Shen Huamei, Zhao Shuai, Xu Zhipeng, Zhu Lihua, Han Yuehua, Ye Jun
Department of Infectious Disease, The Second Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou, Zhejiang 310009, P.R. China.
Department of Gastroenterology, The Second Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou, Zhejiang 310009, P.R. China.
Oncol Lett. 2015 Jul;10(1):367-371. doi: 10.3892/ol.2015.3153. Epub 2015 Apr 27.
In the present study, the effects of evodiamine on the apoptosis of human gastric cancer cells was studied in order to assess its antitumor effects and identify the molecular mechanisms involved. SGC7901 gastric cancer cells were treated with evodiamine at various concentrations (0, 3, 6, 12, 24 and 48 µmol/l) for 24 h. Inhibition of the proliferation of SGC7901 cells was assessed using an MTT assay. The morphology of treated SGC7901 cells was observed using optical microscopy; in addition, the effect of evodiamine on the nuclear morphology of cells was analyzed using Hoechst 33258 staining with fluorescence microscopy. Annexin V-fluorescein isothiocyanate/propidium iodide staining and flow cytometric analysis were used for investigating the effect of evodiamine on the induction of apoptosis in SGC7901 cells. Expression levels of survivin and caspase-3 were examined using reverse transcription polymerase chain reaction. The results demonstrated that evodiamine significantly inhibited SGC7901 cell proliferation (P<0.05) and induced apoptosis in a dose-dependent manner (P<0.05). Morphological characteristics of apoptosis were confirmed using optical microscopy and Hoechst 33258 staining analysis indicated that evodiamine treatment resulted in the typical characteristics of apoptotic programmed cell death, including cell shrinkage and apoptotic body formation. Flow cytometric analysis demonstrated that evodiamine induced the dose-dependent apoptosis of SGC7901 cells. Messenger (m)RNA levels of survivin decreased and those of caspase-3 increase in a dose-dependent manner in SGC7901 cells treated with various concentrations of evodiamine for 24 h. In conclusion, the results of the present study demonstrated that evodiamine inhibited proliferation and induced apoptosis in gastric cancer cells via the downregulation of survivin and upregulation of caspase-3 mRNA.
在本研究中,为了评估吴茱萸碱的抗肿瘤作用并确定其相关分子机制,研究了吴茱萸碱对人胃癌细胞凋亡的影响。用不同浓度(0、3、6、12、24和48 μmol/l)的吴茱萸碱处理SGC7901胃癌细胞24小时。采用MTT法评估SGC7901细胞增殖的抑制情况。用光学显微镜观察经处理的SGC7901细胞的形态;此外,用荧光显微镜对经Hoechst 33258染色的细胞进行分析,以研究吴茱萸碱对细胞核形态的影响。采用膜联蛋白V-异硫氰酸荧光素/碘化丙啶染色及流式细胞术分析,研究吴茱萸碱对SGC7901细胞凋亡诱导的影响。采用逆转录聚合酶链反应检测生存素和半胱天冬酶-3的表达水平。结果表明,吴茱萸碱显著抑制SGC7901细胞增殖(P<0.05),并以剂量依赖性方式诱导细胞凋亡(P<0.05)。光学显微镜观察证实了凋亡的形态学特征,Hoechst 33258染色分析表明,吴茱萸碱处理导致凋亡程序性细胞死亡的典型特征,包括细胞收缩和凋亡小体形成。流式细胞术分析表明,吴茱萸碱诱导SGC7901细胞呈剂量依赖性凋亡。在经不同浓度吴茱萸碱处理24小时的SGC7901细胞中,生存素的信使核糖核酸(mRNA)水平呈剂量依赖性下降,而半胱天冬酶-3的mRNA水平呈剂量依赖性升高。总之,本研究结果表明,吴茱萸碱通过下调生存素和上调半胱天冬酶-3 mRNA抑制胃癌细胞增殖并诱导其凋亡。