):378-384. doi: 10.34172/apb.2023.041. Epub 2022 Jan 5.
MicroRNAs (miRNAs) can contribute to cancer initiation, development, and progression. In this study, the effect of miRNA-4800 restoration on the growth and migration inhibition of human breast cancer (BC) cells was investigated. For this purpose, transfection of miR-4800 was performed into MDA-MB-231 BC cells using jetPEI. Subsequently, the expression levels of miR-4800 and CXCR4, ROCK1, CD44, and vimentin genes were measured using quantitative real-time polymerase chain reaction (q-RT-PCR) and specific primers. Also, the proliferation inhibition and apoptosis induction of cancer cells were evaluated by MTT and flow cytometry (Annexin V-PI method) techniques, respectively. Additionally, cancer cell migration after miR-4800 transfection was assessed by wound-healing (scratch) assay. The restoration of miR-4800 in MDA-MB-231 cells resulted in the decreased expression level of CXCR4 ( ˂ 0.01), ROCK1 ( ˂ 0.0001), CD44 ( ˂ 0.0001), and vimentin ( ˂ 0.0001) genes. Also, MTT results showed restoration of miR-4800 could significantly reduce cell viability rate ( ˂ 0.0001) compared with the control group. Cell migration remarkably inhibited ( ˂ 0.001) upon miR-4800 transfection in treated BC cells. Flow cytometry data demonstrated that miR-4800 replacement considerably induced apoptosis in cancer cells ( ˂ 0.001) compared with control cells. Taken together, it seems that miR-4800 can act as a tumor suppressor miRNA in BC and play an essential role in modulating apoptosis, migration, and metastasis in BC. Therefore, it may be suggested as a potential therapeutic target in treating BC by performing additional tests in the future.
微小RNA（miRNA）可促进癌症的起始、发展和进展。在本研究中，研究了miRNA - 4800恢复对人乳腺癌（BC）细胞生长和迁移抑制的影响。为此，使用jetPEI将miR - 4800转染到MDA - MB - 231 BC细胞中。随后，使用定量实时聚合酶链反应（q - RT - PCR）和特异性引物测量miR - 4800以及CXCR4、ROCK1、CD44和波形蛋白基因的表达水平。此外，分别通过MTT和流式细胞术（膜联蛋白V - PI法）技术评估癌细胞的增殖抑制和凋亡诱导情况。另外，通过伤口愈合（划痕）试验评估miR - 4800转染后癌细胞的迁移情况。MDA - MB - 231细胞中miR - 4800的恢复导致CXCR4（˂0.01）、ROCK1（˂0.0001）、CD44（˂0.0001）和波形蛋白（˂0.0001）基因的表达水平降低。此外，MTT结果显示，与对照组相比，miR - 4800的恢复可显著降低细胞活力率（˂0.0001）。在处理的BC细胞中，miR - 4800转染后细胞迁移受到显著抑制（˂0.001）。流式细胞术数据表明，与对照细胞相比，miR - 4800的替代显著诱导癌细胞凋亡（˂0.001）。综上所述，miR - 4800似乎可作为BC中的一种肿瘤抑制性miRNA，并在调节BC的凋亡、迁移和转移中发挥重要作用。因此，未来通过进行更多测试，它可能被建议作为治疗BC的潜在治疗靶点。
