Yang Yin, Mei Chen, Xian Hong, Zhang Xue, Li Jun, Liang Zhi-Xuan, Zhi Yan, Ma Yue, Wang Hong-Jun
Institute of Animal Husbandry and Veterinary Medicine, Beijing Municipal Academy of Agriculture and Forestry, Beijing, China.
School of Veterinary Medicine, Southwest University, Rongchang Chongqing, China.
Vet Comp Oncol. 2023 Jun;21(2):315-326. doi: 10.1111/vco.12889. Epub 2023 Mar 8.
Toosendanin (TSN) is an active compound from the fruit of Melia toosendan Sieb et Zucc. TSN has been shown to have broad-spectrum anti-tumour activities in human cancers. However, there are still many gaps in the knowledge of TSN on canine mammary tumours (CMT). CMT-U27 cells were used to select the optimal acting time and best concentration of TSN to initiate apoptosis. Cell proliferation, cell colony formation, cell migration and cell invasion were analysed. The expression of apoptosis-related genes and proteins were also detected to explore the mechanism of action of TSN. A murine tumour model was established to detect the effect of TSN treatments. The results showed that TSN decreased cell viability of migration and invasion, altered CMT-U27 cell morphology, and inhibited DNA synthesis. TSN-induced cell apoptosis by upregulating BAX, cleaved caspase-3, cleaved caspase-9, p53 and cytochrome C (cytosolic) protein expression, and downregulating Bcl-2 and cytochrome C (mitochondrial) expression. In addition, TSN increased the mRNA transcription levels of cytochrome C, p53 and BAX, and decreased the mRNA expression of Bcl-2. Furthermore, TSN inhibited the growth of CMT xenografts by regulating the expression of genes and proteins activated by the mitochondrial apoptotic pathway. In conclusion, TSN effectively inhibited cell proliferation, migration and invasion activity, as well as induced CMT-U27 cell apoptosis. The study provides a molecular basis for the development of clinical drugs and other therapeutic options.
川楝素（TSN）是从川楝（Melia toosendan Sieb et Zucc.）果实中提取的一种活性化合物。TSN已被证明在人类癌症中具有广谱抗肿瘤活性。然而，在川楝素对犬乳腺肿瘤（CMT）的研究方面仍存在许多空白。本研究使用CMT-U27细胞来筛选川楝素诱导细胞凋亡的最佳作用时间和最佳浓度。分析细胞增殖、细胞集落形成、细胞迁移和细胞侵袭情况。检测凋亡相关基因和蛋白的表达，以探索川楝素的作用机制。建立小鼠肿瘤模型以检测川楝素治疗的效果。结果表明，川楝素降低了细胞的迁移和侵袭活力，改变了CMT-U27细胞形态，并抑制了DNA合成。川楝素通过上调BAX、裂解的caspase-3、裂解的caspase-9、p53和细胞色素C（胞质）蛋白表达，下调Bcl-2和细胞色素C（线粒体）表达来诱导细胞凋亡。此外，川楝素增加了细胞色素C、p53和BAX的mRNA转录水平，降低了Bcl-2的mRNA表达。此外，川楝素通过调节线粒体凋亡途径激活的基因和蛋白表达来抑制CMT异种移植瘤的生长。总之，川楝素有效抑制细胞增殖、迁移和侵袭活性，并诱导CMT-U27细胞凋亡。该研究为临床药物开发和其他治疗选择提供了分子基础。
