Gong Jiangjiang, Zheng Shunli, Zhang Lei, Wang Yi, Meng Jiali
Department of Intensive Care, First Affiliated Hospital of Jinan University, Guangzhou, Guangdong, China (mainland).
First Class Ward, First Affiliated Hospital of Jinan University, Guangzhou, Guangdong, China (mainland).
Med Sci Monit. 2017 Jan 10;23:144-150. doi: 10.12659/msm.899214.
BACKGROUND Chronic myelogenous leukemia (CML) has unsatisfactory treatment efficacy at present. As the major component of red orpiment, tetra-arsenic tetra-sulfide (As4S4) has been recently used in treating leukemia, but with unclear mechanism targeting CML. MicroRNA (miR) is a group of endogenous non-coding RNAs regulating pathogenesis. MiR181 has been shown to exert important roles in tumor progression. The relationship between miR181 and As4S4 in inducing K562 cell apoptosis, however, is still unclear. MATERIAL AND METHODS CML cell line K562 was cultured in vitro in a control group and in groups receiving various dosages (20 μM and 40 μM) of As4S4. MTT assay was employed to detect the effect on K562 cell survival. MiR181 expression was quantified by real-time PCR. MTT assay and assay kit were used to determine K562 cell survival and caspase 3 expression. Cell apoptosis was assessed by flow cytometry. Bcl-2 expression was determined by real-time PCR and Western blotting. RESULTS As4S4 significantly suppressed proliferation of K562 cells (p<0.05) and decreased miR181 expression, and increased caspase3 activity compared to the control group. It can induce K562 cell apoptosis via remarkably down-regulating mRNA and protein expressions of Bcl-2 (p<0.05). CONCLUSIONS As4S4 can facilitate K562 cell apoptosis via down-regulating miR181, inhibiting Bcl02 expression, and enhancing apoptotic protein caspase3 activity.
背景 目前慢性髓性白血病(CML)的治疗效果不尽人意。作为雄黄的主要成分,四硫化四砷(As4S4)最近已用于治疗白血病,但其针对CML的作用机制尚不清楚。微小RNA(miR)是一组调节发病机制的内源性非编码RNA。已证明miR181在肿瘤进展中发挥重要作用。然而,miR181与As4S4在诱导K562细胞凋亡中的关系仍不清楚。
材料与方法 CML细胞系K562在体外培养,分为对照组和接受不同剂量(20 μM和40 μM)As4S4的组。采用MTT法检测对K562细胞存活的影响。通过实时PCR定量miR181表达。使用MTT法和试剂盒测定K562细胞存活和半胱天冬酶3表达。通过流式细胞术评估细胞凋亡。通过实时PCR和蛋白质印迹法测定Bcl-2表达。
结果 与对照组相比,As4S4显著抑制K
562细胞的增殖(p<0.05),降低miR181表达,并增加半胱天冬酶3活性。它可通过显著下调Bcl-2的mRNA和蛋白质表达(p<0.05)诱导K562细胞凋亡。
结论 As4S4可通过下调miR181、抑制Bcl-2表达和增强凋亡蛋白半胱天冬酶3活性促进K562细胞凋亡。