Cheng Deliang, Li Jiageng, Zhang Lijun, Hu Leiming
Department of Hand Surgery, Honghui Hospital, Xi'an Jiaotong University, Xi'an, Shaanxi 710054, P.R. China.
Oncol Lett. 2019 Jan;17(1):1363-1371. doi: 10.3892/ol.2018.9712. Epub 2018 Nov 15.
Previous studies have revealed that miR-142-5p serves a critical role in human cancer progression. However, the biological function of miR-142-5p in osteosarcoma (OS) development remains unclear. In the present study, the role of miR-142-5p in human OS HOS cells was determined, and the underlying mechanism involved was examined. Compared with the adjacent healthy tissues, the expression level of miR-142-5p was downregulated and the expression level of group XVI phospholipase A2 (PLA2G16) protein was upregulated in human OS tissues. The aforementioned results were also indicated in human OS HOS cells when compared with human fetal osteoblastic hFOB1.19 cells. Additionally, the results demonstrated that PLA2G16 was a direct target of miR-142-5p. miR-142-5p transfection upregulated the expression level of miR-142-5p and suppressed the expression level of PLA2G16 protein in HOS cells. MTT assays indicated a time-dependent decrease by miR-142-5p transfection in the proliferation of HOS cells. 5-bromo-2'-deoxyuridine incorporation assays confirmed that miR-142-5p transfection inhibited DNA synthesis in HOS cells. In addition, miR-142-5p transfection increased the Caspase-3 (CASP3) activity and apoptotic rate. Western blot analysis indicated that miR-142-5p transfection reduced BCL2, apoptosis regulator expression and upregulated the expression of CASP3 and BCL2 associated X, apoptosis regulator in HOS cells. Furthermore, miR-142-5p transfection decreased the expression levels of phosphorylated (p)-proto-oncogene, serine/threonine kinase, p-mitogen-activated protein kinase kinase, and p-extracellular signal-regulated kinase (ERK) 1/2 proteins in HOS cells. PLA2G16 overexpression restored the expression level of p-ERK 1/2 protein, which was reduced by miR-142-5p overexpression. MTT and CASP3 activity assays indicated that restoration of PLA2G16 reversed the tumour-suppressive role of miR-142-5p transfection in HOS cells. In conclusion, the results of the present study indicated that miR-142-5p suppressed proliferation and promoted apoptosis in human OS HOS cells by targeting PLA2G16 through ERK1/2 signaling pathway.
先前的研究表明,miR-142-5p在人类癌症进展中起关键作用。然而,miR-142-5p在骨肉瘤(OS)发生发展中的生物学功能仍不清楚。在本研究中,确定了miR-142-5p在人骨肉瘤HOS细胞中的作用,并研究了其潜在机制。与相邻的健康组织相比,人骨肉瘤组织中miR-142-5p的表达水平下调,而第十六组磷脂酶A2(PLA2G16)蛋白的表达水平上调。与人类胎儿成骨细胞hFOB1.19细胞相比,上述结果在人骨肉瘤HOS细胞中也得到了证实。此外,结果表明PLA2G16是miR-142-5p的直接靶点。miR-142-5p转染上调了HOS细胞中miR-142-5p的表达水平,并抑制了PLA2G16蛋白的表达水平。MTT分析表明,miR-142-5p转染使HOS细胞的增殖呈时间依赖性下降。5-溴-2'-脱氧尿苷掺入试验证实,miR-142-5p转染抑制了HOS细胞中的DNA合成。此外,miR-142-5p转染增加了半胱天冬酶-3(CASP3)活性和凋亡率。蛋白质印迹分析表明,miR-142-5p转染降低了HOS细胞中凋亡调节因子BCL2的表达,并上调了CASP3和凋亡调节因子BCL2相关X蛋白的表达。此外,miR-142-5p转染降低了HOS细胞中磷酸化(p)-原癌基因、丝氨酸/苏氨酸激酶、p-丝裂原活化蛋白激酶激酶和p-细胞外信号调节激酶(ERK)1/2蛋白的表达水平。PLA2G16过表达恢复了被miR-142-5p过表达降低的p-ERK 1/2蛋白的表达水平。MTT和CASP3活性分析表明,PLA2G16的恢复逆转了miR-1