Liang Haihai, Li Xuelian, Wang Lu, Yu Shaonan, Xu Zhidan, Gu Yunyan, Pan Zhenwei, Li Tianyu, Hu Meiyu, Cui Hairong, Liu Xue, Zhang Ying, Xu Chaoqian, Guo Rui, Lu Yanjie, Yang Baofeng, Shan Hongli
Department of Pharmacology (the State-Province Key Laboratories of Biomedicine-Pharmaceutics of China, Harbin, P. R. China.
Cell Physiol Biochem. 2013;32(6):1818-29. doi: 10.1159/000356615. Epub 2013 Dec 16.
Arsenic trioxide (As2O3), an ancient drug used in traditional Chinese medicine, has substantial anticancer activities, especially in the treatment of patients suffering from acute promyelocytic leukemia (APL); however the underlying mechanisms are not well understood.
MTT assay was used to detect the cell viability. Flow Cytometry analysis and caspase-3 activity assay were used to measure apoptosis of APL cells. Caspase-3 and Bax levels were analyzed by western blot and let-7d and miR-766 levels were determined by real-time RT-PCR.
As2O3 significantly inhibited cell viability and induced apoptosis in APL cells. Several microRNAs, including let-7d and miR-766, were dysregulated in APL cells treated with As2O3. The expression of caspase-3 and Bax, which are targets of let-7d and miR-766, respectively, were up-regulated in As2O3 treated cells. Transfection of let-7d and miR-766 into NB4 cells decreased the expression of caspase-3 and Bax, respectively. Correspondingly, transfection of these microRNAs increased NB4 cell viability. As2O3 induced degradation of promyelocytic leukemia (PML), and then induced the down-regulation of both let-7d and miR-766 in NB4 cells.
We construct a dysregulated microRNA network involved in As2O3-induced apoptosis in APL. Targeting this network may be a new strategy for the prevention of side effects associated with APL treatment with As2O3.
三氧化二砷（As2O3）是一种用于传统中药的古老药物，具有显著的抗癌活性，尤其是在治疗急性早幼粒细胞白血病（APL）患者方面；然而其潜在机制尚不清楚。
采用MTT法检测细胞活力。采用流式细胞术分析和半胱天冬酶 - 3活性测定法检测APL细胞凋亡。通过蛋白质免疫印迹法分析半胱天冬酶 - 3和Bax水平，通过实时逆转录聚合酶链反应测定let - 7d和miR - 766水平。
As2O3显著抑制APL细胞的活力并诱导其凋亡。在用As2O3处理的APL细胞中，包括let - 7d和miR - 766在内的几种微小RNA表达失调。分别作为let - 7d和miR - 766靶标的半胱天冬酶 - 3和Bax的表达在As2O3处理的细胞中上调。将let - 7d和miR - 766转染到NB4细胞中分别降低了半胱天冬酶 - 3和Bax的表达。相应地，转染这些微小RNA增加了NB4细胞的活力。As2O3诱导早幼粒细胞白血病（PML）降解，然后诱导NB4细胞中let - 7d和miR - 766均下调。
我们构建了一个与As2O3诱导的APL细胞凋亡相关的失调微小RNA网络。针对该网络可能是预防As2O3治疗APL相关副作用的新策略。
