Chu Yanchen, Fang Yuan, Chi Jingwei, Li Jing, Zhang Dongyang, Zou Yunwen, Wang Zhijie
Department of Spinal Surgery, The Affiliated Hospital of Qingdao University, Qingdao, Shandong, China.
Department of Joint Surgery, The Affiliated Hospital of Qingdao University, Qingdao, Shandong, China.
Braz J Med Biol Res. 2018 Nov 14;51(12):e7665. doi: 10.1590/1414-431X20187665.
Osteosarcoma (OS) has a high incidence, malignity, and frequency of recurrence and metastasis. In this study, we aimed to explore the potential anti-cancer effects of Astragalus polysaccharides (APS) on human OS MG63 cells as well as underlying mechanisms. Viability of MG63 cells was assessed by CCK-8 assay to determine the adequate concentration of APS. Then, effects of APS on MG63 cell proliferation, cell cycle distribution, apoptosis, and migration and invasion were analyzed by BrdU incorporation, PI staining, flow cytometry, and transwell assays, respectively. The expression levels of proteins involved in these physiological processes were assessed by western blot analysis. Afterwards, miR-133a level in APS-treated cells was determined by qRT-PCR, and whether APS affected MG63 cells through regulation of miR-133a was determined. Finally, the activation of c-Jun N-terminal protein kinase (JNK) pathway was detected. We found that APS treatment suppressed the viability, proliferation, migration, and invasion of MG63 cells, as well as induced cell apoptosis. Moreover, APS enhanced the expression of miR-133a in MG63 cells. Knockdown of miR-133a reversed the APS treatment-induced MG63 cell proliferation, migration and invasion inhibition, as well as cell apoptosis. Furthermore, APS inactivated JNK pathway in MG63 cells. Knockdown of miR-133a reversed the APS treatment-induced inactivation of JNK pathway in MG63 cells. To conclude, APS repressed proliferation, migration, and invasion while induced apoptosis of OS MG63 cells by up-regulating miR-133a and then inactivating JNK pathway.
骨肉瘤（OS）具有高发病率、高恶性程度以及高复发和转移率。在本研究中，我们旨在探讨黄芪多糖（APS）对人骨肉瘤MG63细胞的潜在抗癌作用及其潜在机制。通过CCK-8法评估MG63细胞的活力以确定APS的合适浓度。然后，分别通过BrdU掺入法、PI染色法、流式细胞术和Transwell实验分析APS对MG63细胞增殖、细胞周期分布、凋亡以及迁移和侵袭的影响。通过蛋白质印迹分析评估参与这些生理过程的蛋白质的表达水平。之后，通过qRT-PCR测定APS处理细胞中miR-133a的水平，并确定APS是否通过调节miR-133a影响MG63细胞。最后，检测c-Jun氨基末端蛋白激酶（JNK）通路的激活情况。我们发现，APS处理可抑制MG63细胞的活力、增殖、迁移和侵袭，并诱导细胞凋亡。此外，APS可增强MG63细胞中miR-133a的表达。敲低miR-133a可逆转APS处理诱导的MG63细胞增殖、迁移和侵袭抑制以及细胞凋亡。此外，APS可使MG63细胞中的JNK通路失活。敲低miR-133a可逆转APS处理诱导的MG63细胞中JNK通路的失活。总之，APS通过上调miR-133a进而使JNK通路失活，从而抑制骨肉瘤MG63细胞的增殖、迁移和侵袭，并诱导其凋亡。
