Background: The microtubule protein inhibitor C118P shows excellent anti-breast cancer effects. However, the potential targets and mechanisms of C118P in breast cancer remain unknown. Methods: Real-time cellular analysis (RTCA) was used to detect cell viability. Apoptosis and the cell cycle were detected by flow cytometry. Computer docking simulations, surface plasmon resonance (SPR) technology, and microscale thermophoresis (MST) were conducted to study the interaction between C118P and alanine-serine-cysteine transporter 2 (ASCT2). Seahorse XF technology was used to measure the basal oxygen consumption rate (OCR). The effect of C118P in the adipose microenvironment was explored using a co-culture model of adipocytes and breast cancer cells and mouse cytokine chip. Results: C118P inhibited proliferation, potentiated apoptosis, and induced G2/M cell cycle arrest in breast cancer cells. Notably, ASCT2 was validated as a C118P target through reverse docking, SPR, and MST. C118P suppressed glutamine metabolism and mediated autophagy via ASCT2. Similar results were obtained in the adipocyte–breast cancer microenvironment. Adipose-derived interleukin-6 (IL-6) promoted the proliferation of breast cancer cells by enhancing glutamine metabolism via ASCT2. C118P inhibited the upregulation of ASCT2 by inhibiting the effect of IL-6 in co-cultures. Conclusion: C118P exerts an antitumour effect against breast cancer via the glutamine transporter ASCT2.
背景：微管蛋白抑制剂C118P展现出优异的抗乳腺癌活性，但其在乳腺癌中的潜在作用靶点与机制尚未明确。方法：采用实时细胞分析技术检测细胞活力，流式细胞术检测细胞凋亡与周期分布。通过计算机反向对接模拟、表面等离子共振技术及微量热泳动实验研究C118P与丙氨酸-丝氨酸-半胱氨酸转运蛋白2的相互作用。使用Seahorse XF技术检测基础耗氧率。通过脂肪细胞与乳腺癌细胞共培养模型及小鼠细胞因子芯片探究C118P在脂肪微环境中的作用。结果：C118P可抑制乳腺癌细胞增殖、促进细胞凋亡并诱导G2/M期周期阻滞。值得注意的是，通过反向对接、表面等离子共振及微量热泳动实验证实ASCT2是C118P的直接作用靶点。C118P通过靶向ASCT2抑制谷氨酰胺代谢并介导自噬过程。在脂肪细胞-乳腺癌细胞共培养微环境中观察到相似现象：脂肪源性白细胞介素-6通过ASCT2增强谷氨酰胺代谢进而促进乳腺癌细胞增殖，而C118P能抑制IL-6对ASCT2的上调作用。结论：C118P通过靶向谷氨酰胺转运蛋白ASCT2发挥抗乳腺癌作用。
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