Liu Jun, Huang Lei, Su Pengxiao, Song Tao, Zhang Wentao, Fan Jinzhu, Liu Yang
Department of Hand and Foot Surgery and Reparative and Reconstructive Surgery, Orthopedic Hospital of The Second Hospital of Jilin University, Changchun, Jilin 130000, P.R. China.
Department of Burns, Nanfang Hospital of Southern Medical University, Guangzhou, Guangdong 510515, P.R. China.
Oncol Lett. 2018 Apr;15(4):4113-4120. doi: 10.3892/ol.2018.7814. Epub 2018 Jan 17.
A number of studies have attempted to elucidate the association between mircoRNAs (miRNAs/miRs) and cancer-associated processes. The aim of the present study was to determine how miR-499a-5p intervenes in human osteosarcoma cell proliferation and differentiation. The cancerous tissues and adjacent non-cancerous tissues of 62 patients with osteosarcoma (OS) were collected. miRNA microarray analysis revealed that 29 miRNAs were upregulated while 26 were downregulated, among which miR-499a-5p expression was the most decreased. Western blot analysis and reverse transcription-quantitative polymerase chain reaction demonstrated that the mRNA and protein expression of miR-499a-5p was lower, while that of protein phosphatase 1D () was higher in OS tissues compared with expression levels in normal tissues. Furthermore, miR-499a-5p expression was markedly decreased in the metastatic tumors and in those at stage III+IV compared with the non-metastatic tumors and those at stage I, respectively. In addition, following transfection of the human OS MG-63 cell line with an miR-499a-5p mimic, the expression of miR-499a-5p was elevated while the protein and mRNA expression of was decreased. When combining these findings with the information obtained from the Targetscan predictive software, it was confirmed that was targeted by miR-499a-5p. In MG-63 cells transfected with an miR-499a-5p mimic, -associated downstream proteins phosphorylated protein kinase B (p-Akt) and phosphorylated glycogen synthase kinase 3β (p-GSK-3β) were significantly downregulated compared with the negative control (NC) group, while the expression of p-Akt and p-GSK-3β were significantly elevated in the tumor tissues compared with the adjacent non-tumor tissues. Simultaneously, the growth and proliferation activity of MG-63 cells were notably reduced when transfected with the miR-499a-5p mimic, compared with the NC group. Therefore, it may be concluded that miR-499a-5p suppresses OS cell proliferation and differentiation by targeting through modulation of Akt/GSK-3β signaling.
多项研究试图阐明微小RNA（miRNAs/miRs）与癌症相关过程之间的关联。本研究的目的是确定miR-499a-5p如何干预人骨肉瘤细胞的增殖和分化。收集了62例骨肉瘤（OS）患者的癌组织和相邻的非癌组织。miRNA微阵列分析显示，29种miRNA上调，26种下调，其中miR-499a-5p表达下降最为明显。蛋白质印迹分析和逆转录-定量聚合酶链反应表明，与正常组织中的表达水平相比，OS组织中miR-499a-5p的mRNA和蛋白质表达较低，而蛋白磷酸酶1D（）的表达较高。此外，与非转移性肿瘤和I期肿瘤相比，转移性肿瘤和III + IV期肿瘤中miR-499a-5p表达分别明显降低。此外，用miR-499a-5p模拟物转染人OS MG-63细胞系后，miR-499a-5p的表达升高，而的蛋白质和mRNA表达降低。将这些发现与从Targetscan预测软件获得的信息相结合，证实是miR-499a-5p的靶标。与阴性对照（NC）组相比，用miR-499a-5p模拟物转染的MG-63细胞中，与相关的下游蛋白磷酸化蛋白激酶B（p-Akt）和磷酸化糖原合酶激酶3β（p-GSK-3β）明显下调，而与相邻非肿瘤组织相比，肿瘤组织中p-Akt和p-GSK-3β的表达明显升高。同时，与NC组相比，用miR-499a-5p模拟物转染时，MG-63细胞的生长和增殖活性明显降低。因此，可以得出结论miR-499a-5p通过调节Akt/GSK-3β信号通路靶向抑制OS细胞的增殖和分化。
