Tan Jun, Tian Ying, Cai Renlian, Yi Tianci, Jin Daochao, Guo Jianjun
Provincial Key Laboratory for Agricultural Pest Management of Mountainous Region, Institute of Entomology, Guizhou University, Guiyang, Guizhou 550025, China.
Department of Histology and Embryology, Zunyi Medical University, Zunyi, Guizhou 563000, China.
Evid Based Complement Alternat Med. 2019 Jan 3;2019:8934794. doi: 10.1155/2019/8934794. eCollection 2019.
Dallas is used as a traditional Chinese medicine. In China, clinical evidence suggests that it has anticancer activity. However, the anticancer active components are not fully elucidated. In the present study, we purified an anticancer active component (named CHP) from To gain a comprehensive insight into the protein components, shotgun proteomic analysis was conducted. The anticancer active protein band was cut from the sodium dodecyl sulphate-polyacrylamide gel electrophoresis gel and digested with trypsin to generate peptide mixture. The peptide fragments were then analysed by liquid chromatography tandem mass spectrometry; 18 proteins were identified. In addition, we evaluated the effects of CHP on the proliferation and apoptosis of two human gastric cancer cell lines (SGC-7901 and BGC-823). The cultured cells were treated with CHP at concentrations of 20, 30, and 40 g/mL. Inhibition of cell growth was determined by the MTT assay. Hoechst 33258 staining was adopted to detect apoptosis morphologically. Apoptotic cells were quantified by Annexin V-FITC/propidium iodide staining and flow cytometry. Tumour growth was assessed by subcutaneous inoculation of 4T1 cells into BALB/c mice. There was a concentration- and time-dependent decrease in the proliferation of both cell lines at CHP concentrations of 20-40 g/mL. Apoptotic characteristics, such as karyopyknotic pyknic hyperfluorescence bolus and nuclear fragmentation, were observed in both the cell lines by Hoechst 33258 staining. Flow cytometry showed that CHP induced significant ( < 0.01) concentration-dependent apoptosis of SGC-7901 cells. assay showed that CHP can partially inhibit tumour growth derived from 4T1 cells . The present study is the first to report that CHP in inhibits the proliferation of cancer cell lines via the suppression of cancer cell proliferation and acceleration of apoptosis.
漏芦被用作一种传统中药。在中国,临床证据表明它具有抗癌活性。然而,其抗癌活性成分尚未完全阐明。在本研究中,我们从……中纯化出一种抗癌活性成分(命名为CHP)。为了全面了解蛋白质成分,进行了鸟枪法蛋白质组学分析。从十二烷基硫酸钠 - 聚丙烯酰胺凝胶电泳凝胶上切下抗癌活性蛋白条带,并用胰蛋白酶消化以产生肽混合物。然后通过液相色谱串联质谱分析肽片段;鉴定出了18种蛋白质。此外,我们评估了CHP对两种人胃癌细胞系(SGC - 7901和BGC - 823)增殖和凋亡的影响。将培养的细胞用浓度为20、30和40μg/mL的CHP处理。通过MTT法测定细胞生长抑制情况。采用Hoechst 33258染色从形态学上检测凋亡。通过膜联蛋白V - FITC/碘化丙啶染色和流式细胞术对凋亡细胞进行定量。通过将4T1细胞皮下接种到BALB/c小鼠中来评估肿瘤生长。在CHP浓度为20 - 40μg/mL时,两种细胞系的增殖均呈现浓度和时间依赖性下降。通过Hoechst 33258染色在两种细胞系中均观察到凋亡特征,如核固缩、核致密高荧光团块和核碎裂。流式细胞术显示CHP诱导SGC - 7901细胞发生显著(P < 0.01)的浓度依赖性凋亡。……实验表明CHP可部分抑制源自4T1细胞的肿瘤生长。本研究首次报道漏芦中的CHP通过抑制癌细胞增殖和加速凋亡来抑制癌细胞系的增殖。