Baicalin, a flavonoid derivative, exerts antitumor activity in a variety of neoplasms. However, whether baicalin exerts antitumor effects on osteosarcoma cells remains to be elucidated. In this study, treatment with baicalin reduced the proliferation and invasive potential of osteosarcoma cells and reduced the mitochondrial membrane potential, which eventually caused mitochondrial apoptosis. In addition, baicalin increased intercellular Ca²⁺ and ROS concentrations. Baicalin-induced apoptosis was confirmed by enhanced Bax, cleaved caspase-3, and cleaved PARP levels and decreased Bcl-2 levels. The increase in LC3-II and p62 suggested that baicalin induced autophagosome formation but ultimately inhibited downstream autophagy. Moreover, apoptosis induced by baicalin was attenuated by the addition of 3-MA. Furthermore, we found that baicalin inhibited the PI3K/Akt/mTOR, ERK1/2 and β-catenin signaling pathways. Chelation of free Ca²⁺ by BAPTA-AM also inhibited both apoptosis induction and ROS concentration changes. Finally, NAC pretreatment reversed baicalin treatment outcomes, including the increase in Ca²⁺ concentration, induction of apoptosis and autophagy, and inhibition of the pathways. Molecular docking results indicated that baicalin might interact with the structural domain of PI3Kγ. Thus, baicalin may be considered a potential candidate for osteosarcoma treatment.

黄芩苷是一种类黄酮衍生物，在多种肿瘤中发挥抗肿瘤活性。然而，黄芩苷是否对骨肉瘤细胞发挥抗肿瘤作用仍有待阐明。在本研究中，黄芩苷治疗降低了骨肉瘤细胞的增殖和侵袭能力，降低了线粒体膜电位，最终导致线粒体凋亡。此外，黄芩苷增加细胞间Ca ²⁺和 ROS 浓度。通过增强的 Bax、切割的 caspase-3 和切割的 PARP 水平以及降低的 Bcl-2 水平证实了黄芩苷诱导的细胞凋亡。LC3-II 和 p62 的增加表明黄芩苷诱导自噬体形成，但最终抑制下游自噬。此外，黄芩苷诱导的细胞凋亡通过添加3-MA而减弱。此外，我们发现黄芩苷抑制 PI3K/Akt/mTOR、ERK1/2 和 β-catenin 信号通路。BAPTA-AM螯合游离 Ca ²⁺也抑制细胞凋亡诱导和 ROS 浓度变化。最后，NAC 预处理逆转了黄芩苷的治疗结果，包括 Ca ^2+的增加浓度、细胞凋亡和自噬的诱导以及通路的抑制。分子对接结果表明黄芩苷可能与PI3Kγ的结构域相互作用。因此，黄芩苷可能被认为是骨肉瘤治疗的潜在候选者。