Niu Peiguang, Li Danyun, Chen Huajiao, Zhu Yanting, Zhou Jintuo, Zhang Jinhua, Liu Ying
Department of Pharmacy, Fujian Maternity and Child Health Hospital, College of Clinical Medicine for Obstetrics and Gynecology and Pediatrics, Fujian Medical University, Fuzhou, Fujian, China.
Fujian Key Laboratory of Women and Children's Critical Diseases Research [Fujian Maternity and Child Health Hospital (Fujian Women and Children's Hospital)], Fujian Maternity and Child Health Hospital, Fuzhou, Fujian, China.
PLoS One. 2025 May 2;20(5):e0322733. doi: 10.1371/journal.pone.0322733. eCollection 2025.
Metabolic reprogramming is a hallmark of cancer and de novo lipogenesis (DNL) accelerates the progression of ovarian cancer. In this study, we investigated the effects of cardamonin, a natural compound potential to suppress various malignancies, on the lipid anabolism in ovarian cancer. Cell proliferation was assessed using CCK-8 and clone formation assay. Cell apoptosis was detected by flow cytometry with Annexin V-FITC/PI staining and mitochondrial membrane potential (MMP) was measured with JC-10 probe. Free fatty acids (FFA) was measured by fluorescence using acyl-CoA oxidation and carnitine palmitoyl transferase-1 (CPT-1) activity was analyzed by spectrophotometric assay using palmitoyl-CoA and DTNB (5,5'-dithio-bis-(2-nitrobenzoic acid)) reaction. mRNA expression was measured by Quantitative Real-Time PCR. Protein expression was analyzed through western blotting and immunofluorescence. Raptor was knocked down by shRNA and Raptor was overexpressed by lentiviral transfection. The antitumor effect of cardamonin was evaluated using a xenotransplantation tumor bearing mouse model. Cardamonin suppressed the cell proliferation, induced cell apoptosis and triggered mitochondrial damage in ovarian cancer cells. Cardamonin inhibited the protein expression of sterol regulatory element binding protein 1 (SREBP1) and its downstream lipogenic enzymes and decreased FFA content and CPT-1 activity. Additionally, cardamonin inhibited the activation of mechanistic target of rapamycin complex 1 (mTORC1) and expression of regulatory-associated protein of mTOR (Raptor). Raptor knockdown abolished the inhibitory effect of cardamonin on mTORC1 and SREBP1. Furthermore, cardamonin inhibited mTORC1 activation and lipogenic proteins expression induced by Raptor overexpression. Cardamonin reduced the tumor growth and fatty acid synthase of the tumors, as evidenced by decreased expression of Ki-67 and FASN. It suggests that cardamonin suppresses mTORC1/SREBP1 through reducing the protein level of Raptor and inhibits DNL of ovarian cancer.
代谢重编程是癌症的一个标志，而从头脂肪生成（DNL）会加速卵巢癌的进展。在本研究中，我们调查了小豆蔻明（一种具有抑制多种恶性肿瘤潜力的天然化合物）对卵巢癌脂质合成代谢的影响。使用CCK-8和克隆形成试验评估细胞增殖。通过Annexin V-FITC/PI染色的流式细胞术检测细胞凋亡，并用JC-10探针测量线粒体膜电位（MMP）。使用酰基辅酶A氧化通过荧光法测量游离脂肪酸（FFA），并使用棕榈酰辅酶A和DTNB（5,5'-二硫代双-（2-硝基苯甲酸））反应通过分光光度法分析肉碱棕榈酰转移酶-1（CPT-1）活性。通过定量实时PCR测量mRNA表达。通过蛋白质免疫印迹和免疫荧光分析蛋白质表达。通过shRNA敲低Raptor，并通过慢病毒转染使Raptor过表达。使用荷瘤异种移植小鼠模型评估小豆蔻明的抗肿瘤作用。小豆蔻明抑制卵巢癌细胞的增殖，诱导细胞凋亡并引发线粒体损伤。小豆蔻明抑制固醇调节元件结合蛋白1（SREBP1）及其下游生脂酶的蛋白质表达，并降低FFA含量和CPT-1活性。此外，小豆蔻明抑制雷帕霉素复合物1（mTORC1）的机制靶点的激活以及mTOR调节相关蛋白（Raptor）的表达。敲低Raptor消除了小豆蔻明对mTORC1和SREBP1的抑制作用。此外，小豆蔻明抑制由Raptor过表达诱导的mTORC1激活和生脂蛋白表达。小豆蔻明减少了肿瘤生长和肿瘤的脂肪酸合酶，这通过Ki-67和FASN表达的降低得以证明。这表明小豆蔻明通过降低Raptor的蛋白质水平抑制mTORC1/SREBP1，并抑制卵巢癌的DNL。
