Our goal was to evaluate phytochemical characterization and the antitumor potential of Calotropis procera. The phytochemical constitution of the crude extract (CE) revealed the presence of flavonoids, glycosides and cardenolide. The MTT assay was used to evaluate the cytotoxicity of CE, methanolic (MF) and ethyl acetate fractions (EAF) of C. procera in canine osteosarcoma cells (OST), canine mammary tumor (CMT), and canine skin fibroblasts (non-tumor cell). Doxorubicin was also used as a positive control. Results showed that CE, MF and EAF promoted a decrease in the viability of OST and CMT cells and did not alter the fibroblasts viability. C. procera also decreased the number of cells, corroborating to the decrease in proliferation and the cell cycle arrest in the G0/G1 phase. It was also evaluated the cell morphology by light and fluorescence microscopy, being demonstrated a reduction in cytoplasmic and cell rounding characteristic of programmed cell death. Moreover, flow cytometry data demonstrated that CE treatment promoted increase of caspase-3 and p53, showing that the cell death was activated in OST cells. In addition, there was a decrease in CD31, VEGF, osteopontin and TGF-β after CE treatment, suggesting that CE exerts its antitumor effect by reducing angiogenesis and tumor progression in OST cells. Moreover, CMT cells showed a reduction in PCNA after treatment with MF and CE. Analyzing the data together, C. procera, especially CE, showed an antitumor potential in both OST and CMT cells, encouraging us to continue investigating its use in cancer therapy.

我们的目标是评估Calotropis procera 的植物化学特性和抗肿瘤潜力。粗提取物 (CE) 的植物化学成分表明存在类黄酮、糖苷和心内酯。MTT 测定用于评估C. procera的 CE、甲醇 (MF) 和乙酸乙酯组分 (EAF)在犬骨肉瘤细胞 (OST)、犬乳腺肿瘤 (CMT) 和犬皮肤成纤维细胞（非肿瘤细胞）中的细胞毒性细胞）。阿霉素也用作阳性对照。结果表明，CE、MF 和 EAF 促进了 OST 和 CMT 细胞活力的降低，而不会改变成纤维细胞的活力。C. procera还减少了细胞数量，证实了 G0/G1 期增殖和细胞周期停滞的减少。它还通过光学和荧光显微镜评估了细胞形态，证明了程序性细胞死亡的细胞质和细胞变圆特征的减少。此外，流式细胞术数据表明，CE 处理促进了 caspase-3 和 p53 的增加，表明 OST 细胞中的细胞死亡被激活。此外，CE治疗后CD31、VEGF、骨桥蛋白和TGF-β均降低，表明CE通过减少OST细胞的血管生成和肿瘤进展发挥其抗肿瘤作用。此外，CMT 细胞在用 MF 和 CE 处理后显示出 PCNA 减少。一起分析数据，C. procera，尤其是 CE，在 OST 和 CMT 细胞中均显示出抗肿瘤潜力，鼓励我们继续研究其在癌症治疗中的应用。