目的:尽管慢性粒细胞白血病(CML)的治疗方法有了很大的改进,由于耐药性,这种恶性肿瘤被认为是无法治愈的。然而,研究CML的分子机制可能会导致开发非常有效的靶向治疗方法,从而改善患者的预后。碱性亮氨酸拉链转录因子ATF-like3(BATF3),作为转录因子,被认为是细胞活性的关键调节剂,其功能已在几种癌症类型的肿瘤发展和生长中得到评估。本研究旨在评估siRNA介导的BATF3下调通过细胞增殖对CML癌细胞的细胞影响的潜力。诱导凋亡,和细胞周期分布。
方法:通过电穿孔装置将BATF3siRNA转染到K562CML细胞中。为了测量细胞活力和细胞凋亡,MTT法和膜联蛋白V/PI染色,分别。此外,应用细胞周期检测和流式细胞仪检测K562细胞的细胞周期分布。有关更多验证,通过定量实时聚合酶链反应(qRT-PCR)相对评估相关基因的mRNA表达。
结果:数据表明,siRNA介导的BATF3失活严重促进了细胞凋亡。此外,靶向治疗导致Caspase-3基因高表达和Bax/Bcl-2比值。与对照相比,沉默的BATF3还在sub-G1期诱导细胞周期停滞。最后,通过抑制CML细胞中的BATF3,c-Myc基因表达显着下降。
结论:这项研究的结果表明,抑制BATF3是CML的有效靶向治疗策略。
OBJECTIVE: Despite considerable improvement in therapeutic approaches to chronic myeloid leukemia (CML) treatment, this malignancy is considered incurable due to resistance. However, investigating the molecular mechanism of CML may give rise to the development of extremely efficient targeted therapies that improve the prognosis of patients. Basic leucine zipper transcription factor ATF-like3 (
BATF3), as transcription factor, is considered a key regulator of cellular activities and its function has been evaluated in tumor development and growth in several cancer types. This study aimed to evaluate the potential of the cellular impact of siRNA-mediated downregulation of
BATF3 on CML cancer cells through cell proliferation, induction of apoptosis, and cell cycle distribution.
METHODS: The transfection of
BATF3 siRNA to K562 CML cells was performed by electroporation device. To measure cellular viability and apoptosis, MTT assay and Annexin V/PI staining were carried out, respectively. Also, cell cycle assay and flow cytometry instrument were applied to assess cell cycle distribution of K562 cells. For more validation, mRNA expression of correlated genes was relatively evaluated by quantitative real-time polymerase chain reaction (qRT-PCR).
RESULTS: The data indicated that siRNA-mediated BATF3 inactivating severely promoted the cell apoptosis. Also, the targeted therapy led to high expression of Caspase-3 gene and Bax/Bcl-2 ratio. Silenced
BATF3 also induced cell cycle arrest in phase sub-G1 compared to control. Finally, a noticeable decrement was obtained in c-Myc gene expression through suppression of
BATF3 in CML cells.
CONCLUSIONS: The findings of this research illustrated the suppression of BATF3 as an effective targeted therapy strategy for CML.