Wu Jin-Yi, Sheikho Almutamad, Ma He, Li Tian-Ci, Zhao Ya-Qi, Zhang Ya-Lin, Wang Dun
Institute of Entomology, Northwest A&F University, P.O. Box 712100, Yangling, Shaanxi, China.
Department of Basic Sciences, Faculty of Allied Medical Sciences, University of Nyala, Nyala, South Darfur, Sudan.
Cytotechnology. 2017 Dec;69(6):875-883. doi: 10.1007/s10616-017-0102-7. Epub 2017 May 24.
Bombyx batryticatus is a traditional Chinese medicine. To understand apoptotic effect of B. batryticatus ethanol extract (BBE), we investigated the role of BBE in inducing apoptosis of human gastric cancer cells SGC-7901. Cells treated with BBE and apoptosis was assessed by methyl thiazolyl tetrazolium (MTT) assay, morphological changes, DNA fragmentation and flow cytometry assays. The expression of Bcl-2, Bax and P21 were evaluated by western blot analysis and real time polymerase chain reaction. MTT assay showed that the cytotoxicity of BBE extract on SGC-7901 cells was correlated with treatment time and concentration. After treatment with 6 mg/mL of BBE the microscopy showed that, the majority of SGC-7901 cells were obviously reduced, distorted and grew slowly. Annexin-V/propidium iodide double-staining assay emerge the early apoptosis and the late apoptosis after treatment with different times by laser confocal fluorescence microscopy and flow cytometer. Cell cycle analysis of SGC 79 cells showed that BBE induced cell cycle arrest in the G1 and G2 phases. DNA fragmentation indicated the trend of BBE inducing apoptosis on SGC-7901 cells. The qRT-PCR and western blot analysis indicated that the mRNA and protein expressions of Bax and P21 were significantly up-regulated whereas that of Bc1-2 was down-regulated after treatment with BBE for 24 h. Our results revealed a correlation between gene regulation and BBE-induced apoptosis, which might indicate the potential of BBE in cancer therapy.
僵蚕是一种传统中药。为了解僵蚕乙醇提取物（BBE）的凋亡作用，我们研究了BBE在诱导人胃癌细胞SGC - 7901凋亡中的作用。用BBE处理细胞，并通过甲基噻唑基四氮唑（MTT）法、形态学变化、DNA片段化和流式细胞术分析来评估细胞凋亡情况。通过蛋白质免疫印迹分析和实时聚合酶链反应评估Bcl - 2、Bax和P21的表达。MTT法显示，BBE提取物对SGC - 7901细胞的细胞毒性与处理时间和浓度相关。用6mg/mL的BBE处理后，显微镜观察显示，大多数SGC - 7901细胞明显减少、变形且生长缓慢。通过激光共聚焦荧光显微镜和流式细胞仪对不同处理时间后的细胞进行膜联蛋白V/碘化丙啶双染分析，发现了早期凋亡和晚期凋亡情况。SGC 79细胞的细胞周期分析表明，BBE诱导细胞周期停滞在G1期和G2期。DNA片段化显示了BBE诱导SGC - 7901细胞凋亡的趋势。定量逆转录聚合酶链反应（qRT - PCR）和蛋白质免疫印迹分析表明，用BBE处理24小时后，Bax和P21的mRNA和蛋白质表达显著上调，而Bcl - 2的表达下调。我们的结果揭示了基因调控与BBE诱导的凋亡之间的相关性，这可能表明BBE在癌症治疗中的潜力。
