Xu Xiqiang, Liu Mengmeng, Wu Hua
Department of Spine Surgery, Shandong Provincial Hospital Affiliated to Shandong First Medical University Jinan, Shandong, China.
Department of Anesthesiology, Shandong Second Provincial General Hospital Jinan, Shandong, China.
Am J Transl Res. 2023 Feb 15;15(2):729-744. eCollection 2023.
To investigate the effects and mechanisms of berberine (BBR) on the migration, invasion, proliferation and apoptosis of osteosarcoma cells in vitro.
Proliferation of MG-63 and U2OS cells was measured by the CCK-8 assay. Cells migration was examined by wound-healing assay. The invasion and metastasis of cells were evaluated by transwell invasion assay. Cells apoptosis was determined by the flow cytometry. Caspase-3 activity in MG-63 and U2OS cells was measured, and Western blot was used to measure the levels of Bax, Bcl-2, MMP-2 and MMP-9 in cells. In addition, the osteosarcoma graft tumor model of mice was established. The tumorigenesis of MG-63 cells in nude mice was compared among three groups. Immunohistochemistry assay was used to measure the levels of MMP-2, MMP-9 and NM-23 in tumor tissue.
It was showed that BBR inhibited the proliferation of MG-63 and U2OS cells in vitro in time- and concentration-dependent manners. Moreover, BBR reduced the cells migration and invasion, also down-regulated the expressions of MMP-2 and MMP-9. BBR also inhibited the cells apoptosis by down-regulating the expression of Bcl-2 and up-regulating the expression of Bax. In nude mice, BBR obviously inhibited the tumorigenesis of MG-63 cells. Compared with the negative group, BBR decreased the levels of MMP-2 and MMP-9 and increased the level of NM-23. The molecular mechanism was associated with activation of the MAPK/JNK signal transduction pathway.
BBR significantly regulates the biological behaviors of osteosarcoma cells and inhibits the growth of osteosarcoma. The molecular mechanism may be associated with the modulation of MMP/NM-23 and MAPK/JNK signals. BBR may be a potential drug for the treatment of osteosarcoma.
探讨黄连素（BBR）对骨肉瘤细胞体外迁移、侵袭、增殖及凋亡的影响及其机制。
采用CCK-8法检测MG-63和U2OS细胞的增殖情况。采用划痕实验检测细胞迁移能力。采用Transwell侵袭实验评估细胞的侵袭和转移能力。采用流式细胞术检测细胞凋亡情况。检测MG-63和U2OS细胞中Caspase-3活性，并用蛋白质免疫印迹法检测细胞中Bax、Bcl-2、MMP-2和MMP-9的水平。此外，建立小鼠骨肉瘤移植瘤模型。比较三组裸鼠中MG-63细胞的成瘤情况。采用免疫组织化学法检测肿瘤组织中MMP-2、MMP-9和NM-23的水平。
结果显示，BBR能以时间和浓度依赖性方式抑制MG-63和U2OS细胞的体外增殖。此外，BBR可降低细胞的迁移和侵袭能力，同时下调MMP-2和MMP-9的表达。BBR还通过下调Bcl-2的表达和上调Bax的表达来抑制细胞凋亡。在裸鼠中，BBR明显抑制MG-63细胞的成瘤。与阴性组相比，BBR降低了MMP-2和MMP-9的水平，提高了NM-23的水平。分子机制与MAPK/JNK信号转导通路的激活有关。
BBR可显著调节骨肉瘤细胞的生物学行为，抑制骨肉瘤生长。其分子机制可能与调节MMP/NM-23和MAPK/JNK信号有关。BBR可能是一种治疗骨肉瘤的潜在药物。
