Wu Rong, Xi Zhichao, Liu Mengfan, Ren Hangui, Dai Rongchen, Jiang Xue, Nik Nabil Wan Najbah, Wang Yalin, Feng Jiling, Chai Qiong, Dong Qihan, Xu Hongxi
School of Pharmacy, Shanghai University of Traditional Chinese Medicine, Shanghai, 201203, China.
Engineering Research Center of Shanghai Colleges for TCM New Drug Discovery, Shanghai, 201203, China.
Chin Med. 2023 May 28;18(1):61. doi: 10.1186/s13020-023-00774-0.
Pancreatic cancer (PAC), a malignancy that is fatal and commonly diagnosed at a late stage. Despite considerable advancements in cancer treatment, the survival rate of PAC remains largely consistent for the past 60 years. The traditional Chinese medicine formula Pulsatilla Decoction (PD) has been clinically used to treat inflammatory diseases for millennia and recently as a supplementary anti-cancer treatment in China. However, the bioactive ingredients and mechanisms underlying its anti-cancer effect remains unclear.
The composition and quality control of PD were verified through analysis by high performance liquid chromatography. Cell viability was determined using Cell Counting Kit-8 assay. The cell cycle distribution was analyzed through PI staining and flow cytometry analysis, while apoptotic cells were measured by double staining with Annexin V-FITC and PI. We used immunoblotting to examine protein expressions. The in vivo effects of β-peltatin and podophyllotoxin were evaluated on a subcutaneously-xenografted BxPC-3 cell nude mice model.
The current study demonstrated that PD markedly inhibited PAC cell proliferation and triggered their apoptosis. Four herbal PD formula was then disassembled into 15 combinations of herbal ingredients and a cytotoxicity assay showed that the Pulsatillae chinensis exerted the predominant anti-PAC effect. Further investigation indicated that β-peltatin was potently cytotoxic with IC of ~ 2 nM. β-peltatin initially arrested PAC cells at G2/M phase, followed by apoptosis induction. Animal study confirmed that β-peltatin significantly suppressed the growth of subcutaneously-implanted BxPC-3 cell xenografts. Importantly, compared to podophyllotoxin that is the parental isomer of β-peltatin but clinically obsoleted due to its severe toxicity, β-peltatin exhibited stronger anti-PAC effect and lower toxicity in mice.
Our results demonstrate that Pulsatillae chinensis and particularly its bioactive ingredient β-peltatin suppress PAC by triggering cell cycle arrest at G2/M phase and apoptosis.
胰腺癌（PAC）是一种致命的恶性肿瘤，通常在晚期才被诊断出来。尽管癌症治疗取得了显著进展，但在过去60年里，PAC的生存率基本保持不变。中药白头翁汤（PD）在临床上用于治疗炎症性疾病已有数千年历史，最近在中国也作为辅助抗癌治疗药物使用。然而，其抗癌作用的生物活性成分和机制尚不清楚。
通过高效液相色谱分析验证PD的成分和质量控制。使用细胞计数试剂盒-8法测定细胞活力。通过PI染色和流式细胞术分析细胞周期分布，同时用Annexin V-FITC和PI双染法检测凋亡细胞。我们用免疫印迹法检测蛋白质表达。在皮下异种移植BxPC-3细胞裸鼠模型上评估β-白屈菜红碱和鬼臼毒素的体内作用。
本研究表明，PD显著抑制PAC细胞增殖并引发其凋亡。然后将四种草药PD配方分解为15种草药成分组合，细胞毒性试验表明白头翁发挥了主要的抗PAC作用。进一步研究表明，β-白屈菜红碱具有很强的细胞毒性，IC约为2 nM。β-白屈菜红碱最初使PAC细胞停滞在G2/M期，随后诱导凋亡。动物研究证实，β-白屈菜红碱显著抑制皮下植入的BxPC-3细胞异种移植物的生长。重要的是，与β-白屈菜红碱的母体异构体鬼臼毒素相比，鬼臼毒素因毒性严重而在临床上已被淘汰，β-白屈菜红碱在小鼠中表现出更强的抗PAC作用和更低的毒性。
我们的结果表明，白头翁尤其是其生物活性成分β-白屈菜红碱通过引发细胞周期停滞在G2/M期和凋亡来抑制PAC。
