Li Quanxiu, Li Guangchun, Liu Changyi, Chen Na, Deng Bangyu, Xie Youke
Department of Orthopedics, The Third Affiliated Hospital of Shenzhen University, Shenzhen, Guangdong, People's Republic of China.
Department of Pathology, Guangdong Women and Children Hospital, Guangzhou, Guangdong, People's Republic of China.
Cancer Manag Res. 2020 Jun 15;12:4541-4548. doi: 10.2147/CMAR.S248851. eCollection 2020.
CDA-2 (cell differentiation agent 2), isolated from healthy human urine, exerts antitumor effects in multiple types of cancer cells. However, its role in osteosarcoma has not been studied.
The MTT assay was used to examine the cell proliferation rate. A colony formation assay was used to examine cell growth. The Transwell assay was used to examine cell migration ability. A real-time PCR assay was used to examine the expression levels of miR-124 and MAPK1. A Western blot assay was used to examine protein expression levels. MAPK1 was selected as a possible target of miR-124, and the targeting relationship was examined by a luciferase reporter assay.
We revealed that CDA-2 decreased the growth, migration and invasion ability of the osteosarcoma cell line Saos-2. Further study revealed that CDA-2 elevated the expression level of miR-124. MAPK1 was identified as a downstream target of miR-124. Knockdown of miR-124 or overexpression of MAPK1 counteracted CDA-2's effects on cell growth and invasion.
Our data revealed that the miR-124/MAPK1 axis mediated CDA-2's function in Saos-2 cells. CDA-2 can be used as a new treatment strategy for osteosarcoma.
从健康人尿液中分离出的细胞分化因子2(CDA-2)在多种癌细胞中发挥抗肿瘤作用。然而,其在骨肉瘤中的作用尚未得到研究。
采用MTT法检测细胞增殖率。采用集落形成试验检测细胞生长情况。采用Transwell试验检测细胞迁移能力。采用实时PCR试验检测miR-124和MAPK1的表达水平。采用蛋白质免疫印迹法检测蛋白质表达水平。选择MAPK1作为miR-124的可能靶点,并通过荧光素酶报告基因试验检测靶向关系。
我们发现CDA-2降低了骨肉瘤细胞系Saos-2的生长、迁移和侵袭能力。进一步研究发现,CDA-2提高了miR-124的表达水平。MAPK1被确定为miR-124的下游靶点。敲低miR-124或过表达MAPK1可抵消CDA-2对细胞生长和侵袭的影响。
我们的数据表明,miR-124/MAPK1轴介导了CDA-2在Saos-2细胞中的功能。CDA-2可作为骨肉瘤的一种新的治疗策略。