Department of Radiology, Affiliated Hospital of Dali University, China.
Department of Histology and Embryology, Dali University, China.
Adv Clin Exp Med. 2021 Jun;30(6):599-605. doi: 10.17219/acem/133485.
Leukemic stem cells (LSCs) play an important role in the pathogenesis of leukemia. This research attempted to clarify effects of the telomere system on ginsenoside Rg1-induced senescence of LSCs.
This research attempted to clarify effects of the telomere system on ginsenoside Rg1-induced senescence of LSCs.
CD34+CD38- LSCs were isolated, sorted, and divided into a control group and a Rg1 group (treated with 40 μmol/L Rg1). Cell Counting Kit-8 (CCK-8) was used to evaluate cell proliferation, and flow cytometry was used to assess the cell cycle of CD34+CD38- LSCs. The senescence-associated β-galactosidase (SA-β-Gal) staining and CFU-Mix assay were conducted to measure senescence of CD34+CD38- LSCs. The mRNA transcription and protein expression of p16INK4a and human telomerase reverse transcriptase (hTERT) were determined using a real-time polymerase chain reaction (RT-PCR) and western blot assay, respectively.
The Rg1 treatment significantly attenuated proliferative activity and decreased the proliferative index (PI) of CD34+CD38- LSCs compared to those of the control group (p < 0.05). It remarkably increased positive SA-β-Gal staining rate, and suppressed formation of the CFU-Mix of CD34+CD38- LSCs compared with those of the control group (p < 0.05). The Rg1 treatment markedly boosted telomere effector, p16INK4a, in CD34+CD38- LSCs compared with that of control group (p < 0.05). Such treatment obviously reduced telomere regulator, hTERT, in CD34+CD38- LSCs compared with the control group (p < 0.05).
Ginsenoside Rg1-induced senescence of CD34+CD38- LSCs through upregulating p16INK4a and downregulating hTERT expression, both of which are associated with telomere systems. The present study would be beneficial for the treatment of leukemia by providing a promising strategy to induce senescence of CD34+CD38- LSCs.
白血病干细胞（LSCs）在白血病的发病机制中起着重要作用。本研究试图阐明端粒系统对人参皂苷 Rg1 诱导 LSCs 衰老的影响。
本研究试图阐明端粒系统对人参皂苷 Rg1 诱导 LSCs 衰老的影响。
分离、分选 CD34+CD38-LSCs，分为对照组和 Rg1 组（用 40μmol/L Rg1 处理）。用细胞计数试剂盒-8（CCK-8）评估细胞增殖，用流式细胞术评估 CD34+CD38-LSCs 的细胞周期。用衰老相关β-半乳糖苷酶（SA-β-Gal）染色和 CFU-Mix 测定评估 CD34+CD38-LSCs 的衰老情况。采用实时聚合酶链反应（RT-PCR）和 Western blot 检测 p16INK4a 和人端粒酶逆转录酶（hTERT）的 mRNA 转录和蛋白表达。
与对照组相比，Rg1 处理显著减弱了 CD34+CD38-LSCs 的增殖活性和增殖指数（PI）（p<0.05）。它显著增加了阳性 SA-β-Gal 染色率，并抑制了 CD34+CD38-LSCs 的 CFU-Mix 形成，与对照组相比（p<0.05）。与对照组相比，Rg1 处理显著增加了 CD34+CD38-LSCs 中的端粒效应物 p16INK4a（p<0.05）。这种处理明显减少了 CD34+CD38-LSCs 中的端粒调节剂 hTERT，与对照组相比（p<0.05）。
人参皂苷 Rg1 通过上调 p16INK4a 和下调 hTERT 表达诱导 CD34+CD38-LSCs 衰老，这两者都与端粒系统有关。本研究为通过诱导 CD34+CD38-LSCs 衰老来治疗白血病提供了一种有前途的策略，具有重要意义。
