Yang Shuo, Liu Shasha, Dai Zixun
Department of Orthopedics & Soft Tissue, The Affiliated Cancer Hospital of Xiangya School of Medicine, Central South University/Hunan Cancer Hospital, No. 283, Tongzipo Road, Yuelu District, Changsha, 410013, Hunan Province, People's Republic of China.
Department of Comprehensive Bone and Joint Rehabilitation, Hunan Provincial Rehabilitation Hospital, Changsha, 410007, Hunan Province, People's Republic of China.
Cancer Chemother Pharmacol. 2024 Dec 30;95(1):15. doi: 10.1007/s00280-024-04733-w.
Polygala tenuifolia and its active components have been revealed to possess anti-tumor activities. However, the role of Tenuigenin (TEN), a bioactive ingredient from Polygala tenuifolia, in tumors such as osteosarcoma (OS) remains unclear. The present research intended to explore the efficacy and underlying mechanism of TEN on OS.
OS cells were administrated with different concentrations of TEN. Cell viability, proliferation, invasion, and migration were assessed with CCK-8 assay, colony formation assay, transwell assay, and wound healing assay, respectively. Protein and mRNA levels were determined with western blot and qRT-PCR, while protein phosphatase 2A (PP2A) activity was tested with PP2A phosphatase assay kit. The interaction between PP2A and cancerous inhibitor of protein phosphatase 2A (CIP2A) or nuclear factor kappaB (NF-κB) signaling was detected using co-immunoprecipitation. p-p65 expression in the nucleus was determined with immunofluorescence. The efficacy of TEN in vivo was also explored in a xenograft tumor model. Immunohistochemistry was performed to detect CIP2A and Ki67 in mice.
TEN treatment or CIP2A depletion repressed cell viability, proliferation, invasion, and migration in OS cells. Additionally, TEN reduced CIP2A, increased PP2A activity, and inactivated NF-κB signaling. PP2A directly interacted with CIP2A or NF-κB signaling, and PP2A inhibition reversed CIP2A knockdown-induced repression of NF-κB signaling. CIP2A overexpression overturned the efficacy of TEN, which was reversed by NF-κB inhibition. TEN decreased CIP2A, elevated PP2A activity, inactivated NF-κB signaling, and inhibited tumor growth in vivo, which was antagonized by CIP2A overexpression.
TEN suppressed OS growth via CIP2A/PP2A/NF-κB axis, indicating that it would be a novel drug for treating OS.
已揭示远志及其活性成分具有抗肿瘤活性。然而，远志的生物活性成分细叶远志皂苷（TEN）在骨肉瘤（OS）等肿瘤中的作用仍不清楚。本研究旨在探讨TEN对OS的疗效及潜在机制。
用不同浓度的TEN处理OS细胞。分别用CCK-8法、集落形成试验、Transwell试验和伤口愈合试验评估细胞活力、增殖、侵袭和迁移能力。用蛋白质印迹法和qRT-PCR测定蛋白质和mRNA水平，同时用PP2A磷酸酶检测试剂盒检测蛋白磷酸酶2A（PP2A）活性。用免疫共沉淀法检测PP2A与蛋白磷酸酶2A癌性抑制剂（CIP2A）或核因子κB（NF-κB）信号之间的相互作用。用免疫荧光法测定细胞核中p-p65的表达。还在异种移植瘤模型中探讨了TEN在体内的疗效。进行免疫组织化学检测小鼠体内的CIP2A和Ki67。
TEN处理或CIP2A缺失可抑制OS细胞的活力、增殖、侵袭和迁移。此外，TEN降低CIP2A水平，增加PP2A活性，并使NF-κB信号失活。PP2A直接与CIP2A或NF-κB信号相互作用，PP2A抑制可逆转CIP2A敲低诱导的NF-κB信号抑制。CIP2A过表达可推翻TEN的疗效，而NF-κB抑制可逆转这一疗效。TEN降低CIP2A水平，提高PP2A活性，使NF-κB信号失活，并在体内抑制肿瘤生长，而CIP2A过表达可拮抗这一作用。
TEN通过CIP2A/PP2A/NF-κB轴抑制OS生长，表明它可能是一种治疗OS的新型药物。
