【摘要】 目的:探讨20(S)-人参皂苷Rg3(SPG-Rg3)对人乳腺癌MCF-7细胞的诱导凋亡作用及其可能机制。方法:人乳腺癌细胞MCF-7细胞株分为SPG-Rg3实验组及空白对照组。采用MTT法观察SPG-Rg3对MCF-7细胞生长的抑制作用,并计算出IC50,依据IC50值确定SPG-Rg3的有效浓度;流式细胞术检测SPG-Rg3作用后MCF-7细胞周期的变化;AO/EB双染从形态学上观察SPG-Rg3对MCF-7细胞凋亡的作用;免疫细胞化学染色和RT-PCR技术分别从蛋白水平和分子水平上检测SPG-Rg3对MCF-7细胞的诱导凋亡作用及其与caspase-8基因的关系。结果:SPG-Rg3IC50为(155.70±0.71)mg·L-1,当SPG-Rg3浓度在37.5~600.0mg·L-1时,MCF-7细胞的生长抑制率随SPG-Rg3浓度的增加而增大,与对照组比较,细胞增殖受到明显抑制(P<0.05);MCF-7细胞的生长周期也发生变化,S期细胞数增加,明显高于对照组(P<0.01),G2/M期细胞数则明显少于对照组(P<0.01);并且在G1峰前出现明显的凋亡峰,凋亡细胞数增加。形态学上观察到SPG-Rg3(150mg·L-1)实验组细胞大部分核着黄色荧光或橘红色荧光,形态呈固缩状、圆珠状或新月形,呈现明显的凋亡改变;caspase-8免疫细胞化学染色可见,对照组MCF-7细胞caspase-8蛋白不表达,SPG-Rg3实验组则呈强表达,两组细胞HSCORE得分为1.894±0.027及2.869±0.043,两组比较差异有显著性(P<0.01);提取细胞mRNA并设计caspase-8引物进行RT-PCR,SPG-Rg3实验组细胞caspase-8大量表达,对照组细胞无表达。结论:SPG-Rg3能诱导乳腺癌MCF-7细胞发生凋亡,其机制可能与其活化caspase-8作用有关。更多还原

【Abstract】 Objective To study the effect of 20(S)-ginsendoside Rg3(SPG-Rg3) on apoptosis of human mammary carcinoma line MCF-7 cells and its possible mechanism. Methods Human mammary carcinoma line MCF-7 cells were divided into experiment group of SPG-Rg3 and control group. The inhibition of SPG-Rg3 on the growth of MCF-7 cells was detected by MTT assay,IC50 was calculated to obtain the effective concentration; flow cytometry was used to observe the cell cycle of MCF-7 cells; AO/EB fluorescence double-dye technology was performed to observe the apoptosis of cells in morphology,immunocellularchemistry and RT-PCR were utilized to investigate the apoptosis of MCF-7 cells and its relationship with caspase-8 from protein level and molecule level. Results The IC50 of SPG-Rg3 was (155.7±0.71) mg·L-1,when the concentration of SPG-Rg3 was arranged from 37.5 to 600.0 mg·L-1,the growth inhibitory rate of MCF-7 cells was higher following the increase of its concentration,the cell growth was greatly inhibited compared with control group(P<0.05); cell cycle was changed,the cell number of S period increased compared with control group(P<0.01),the cell number of G2/M period decreased compared with control group(P<0.01),in experiment group there was an apoptotic apex before G1 apex,and the number of apoptotic cells increased; most of cell nuleus in SPG-Rg3 (150 mg·L-1)experiment group appeared yellow or chrysoidine fluorescence,contracted,beaded and crescent; the result of immunocellularchemical staining of caspase-8 indicated that caspase-8 protein had no expression in control group,but stronger expression in SPG-Rg3 group. HSCORE scores of the two groups were 1.894±0.027 and 2.869±0.043,the difference was markable(P<0.01). MCF-7 cells mRNA was extraited and caspase-8 primers were designed to perform RT-PCR,caspase-8 expressed strongly in experiment group and nothing in control. Conclusion SPG-Rg3 could induce MCF-7 cell apoptosis,and the possible mechanism may be related to activating caspase-8.更多还原