Öner Çağrı, Köser Faruk, Çolak Ertuğrul
Department of Medical Biology, Kırklareli University, Faculty of Medicine, Kırklareli, Turkey.
Department of Medical and Genetics, Maltepe University, Faculty of Medicine, İstanbul, Turkey.
Nucleosides Nucleotides Nucleic Acids. 2024 Dec 4:1-17. doi: 10.1080/15257770.2024.2437037.
PIWI-Interacting RNAs are small non-coding RNAs derived from single-stranded RNAs which plays a crucial role in epigenetic regulation through transposon silencing and mRNA degradation deamination. This study aimed to inhibit piR-651 and piR-823 in MDA-MB-231 triple-negative breast cancer cells and to explore their potential effects on healthy HUVEC cells. Non-target, anti-piR-651, and anti-piR-823 sequences were transfected in bothHUVEC and MDA-MB-231 cells using Lipofectamine. Proliferation and motility were assessed at 24, 48, and 72 h post-transfection in both cell lines. Based on the motility findings, MDA-MB-231 cells were underwent an invasion assay using crystal violet staining. The expressions of Ki-67, HIF-1α, MMP-2, and MMP-9 genes were measured at 48 h, when both cell lines exhibited the most significant effects of inhibition. The optimal time for proliferation of anti-piR-651 and anti-piR-823 transfected MDA-MB-231 cells was determined to be at 48 h, as indicated by decreased motility and invasion assay results ( < 0.001). NeverthelessHowever, there was no significant difference in the motility and proliferation of HUVECss transfected with anti-piR-651 and anti-piR-823 compared to the control group ( > 0.05). Asides from MMP-2 in anti-piR-823 transfected HUVECs and HIF-1α in anti-piR-823 transfected MDA-MB-231 cells, gene expressions of Ki-67, HIF-1α, MMP-2, and MMP-9 were reduced in both cell lines ( < 0.001). Inhibition of piR-651 and piR-823 decreased the survival and metastasis of cancer cells, without causing vital structural changes in healthy cells. Future research in cancer gene therapy or genetic modification may benefit from investigating piR-651 and piR-823 as possible inhibitors of breast cancer invasion and metastasis.
PIWI相互作用RNA是一类源自单链RNA的小非编码RNA，其通过转座子沉默和mRNA降解脱氨作用在表观遗传调控中发挥关键作用。本研究旨在抑制MDA-MB-231三阴性乳腺癌细胞中的piR-651和piR-823，并探讨它们对健康人脐静脉内皮细胞（HUVEC）的潜在影响。使用脂质体转染试剂将非靶向序列、抗piR-651序列和抗piR-823序列转染至HUVEC和MDA-MB-231细胞中。在转染后24、48和72小时评估两种细胞系的增殖和迁移能力。基于迁移能力的研究结果，使用结晶紫染色法对MDA-MB-231细胞进行侵袭实验。在48小时时检测Ki-67、HIF-1α、MMP-2和MMP-9基因的表达，此时两种细胞系均表现出最显著的抑制效果。抗piR-651和抗piR-823转染的MDA-MB-231细胞增殖的最佳时间确定为48小时，迁移和侵袭实验结果显示其迁移能力下降（<0.001）。然而，与对照组相比，抗piR-651和抗piR-823转染的HUVEC细胞在迁移和增殖方面没有显著差异（>0.05）。除了抗piR-823转染的HUVEC细胞中的MMP-2和抗piR-823转染的MDA-MB-231细胞中的HIF-1α外，两种细胞系中Ki-67、HIF-1α、MMP-2和MMP-9的基因表达均降低（<0.001）。抑制piR-651和piR-823可降低癌细胞的存活和转移能力，且不会对健康细胞造成重大结构改变。癌症基因治疗或基因改造的未来研究可能会受益于将piR-651和piR-823作为乳腺癌侵袭和转移的潜在抑制剂进行研究。
