Licochalcone A-Induced Apoptosis Through the Activation of p38MAPK Pathway Mediated Mitochondrial Pathways of Apoptosis in Human Osteosarcoma Cells In Vitro and In Vivo
Licochalcone A-Induced Apoptosis Through the Activation of p38MAPK Pathway Mediated Mitochondrial Pathways of Apoptosis in Human Osteosarcoma Cells In Vitro and In Vivo
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DOI：
10.3390/cells8111441
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发表时间：
2019-11-01
期刊：
影响因子：
6
通讯作者：
Hsieh, Yi-Hsien
中科院分区：
文献类型：
--
作者：
Lin, Renn-Chia;Yang, Shun-Fa;Hsieh, Yi-Hsien
背景：甘草酮A(LICA)是从光果甘草根中分离得到的一种化合物，对多种肿瘤细胞具有抗肿瘤和抗侵袭活性。然而，无论是在体外还是在体内，LICA对人骨肉瘤细胞的抗肿瘤作用还没有得到证实。方法：采用四甲基偶氮唑盐比色法测定细胞存活率。Annexin V/PI染色和JC-1染色检测细胞凋亡和线粒体功能障碍。Western blotting检测到caspase或线粒体相关蛋白的表达。LICA对143B小鼠移植瘤的体内抗肿瘤作用。结果：LICA可抑制人骨肉瘤细胞增殖，诱导细胞凋亡，表现为细胞存活率下降，线粒体膜电位丧失，caspase激活。LICA可显著降低HOS和U2OS细胞中Bcl2和Mcl-1的表达，增加Caspase-3、Caspase-9、PARP和Bax的表达。此外，Z-VAD或牛磺熊去氧胆酸介导的线粒体膜电位和细胞凋亡抑制显著减少了LICA诱导的线粒体依赖的细胞凋亡。研究还确定，LICA处理诱导了p38MAPK的磷酸化，但siRNA-p38或BIRB796通过抑制线粒体依赖的凋亡途径显著逆转了细胞的活力。最后，体内研究表明，LICA显著抑制了143B移植瘤的生长。结论：这些结果表明LICA在体内外通过p38MAPK调节线粒体介导的内源性凋亡途径而具有抗骨肉瘤细胞活性。
Background: Licochalcone A (LicA) is isolated from the roots of Glycyrrhiza glabra and possesses antitumor and anti-invasive activities against several tumor cells. However, the antitumor effects of LicA on human osteosarcoma cells have yet to be demonstrated either in vitro or in vivo. Methods: Cell viability was measured by MTT assay. Apoptosis and mitochondrial dysfunction were detected with Annexin V/PI staining and JC-1 staining by flow cytometry. The expressions of caspase- or mitochondrial-related proteins were demonstrated by western blotting. Antitumor effect of LicA on 143B xenograft mice in vivo. Results: LicA could inhibit cell proliferation and induce apoptosis in human osteosarcoma cells, as evidenced by a decrease in cell viability, loss of mitochondrial membrane potentials, and activation of caspases. LicA treatment substantially reduced the expression of Bcl-2 and Mcl-1 and increased the expression of cleaved-caspase-3, cleaved-caspase-9, cleaved-PARP, and Bax in HOS and U2OS cells. Moreover, mitochondrial membrane potential and apoptosis suppression mediated by Z-VAD or tauroursodeoxycholic acid significantly reduced LicA-induced mitochondria-dependent apoptosis. The study also determined that LicA treatment induced p38MAPK phosphorylation, but siRNA-p38 or BIRB796 substantially reversed cell viability through the inhibition of mitochondria-dependent apoptosis pathways. Finally, an in vivo study revealed that LicA significantly inhibited 143B xenograft tumor growth. Conclusions: These findings demonstrate that LicA has antitumor activities against human osteosarcoma cells through p38MAPK regulation of mitochondria-mediated intrinsic apoptotic pathways in vitro and in vivo.