波罗蜜属药用植物化合物对SMMC-7721和SGC-7901细胞凋亡的影响

Effects of cycloartocarpin A and artocarpin extracted from Fructus Artocarpi Heterophylli on apoptosis of SMMC-7721 and SGC-7901 cells

目的:研究波罗蜜属药用植物单体化合物(ACR-2、ACR-3)对人肝癌细胞(SMMC-7721)和胃腺癌细胞(SGC-7901)凋亡的影响。方法:SMMC-7721和SGC-7901细胞常规培养并加入处理因素,设实验组和阴性对照组,实验组分别加入不同浓度的ACR-2或ACR-3200μL。ACR-2处理SMMC-7721细胞的终浓度分别是3.46×10-3、13.82×10-3、55.30×10-3mmol/L,ACR-3处理SMMC-7721细胞的终浓度分别是6.88×10-3、27.52×10-3、110.09×10-3mmol/L;ACR-2处理SGC-7901细胞的终浓度分别是8.06×10-3、32.26×10-3、129.03×10-3mmol/L,ACR-3处理SGC-7901细胞的终浓度分别是2.87×10-3、11.47×10-3、45.87×10-3mmol/L。阴性对照组加入200μLPBS。培养48h。分别采用Hoechst33342/碘化丙锭(propidiumiodide,PI)双标记荧光染色法、原位末端标记(terminaldeoxynucleotidyltransferase-mediateddeoxyuridinetriphosphate-biotinnickendlabeling,TUNEL)法和流式细胞术,检测ACR-2、ACR-3对SMMC-7721、SGC-7901细胞凋亡的影响。结果:Hoechst33342/PI染色结果显示,化合物ACR-2、ACR-3分别作用于SMMC-7721、SGC-7901细胞后,部分细胞呈凋亡特征性改变,核呈分叶或碎片状,边集,细胞核呈亮蓝色染色。TUNEL结果显示,化合物ACR-2、ACR-3组SMMC-7721、SGC-7901细胞部分凋亡,细胞核为分叶或碎片状,边集,细胞核呈棕色染色。阴性对照组细胞形态呈圆形,核呈蓝色染色,凋亡细胞较少。化合物ACR-2、ACR-3组SMMC-7721细胞和SGC-7901细胞凋亡率较对照组明显升高(P<0.05,P<0.01)。结论:波罗蜜属药用植物单体化合物ACR-2、ACR-3对人肝癌细胞SMMC-7721、胃腺癌细胞SGC-7901具有明显的诱导凋亡作用。

Objective： To investigate the effects of cycloartocarpin A （ACR-2） and artocarpin （ACR-3）, monomeric compounds isolated from Fructus Artocarpi Heterophylli, on apoptosis of SMMC-7721 and SGC-7901 cell lines.Methods： SMMC-7721 and SGC-7901 cells were routinely cultured, and divided into experiment group and control group. The SMMC-7721 cells were treated with different concentrations of ACR-2 （3. 46 × 10^-3, 13.82×10^-3, 55.30×10^-3 mmol/L） and ACR-3 （6.88×10^-3, 27.52×10^- 3 ,110.09X10 3 mmol/L）, and the SGC-7901 cells were also treated with different concentrations of ACR-2 （8.06 ×10^-3, 32.26 × 10 3, 129.03×10^-3 mmol/L） andACR-3 （2.87×10^-3, 11.47×10^-3 ,45.87×10^-3 mmol/L）, with PBS （DMSO〈 0.1%） as control treatment. Cell apoptosis was measured by double labeled staining with Hoechst33342/ propidium iodide （PI） and TdT-mediated dUTP-biotin nick end labeling （TUNEL） and flow cytometry.Results： ACR-2 and ACR-3 could induce apoptosis of SMMC-7721 and SGC-7901 cells. Some of SMMC-7721 and SGC-7901 cells demonstrated typical apoptosis after being treated with ACR-2 and ACR-3. Hoechst33342/PI staining showed that ceils were fraught with overlapping nuclei and nuclear debris or Iobule, and the nuclear appeared light blue. TUNEL showed that cells permeated with overlapping nuclei and nuclear debris or Iobule, and the nuclear appeared brown. Less apoptotic cells were observed in negative control group, and the nuclear appeared light blue. The apoptosis rates of SMMC-7721 and SGC-7901 cells in the ACR-2 and ACR-3 treated groups were significant higher than those in the control group （P〈0.05, P〈0.01）.