Shiri Musarreza, Sharifi Mohammadreza, Dianat-Moghadam Hasan, Mehrzad Valiollah
Department of Genetics and Molecular Biology, School of Medicine, Isfahan University of Medical Sciences, Isfahan, 81744-174, Iran.
Department of Internal Medicine, Division of Hematology, School of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran.
Med Oncol. 2025 Sep 15;42(11):474. doi: 10.1007/s12032-025-03015-6.
Acute myeloid leukemia (AML) is a severe hematologic malignancy caused by the rapid proliferation of myeloid progenitor cells in the bone marrow. Non-coding RNAs, particularly, PIWI-interacting RNAs (piRNAs), play a role in the development of several cancers. Hsa-piR-33195, in particular, may promote AML development since its levels are substantially higher in AML patients. This study explores its anticancer effects and molecular mechanisms in AML cells in vitro. Human AML bone marrow blast cells and KG1 cells were transfected with piR-33195 locked nucleic acid antisense (LNA) GapmeRs 24, 48, and 72 hours after transfection. The transfection efficiency was then assessed using a fluorescent microscope. After that, qRT-PCR was used to measure piR-33195, CASP3, and CASP9. MTT was employed to evaluate the growth and viability of treated cells. Finally, the therapeutic effect of the antisense LNA GapmeRs on cell apoptosis and necrosis was assessed using Annexin V/PI staining. Bioinformatics analyses were conducted to predict the piR-33195 target genes in AML. Findings demonstrated a significant decrease in piR-33195, as well as increased CASP3 and CASP9 expression, in bone marrow samples from AML patients and the KG1 cell line, 24, 48, and 72 hours after transfection with antisense LNA Gamers. Moreover, suppressing piR-33195 markedly reduced cell growth in the transfected cells. Apoptosis and necrosis were significantly elevated in these cells. Bioinformatics identified gene and pathways associated with apoptosis and PI3K-AKT signaling. Inhibiting piR-33195 may be a promising treatment option for AML with fewer adverse effects.
急性髓系白血病（AML）是一种严重的血液系统恶性肿瘤，由骨髓中髓系祖细胞的快速增殖引起。非编码RNA，特别是PIWI相互作用RNA（piRNA），在几种癌症的发生发展中发挥作用。特别是hsa-piR-33195，可能促进AML的发展，因为其在AML患者中的水平显著更高。本研究在体外探索其对AML细胞的抗癌作用及分子机制。人AML骨髓原始细胞和KG1细胞在转染24、48和72小时后用piR-33195锁核酸反义（LNA）GapmeRs进行转染。然后使用荧光显微镜评估转染效率。之后，用qRT-PCR检测piR-33195、CASP3和CASP9。采用MTT法评估处理后细胞的生长和活力。最后，使用Annexin V/PI染色评估反义LNA GapmeRs对细胞凋亡和坏死的治疗效果。进行生物信息学分析以预测AML中piR-33195的靶基因。研究结果表明，在转染反义LNA GapmeRs 24、48和72小时后，AML患者骨髓样本和KG1细胞系中piR-33195显著降低，同时CASP3和CASP9表达增加。此外，抑制piR-33195显著降低了转染细胞的生长。这些细胞中的凋亡和坏死显著增加。生物信息学确定了与凋亡和PI3K-AKT信号通路相关的基因和途径。抑制piR-33195可能是一种有前景的AML治疗选择，且副作用较少。
