He Gang, Feng Yanjiao, Chen Tangcong, Zhang Yiyuan, Liang Li, Yan Jun, Song Yanxia, Chen Fengzheng, Liu Wei
Key Laboratory of Medicinal and Edible Plants Resources Development of Sichuan Education Department, Sichuan Industrial Institute of Antibiotics, School of Pharmacy, Chengdu University, Chengdu, China.
Antibiotics Research and Re-evaluation Key Laboratory of Sichuan Province, Sichuan Industrial Institute of Antibiotics, School of Pharmacy, Chengdu University, Chengdu, China.
Bioimpacts. 2025 Mar 2;15:30688. doi: 10.34172/bi.30688. eCollection 2025.
Prostate cancer (PCa) often progresses to castration-resistant prostate cancer (CRPC), which is linked to higher treatment resistance and recurrence rates. This highlights the urgent need for new therapeutic options. Natural products, especially flavonoids, have shown promise in reducing drug resistance and possess both antioxidant and anticancer effects. Developing drugs that specifically target CRPC could offer significant therapeutic advantages.
Chrysosplenetin B (CspB) was extracted and purified from the herb (DC.) Benth. using traditional flavonoid extraction techniques, followed by high-performance liquid chromatography (HPLC) for purity assessment and nuclear magnetic resonance (NMR) for structural identification. The effect of CspB on the viability of PCa cells was evaluated using the Cell Counting Kit-8 assay. Subsequently, transcriptome analysis was conducted, and cell cycle progression was assessed through flow cytometry in conjunction with propidium iodide (PI) staining. Additionally, western blotting and quantitative real-time polymerase chain reaction (qRT-PCR) were employed to confirm the expression levels of relevant proteins and genes.
CspB was found to inhibit the proliferation of PC3, DU145, and LNCaP cells in a dose-dependent manner, with a stronger effect noted in PC3 and DU145 cells. Transcriptomic analysis revealed that CspB treatment led to cell cycle arrest, particularly in PC3 cells. Flow cytometry with PI staining confirmed that CspB caused G1 phase cell cycle arrest in PC3 cells. Moreover, CspB treatment significantly increased the expression of essential members of the Cip/Kip family, including CIP1/P21 and KIP1/P27, as well as CDKN2B (P15) and CDKN2D (P19) from the INK4 family. Additionally, CspB exposure notably raised the expression of the G1 phase-negative regulatory gene , while key cell cycle regulators like CDK6 and E2F1 were significantly downregulated at the protein level.
Our findings indicate that CspB effectively inhibits the proliferation of CRPC cells by reducing the activity of cell cycle proteins and cyclin-dependent kinase (CDK) complexes while upregulating the expression of P21 and P27 and inducing G1 phase cell cycle arrest. These results highlight the potential of CspB as a promising candidate for developing therapeutic agents aimed at targeting CRPC.
前列腺癌（PCa）常进展为去势抵抗性前列腺癌（CRPC），这与更高的治疗抵抗性和复发率相关。这凸显了对新治疗方案的迫切需求。天然产物，尤其是黄酮类化合物，在降低耐药性方面显示出前景，并且具有抗氧化和抗癌作用。开发专门针对CRPC的药物可能具有显著的治疗优势。
使用传统黄酮类提取技术从（DC.）Benth.的草药中提取并纯化金腰子苷B（CspB），随后采用高效液相色谱（HPLC）进行纯度评估，并用核磁共振（NMR）进行结构鉴定。使用细胞计数试剂盒-8检测法评估CspB对PCa细胞活力的影响。随后进行转录组分析，并通过碘化丙啶（PI）染色结合流式细胞术评估细胞周期进程。此外，采用蛋白质印迹法和定量实时聚合酶链反应（qRT-PCR）来确认相关蛋白质和基因的表达水平。
发现CspB以剂量依赖性方式抑制PC3、DU145和LNCaP细胞的增殖，在PC3和DU145细胞中作用更强。转录组分析显示，CspB处理导致细胞周期停滞，尤其是在PC3细胞中。PI染色的流式细胞术证实CspB使PC3细胞在G1期发生细胞周期停滞。此外，CspB处理显著增加了Cip/Kip家族关键成员的表达，包括CIP1/P21和KIP1/P27，以及INK4家族的CDKN2B（P15）和CDKN2D（P19）。此外，CspB处理显著提高了G1期负调控基因的表达，而关键细胞周期调节因子如CDK6和E2F1在蛋白质水平上显著下调。
我们的研究结果表明，CspB通过降低细胞周期蛋白和细胞周期蛋白依赖性激酶（CDK）复合物的活性，同时上调P21和P27的表达并诱导G1期细胞周期停滞，有效抑制CRPC细胞的增殖。这些结果突出了CspB作为开发针对CRPC的治疗药物的有前景候选物的潜力。
