Background: Chronic myeloid leukemia (CML) is considered a type of hematopoietic stem cell disease that affects the bone marrow and blood. Objective: This study aimed to investigate the possible role of the Periplaneta americana extract CII-3 (PAE CII-3) in the aging of K562 cells. Materials and Methods: The proliferation and cell cycle of K562 cells were determined using the CCK-8 assay and the cell cycle assay, respectively. K562 cells were stained with SA-β-Gal to evaluate cell aging. The mitochondrial membrane potential of K562 cells was detected with the JC-1 mitochondrial membrane potential assay kit. Telomerase activity was verified using the PCR assay. The transcription of silencing information regulator 2 related enzyme 1 (SIRT1), TSC2, and the mTOR gene were evaluated with RT-PCR assay. The expression of SIRT1, p-TSC2, and p-mTOR was examined using a Western blot assay. Results: PAE CII-3 at all concentrations (5, 10, 20, 40, 80, 160 µg/mL) demonstrated obvious inhibitory effects on K562 cell proliferation, among which 80 µg/mL showed the highest inhibitory effect. PAE CII-3 significantly blocked the cell cycle and reduced the colony-forming unit (CFU) of K562 cells compared to those in the Control group (p < 0.001). PAE CII-3 markedly increased positive SA-β-Gal staining K562 cells compared to the Control group (p < 0.001). PAE CII-3 significantly reduced mitochondrial membrane potential and decreased TERT gene transcription in K562 cells compared to those of the Control group (p < 0.001). The transcription of the SIRT1 gene (p < 0.01) and the TCS2 gene (p < 0.001) was markedly decreased, and the transcription of the mTOR gene (p < 0.05) was significantly increased in K562 cells treated with PAE CII-3 compared to those of the Control group. PAE CII-3 significantly decreased the expression of SIRT1 (p < 0.01) and p-TSC2 (p < 0.001) and upregulated the expression of p-mTOR (p < 0.01) in K562 cells compared to those of the Control group. Conclusion: PAE CII-3 treatment could trigger aging in K562 cells by activating the SIRT1/TSC2/mTOR signaling pathway. This study would provide a potential hypothesis of the mechanism by which PAE CII-3 treatment induces the aging of chronic myeloid leukemia cells.

中文翻译：

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美洲大蠊提取物 CII-3 (PAE CII-3) 通过调节 SIRT1/TSC2/mTOR 分子触发 K562 细胞衰老

背景：慢性粒细胞白血病（CML）被认为是一种影响骨髓和血液的造血干细胞疾病。目的：本研究旨在探讨美洲大蠊提取物CII-3（PAE CII-3）在K562细胞衰老中的可能作用。材料和方法：分别使用CCK-8测定和细胞周期测定测定K562细胞的增殖和细胞周期。K562 细胞用 SA-β-Gal 染色以评估细胞老化。采用JC-1线粒体膜电位测定试剂盒检测K562细胞线粒体膜电位。使用 PCR 测定验证端粒酶活性。通过 RT-PCR 测定评估沉默信息调节因子 2 相关酶 1 (SIRT1)、TSC2 和 mTOR 基因的转录。使用蛋白质印迹法检查 SIRT1、p-TSC2 和 p-mTOR 的表达。结果：PAE CII-3各浓度（5、10、20、40、80、160 μg/mL）对K562细胞增殖均有明显的抑制作用，其中80 μg/mL抑制作用最强。与对照组相比，PAE CII-3 显着阻断细胞周期并减少 K562 细胞的集落形成单位 (CFU) (p < 0.001)。与对照组相比，PAE CII-3 显着增加 SA-β-Gal 染色阳性 K562 细胞 (p < 0.001)。与对照组相比，PAE CII-3 显着降低 K562 细胞中的线粒体膜电位并减少 TERT 基因转录（p < 0.001）。与 PAE CII-3 处理的 K562 细胞相比，SIRT1 基因 (p < 0.01) 和 TCS2 基因 (p < 0.001) 的转录显着降低，mTOR 基因的转录 (p < 0.05) 显着增加。对照组的那些。与对照组相比，PAE CII-3 显着降低 K562 细胞中 SIRT1 (p < 0.01) 和 p-TSC2 (p < 0.001) 的表达，并上调 p-mTOR 的表达 (p < 0.01)。结论：PAE CII-3治疗可通过激活SIRT1/TSC2/mTOR信号通路引发K562细胞衰老。这项研究将为 PAE CII-3 治疗诱导慢性粒细胞白血病细胞衰老的机制提供一个潜在的假设。