20(S)-人参皂苷Rg3诱导肺癌细胞NCI-H460凋亡的机制

doi:吉林省科技厅科技发展计划项目资助课题 (20

20(S)-人参皂苷Rg3诱导肺癌细胞NCI-H460凋亡的机制

王光兰¹，崔俊生²，王心蕊¹，石博¹，高静¹，倪劲松¹

（1.吉林大学基础医学院病理生物学教育部重点实验室，吉林长春 130021；2.吉林大学中日联谊医院病理科，吉林长春 130033）

收稿日期:2008-03-18 修回日期:1900-01-01 出版日期:2008-11-28 发布日期:2008-11-28
通讯作者: 倪劲松

Mechanism of apoptosis of lung carcinoma cell line NCI-H460 induced by 20(S)-ginsenoside Rg3

WANG Guang-lan¹，CUI Jun-sheng²，WANG Xin-rui¹，SHI Bo¹，GAO Jing¹，NI Jin-Song¹

(1.Key Laboratory of Pathobiology,Ministry of Education,School of Basic Medical Sciences,Jilin University,Changchun 130021,China; 2.Department of Pathology,China-Japan Union Hospital,Jilin University,Changchun 130033,China)

Received:2008-03-18 Revised:1900-01-01 Online:2008-11-28 Published:2008-11-28
Contact: NI Jin-Song

摘要/Abstract

摘要： 目的：探讨20(S)-人参皂苷Rg3(SPG-Rg3)诱导人大细胞肺癌细胞NCI-H460凋亡及其机制。方法：MTT法测定NCI-H460细胞的增殖，依据其半数抑制浓度（IC₅₀）将SPG-Rg3实验组分为25、50及100 mg•L^-1组，并设空白对照组。采用AO/EB荧光双染、免疫细胞化学染色、Rhodamine 123摄取法及Fluo-3/AM染色分别观察NCI-H460细胞的凋亡、Bcl-2及Bax蛋白的表达量、线粒体跨膜电位及细胞内游离钙离子浓度。结果：SPG-Rg3能抑制NCI-H460的生长，抑制率与浓度呈正相关 (r=0.764，P<0.05)，其IC₅₀值为47.97 mg•L^-1；SPG-Rg3组NCI-H460细胞呈现明显凋亡改变;SPG-Rg3 100 mg•L^-1组细胞内Bcl-2蛋白的表达量明显低于空白对照组（P<0.01）；SPG-Rg3各浓度组细胞Bax蛋白的表达量均明显高于对照组（P<0.001），并随浓度的增大而增加（P<0.01）；各浓度组细胞内Bax/Bcl-2比值均高于对照组，细胞内线粒体跨膜电位则均低于对照组(P<0.01)，并随药物作用浓度的增加而减少，组间比较差异有显著性(P<0.01)；各组细胞内游离钙离子浓度明显高于对照组 (P<0.01)。结论：SPG-Rg3能够明显抑制细胞的增殖，诱导细胞凋亡。其作用机制可能是SPG-Rg3促进细胞内Bax蛋白的表达并增加Bax/Bcl-2比值，降低线粒体跨膜电位，提高细胞内游离钙离子浓度，最终经线粒体通路诱导细胞凋亡。

关键词: 肺肿瘤, 细胞凋亡, 钙离子, 线粒体跨膜电位, 凋亡相关因子

Abstract: To investigate the mechanism of apoptosis of lung carcinoma cell line NCI-H460 induced by 20(S)-ginsenoside Rg3 (SPG-Rg3).Methods MTT assay was used to detect the proliferation rate of NCI-H460 cells.The cells were treated with SPG-Rg3 and divided into three groups based on IC₅₀ value: SPG-Rg3 25 mg•L^-1 group，SPG-Rg3 50 mg•L^-1 group and SPG-Rg3 100 mg•L^-1 group.Control group was also set up.Then the AO/EB fluorescence double-dye technique,immunocytochemistry staining,Fluo-3 staining and Rhodamine 123 uptake assay approaches were adopted to observe the apoptotic rate,expression levels of Bcl-2 and Bax proteins,concentration of cytoplasmic free Ca²⁺ and mitochondrial transmembrane potential of NCI-H460 cells. Results SPG-Rg3 obviously inhibited the cell proliferation,showing a positive correlation between inhibitory effect and concentration (r=0.764,P<0.05)，and the IC₅₀ was 47.97 mg•L^-1.There were visible apoptotic changes observed in SPG-Rg3 groups.The protein expression level of Bcl-2 in SPG-Rg3 100 mg•L^-1 group was less than that in control group(P<0.01).The protein expression levels of Bax in SPG-Rg3 groups were more than that in control group（P<0.001）,and they were positively correlated with the drug dose（P<0.01）.The ratios of Bax to Bcl-2 in SPG-Rg3 groups were bigger than that in control group, while the mitochondrial transmembrane potential showed an opposite situation(P<0.01) and it decreased when the drug dose increased.There were significantly differences between various groups (P<0.01).The concentrations of cytoplasmic free Ca²⁺ in SPG-Rg3 groups were higher than that in control group(P<0.01). Conclusion SPG-Rg3 could obviously inhibit the cell proliferation and induce the apoptosis.The mechanism is concerned with that SPG-Rg3 could up-regulate the expression level of Bax protein and increase the ratio of Bax to Bcl-2,down-regulate the mitochondrial transmembrane potential and increase the cytoplasmic free Ca²⁺ concentration,lead to the induction of apoptosis via mitochondrial pathway.

Key words: lung carcinoma, apoptosis, Ca^2+<, sup>, mitochondrial transmembrane potential, apoptosis related factor

中图分类号:

R-332

王光兰，崔俊生，王心蕊，石博，高静，倪劲松. 20(S)-人参皂苷Rg3诱导肺癌细胞NCI-H460凋亡的机制[J]. J4, 2008, 34(6): 980-984.

WANG Guang-lan，CUI Jun-sheng，WANG Xin-rui，SHI Bo，GAO Jing，NI Jin-Song. Mechanism of apoptosis of lung carcinoma cell line NCI-H460 induced by 20(S)-ginsenoside Rg3[J]. J4, 2008, 34(6): 980-984.

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