Xing Weiwei, Yin Yuxia, Yang Saina, Lu Guang
Department of Pediatrics, Yidu Central Hospital of Weifang, Weifang, Shandong 262500, P.R. China.
Department of Neurosurgery Ward II, Yidu Central Hospital of Weifang, Weifang, Shandong 262500, P.R. China.
Oncol Lett. 2020 Apr;19(4):2870-2874. doi: 10.3892/ol.2020.11396. Epub 2020 Feb 14.
Effect of gemcitabine (GEM) on proliferation and apoptosis of childhood acute leukemia (AL) cells and the mechanism of action were investigated. Bone marrow and peripheral blood of 18 newly diagnosed children with childhood AL admitted to Yidu Central Hospital of Weifang were selected, and the miR-125a-3p level in peripheral blood of healthy children and children with AL was detected by quantitative reverse transcription-polymerase chain reaction (qRT-PCR). Leukemia cells from the bone marrow of children with AL were primarily cultured and purified to observe the morphology. miR-125a-3p mimic was transfected into childhood AL cells. The cells were randomly divided into three groups: control group, GEM group and GEM + miR-125a-3p mimic group. 5-ethynyl-2'-deoxyuridine (EdU) staining assay was chosen to detect the proliferation of childhood AL cells in each group. Terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) staining assay was adopted to determine apoptosis of childhood AL cells. The protein level of c-myc was measured via western blotting. Compared with that in the healthy children, the level of miR-125a-3p in the peripheral blood of children with AL was remarkably decreased. Compared with those in the control group, GEM inhibited proliferation and promoted apoptosis of childhood AL cells, and impeded the protein expression of c-myc in these cells. Compared with those in the GEM group, GEM + miR-125a-3p mimic notably reduced the proliferation and enhanced apoptosis of cells, and the protein expression of c-myc in cells was overtly reduced. The level of miR-125a-3p in peripheral blood of children with AL is obviously decreased. It is suggested in this study that GEM can inhibit the proliferation and promote apoptosis of childhood AL cells, and the mechanism may be related to upregulated miR-125a-3p inhibiting the expression of c-myc.
研究了吉西他滨（GEM）对儿童急性白血病（AL）细胞增殖和凋亡的影响及其作用机制。选取潍坊益都中心医院收治的18例新诊断儿童AL的骨髓和外周血，采用定量逆转录-聚合酶链反应（qRT-PCR）检测健康儿童和AL患儿外周血中miR-125a-3p水平。对AL患儿骨髓中的白血病细胞进行原代培养和纯化以观察形态。将miR-125a-3p模拟物转染至儿童AL细胞中。细胞随机分为三组：对照组、GEM组和GEM + miR-125a-3p模拟物组。采用5-乙炔基-2'-脱氧尿苷（EdU）染色法检测各组儿童AL细胞的增殖情况。采用末端脱氧核苷酸转移酶dUTP缺口末端标记（TUNEL）染色法检测儿童AL细胞的凋亡情况。通过蛋白质印迹法检测c-myc的蛋白水平。与健康儿童相比，AL患儿外周血中miR-125a-3p水平显著降低。与对照组相比，GEM抑制儿童AL细胞增殖并促进其凋亡，并抑制这些细胞中c-myc的蛋白表达。与GEM组相比，GEM + miR-125a-3p模拟物显著降低细胞增殖并增强细胞凋亡，且细胞中c-myc的蛋白表达明显降低。AL患儿外周血中miR-125a-3p水平明显降低。本研究表明，GEM可抑制儿童AL细胞增殖并促进其凋亡，其机制可能与上调miR-125a-3p抑制c-myc表达有关。
