Cen Hong-Xia, Cai Si-Ming, Jiang Hong-Yu, Liao Zhao-Mei, Han Dong-Guang
Department of Pediatrics, The Second Affiliated Hospital of Hainan Medical University, Haikou 570311, Hainan Province, China.
Department of Pediatrics, The Second Affiliated Hospital of Hainan Medical University, Haikou 570311, Hainan Province, China. E-mail:
Zhongguo Shi Yan Xue Ye Xue Za Zhi. 2023 Apr;31(2):344-351. doi: 10.19746/j.cnki.issn.1009-2137.2023.02.006.
To explore the effect of abnormal miRNA expression on the proliferation of pediatric acute lymphoblastic leukemia (ALL) cells and its related mechanism.
15 children with ALL and 15 healthy subjects were collected from the Second Affiliated Hospital of Hainan Medical University from July 2018 to March 2021. MiRNA sequencing was performed on their bone marrow cells, and validated using qRT-PCR. MiR-1294 and miR-1294-inhibitory molecule (miR-1294-inhibitor) were transfected into Nalm-6 cells, and the proliferation of Nalm-6 cells was detected by CCK-8 and colony formation assays. Western blot and ELISA were used to detect apoptosis of Nalm-6 cells. Biological prediction of miR-1294 was performed to find the target gene, which was verified by luciferase reporter assay. Si- was transfected into Nalm-6 cells, Western blot was used to detect the expression of Wnt signaling pathway-related proteins and to verify the effect of si- on the proliferation and apoptosis of Nalm-6 cells.
Compared with healthy subjects, 22 miRNAs were significantly upregulated in bone marrow cells of ALL patients, of which miR-1294 was the most significantly upregulated. In addition, the expression level of gene was significantly reduced in bone marrow cells of ALL patients. Compared with the NC group, the miR-1294 group showed increased protein expression levels of Wnt3a and β-catenin, faster cell proliferation, and more colony-forming units, while caspase-3 protein expression level and cell apoptosis were reduced. Compared with the NC group, the miR-1294-inhibitor group showed reduced protein expression levels of Wnt3a and β-catenin, slower cell proliferation, and fewer colony-forming units, while caspase-3 protein expression level was increased and apoptosis rate was elevated. miR-1294 had a complementary base-pair with the 3'UTR region of , and miR-1294 directly targeted . The expression of miR-1294 was negatively correlated with in ALL cells. Compared with the si-NC group, the si- group showed increased protein expression levels of Wnt3a and β-catenin, accelerated cell proliferation, and decreased caspase-3 protein expression level and cell apoptosis rate.
MiR-1294 can target and inhibit expression, thus activating the Wnt/β-Catenin signaling pathway to promote the proliferation of ALL cells, inhibit cell apoptosis, and ultimately affect the disease progression.
探讨异常miRNA表达对小儿急性淋巴细胞白血病（ALL）细胞增殖的影响及其相关机制。
收集2018年7月至2021年3月在海南医学院第二附属医院就诊的15例ALL患儿及15例健康对照者。对其骨髓细胞进行miRNA测序，并采用qRT-PCR进行验证。将miR-1294及miR-1294抑制分子（miR-1294-inhibitor）转染至Nalm-6细胞，采用CCK-8法和集落形成试验检测Nalm-6细胞的增殖情况。采用Western blot和ELISA法检测Nalm-6细胞的凋亡情况。对miR-1294进行生物学预测以寻找靶基因，并通过荧光素酶报告基因试验进行验证。将si-转染至Nalm-6细胞，采用Western blot检测Wnt信号通路相关蛋白的表达，验证si-对Nalm-6细胞增殖及凋亡的影响。
与健康对照者相比，ALL患儿骨髓细胞中有22种miRNA显著上调，其中miR-1294上调最为明显。此外，ALL患儿骨髓细胞中 基因的表达水平显著降低。与NC组相比，miR-1294组Wnt3a和β-连环蛋白的蛋白表达水平升高，细胞增殖加快，集落形成单位增多，而caspase-3蛋白表达水平及细胞凋亡减少。与NC组相比，miR-1294-inhibitor组Wnt3a和β-连环蛋白的蛋白表达水平降低，细胞增殖减慢，集落形成单位减少，而caspase-3蛋白表达水平升高，凋亡率升高。miR-1294与 的3'UTR区域存在互补碱基对，miR-129直接靶向 。ALL细胞中miR-1294的表达与 呈负相关。与si-NC组相比，si-组Wnt3a和β-连环蛋白的蛋白表达水平升高，细胞增殖加速，caspase-3蛋白表达水平及细胞凋亡率降低。
MiR-1294可靶向抑制 表达，从而激活Wnt/β-连环蛋白信号通路，促进ALL细胞增殖，抑制细胞凋亡，最终影响疾病进展。
