Wang Dan, Wang Shasha, Liu Wenjing, Li Minjing, Zheng Qiusheng, Li Defang
Yantai Key Laboratory of Pharmacology of Traditional Chinese Medicine in Tumor Metabolism, School of Integrated Traditional Chinese and Western Medicine, Binzhou Medical University, Yantai, Shandong, P. R. China.
Collaborative Innovation Platform for Modernization and Industrialization of Regional Characteristic Traditional Chinese medicine, School of Integrated Traditional Chinese and Western Medicine, Binzhou Medical University, Yantai, Shandong, P. R. China.
J Pharm Pharmacol. 2022 Aug 9. doi: 10.1093/jpp/rgac044.
Hydroxysafflor yellow B (HSYB) is extracted from the petals of the safflower, a Chinese medicine. Relevant research results have demonstrated that HSYA can suppress the abnormal tumour cell proliferation and induce cell apoptosis. However, the properties of HSYB have rarely been reported, especially its antitumour effects on gastric cancer (GC).
SGC-7901 and BGC-823 cells were treated with different concentrations of HSYB. Cell proliferation inhibition rate was detected by 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay and colony formation detection. The changes in morphology of cells was observed by Hoechst 33258 staining. Cell apoptosis was evaluated by Annexin V-FITC/PI (fluoresceinisothiocyanate/propidium iodide) double staining. JC-1 was used to detect the level of mitochondrial membrane potential (MMP). The protein levels of cleaved-caspase-3, cleaved-caspase-9, APAF-1, cytoplasmic cytochrome C, BAX and BCL-2 were examined by western blot.
HSYB significantly suppressed the proliferation of SGC-7901 and BGC-823 cells. Hoechst 33258 staining assay showed that HSYB treatment triggered apoptotic morphology and the apoptotic rates were significantly increased after being treated with HSYB and the mitochondrial membrane potential was gradually decreased in human GC cells. In addition, Western blot analysis revealed that the levels of cleaved-caspase-3 and cleaved-caspase-9 were remarkably increased in HSYB-treated BGC-823 and SGC-7901 cells. And, the levels of apoptotic protease activating factor-1 (APAF-1) and cytoplasmic cytochrome C were remarkably up-regulated in HSYB-treated cells. At the same time, HSYB could up-regulate the level of BAX and down-regulate the level of BCL-2.
Our data suggest that HSYB could induce GC cell apoptosis via the mitochondrial pathway.
羟基红花黄色素B（HSYB）是从中药红花花瓣中提取的。相关研究结果表明，羟基红花黄色素A（HSYA）可抑制肿瘤细胞异常增殖并诱导细胞凋亡。然而，关于HSYB的性质鲜有报道，尤其是其对胃癌（GC）的抗肿瘤作用。
用不同浓度的HSYB处理SGC-7901和BGC-823细胞。采用3-(4,5-二甲基噻唑-2)-2,5-二苯基四氮唑溴盐（MTT）法和集落形成检测法检测细胞增殖抑制率。通过Hoechst 33258染色观察细胞形态变化。采用膜联蛋白V-异硫氰酸荧光素/碘化丙啶（FITC/PI）双染法评估细胞凋亡。用JC-1检测线粒体膜电位（MMP）水平。通过蛋白质印迹法检测裂解的半胱天冬酶-3、裂解的半胱天冬酶-9、凋亡蛋白酶激活因子-1（APAF-1）、细胞质细胞色素C、Bax和Bcl-2的蛋白水平。
HSYB显著抑制SGC-7901和BGC-823细胞的增殖。Hoechst 33258染色分析表明HSYB处理引发凋亡形态，经HSYB处理后凋亡率显著增加，人胃癌细胞线粒体膜电位逐渐降低。此外，蛋白质印迹分析显示，在经HSYB处理的BGC-823和SGC-7901细胞中，裂解的半胱天冬酶-3和裂解的半胱天冬酶-9水平显著升高。并且，在经HSYB处理的细胞中，凋亡蛋白酶激活因子-1和细胞质细胞色素C水平显著上调。同时，HSYB可上调Bax水平并下调Bcl-2水平。
我们的数据表明HSYB可通过线粒体途径诱导胃癌细胞凋亡。
