Hou Jia, Kang Ning, Liu Nan-Nan, Tan Dan, Zhang Si, Liu Jing, Xie Youhua
Key Laboratory of Medical Molecular Virology (NHC & MOE & CAMS), Department of Medical Microbiology and Parasitology, School of Basic Medical Sciences, Shanghai Institute of Infectious Diseases and Biosecurity, Shanghai Medical College, Fudan University, Shanghai, China.
Department of Biochemistry and Molecular Biology, NHC Key Laboratory of Glycoconjugates Research, School of Basic Medical Sciences, Fudan University, Shanghai, China.
Ann Transl Med. 2022 Aug;10(15):820. doi: 10.21037/atm-22-1085.
Pancreatic cancer (PC) is a highly metastatic and lethal cancer with a very low overall 5-year survival rate. There is an urgent need for identifying new therapeutic agents for this deadly disease. Cardiac glycosides (CGs) have been traditionally used for their potent cardiovascular activities and have also recently been reported to exhibit anti-tumor effects. Proscillaridin A (Pro A), a natural CG, has been shown to display anti-tumor effects on multiple cancer types.
The cytotoxic effect of Pro A on PC cells was determined using cell viability assay, colony formation assay and transwell assay . Cell apoptosis, cell cycle, reactive oxygen species (ROS) generation, intracellular Ca levels and mitochondrial membrane potential (MMP) were assayed by flow cytometry. Panc-1-xenografted mice model was used to evaluate Pro A's effect in tumor growth. Mitochondria morphology was observed by transmission electron microscopy. LC3 aggregation was assessed by GFP-LC3 fluorescence microscopy. Gene expression was assayed by western blot or real-time quantitative polymerase chain reaction (qPCR).
Pro A inhibits the proliferation, migration and invasion of Panc-1, BxPC-3 and AsPC-1 PC cells , and Panc-1 cells display the highest sensitivity with an IC50 at the nano-molar level. , Pro A treatment inhibits tumor progression in Panc-1 xenograft nude mice. Pro A treatment promotes both cell apoptosis and autophagy, and Pro A-treated PC cells display characteristics of mitochondrial damage including increased ROS generation, intracellular Ca levels and disruption of MMP. In addition, high sensitivity towards Pro A of Panc-1 cells compared to BxPC-3 and AsPC-1 cells could be partially attributed to the loss of endogenous SMAD4 expression in the latter.
Our findings suggest that Pro A constitutes a promising therapeutic candidate for certain types of PC.
胰腺癌（PC）是一种具有高度转移性和致死性的癌症，总体5年生存率极低。迫切需要为这种致命疾病确定新的治疗药物。强心苷（CGs）传统上因其强大的心血管活性而被使用，最近也有报道称其具有抗肿瘤作用。海葱次苷A（Pro A）是一种天然强心苷，已被证明对多种癌症类型具有抗肿瘤作用。
使用细胞活力测定、集落形成测定和Transwell测定来确定Pro A对PC细胞的细胞毒性作用。通过流式细胞术检测细胞凋亡、细胞周期、活性氧（ROS）生成、细胞内钙水平和线粒体膜电位（MMP）。使用Panc-1异种移植小鼠模型评估Pro A对肿瘤生长的影响。通过透射电子显微镜观察线粒体形态。通过GFP-LC3荧光显微镜评估LC3聚集。通过蛋白质免疫印迹或实时定量聚合酶链反应（qPCR）检测基因表达。
Pro A抑制Panc-1、BxPC-3和AsPC-1 PC细胞的增殖、迁移和侵袭，Panc-1细胞表现出最高的敏感性，IC50处于纳摩尔水平。Pro A治疗可抑制Panc-1异种移植裸鼠的肿瘤进展。Pro A治疗促进细胞凋亡和自噬，经Pro A处理的PC细胞表现出线粒体损伤的特征，包括ROS生成增加、细胞内钙水平升高和MMP破坏。此外，与BxPC-3和AsPC-1细胞相比，Panc-1细胞对Pro A的高敏感性可能部分归因于后者内源性SMAD4表达的缺失。
我们的研究结果表明，Pro A是某些类型胰腺癌的有前景的治疗候选药物。
