Wang Hai-Bing, Ma Xiao-Qiong
National Clinical Research Base of Traditional Chinese Medicine, the First Affiliated Hospital, Zhejiang Chinese Medical University, Hangzhou 310006, China.
Yao Xue Xue Bao. 2012 Jun;47(6):816-21.
This study is to investigate the effect of (2-methyl-n-butyl) shikonin (MBS) on inducing apoptosis of human gastric cancer cell line SGC-7901 and the role of ERK1/2 signal pathway in the apoptosis. MTT assay was used to detect SGC-7901 cell proliferation. DNA condensation was measured by DAPI stain. Cell apoptosis was analyzed by flow cytometry. Mitochondrial membrane potential (MMP) was analyzed by JC-1 staining. The protein expressions of Bcl-2, Bax, Survivin, cleaved caspase-9, cleaved caspase-3, cleaved PARP, p-ERK1/2, ERK1/2, p-JNK, JNK, p-p38 and p38 were detected by Western blotting. The results showed that MBS reduced the cell viability of SGC-7901 cells in a dose- and time-dependent manner. The IC50 at 24 h and 48 h for SGC-7901 cells was 10.113 and 4.196 micromolL(-1), respectively. After being treated with MBS, the typical nuclear condensation was observed in SGC-7901 cells by DAPI stain. Apoptosis in SGC-7901 cells was induced by MBS in a dose dependent manner. The protein expression of Bcl-2 was down-regulated, while the protein expressions of cleaved caspase-9, cleaved caspase-3, cleaved PARP, p-ERK1/2 and p-JNK were up-regulated after MBS treatment. U0126, a specific MAP kinase (MEK1/2) inhibitor, blocked the ERK1/2 activation by MBS. MMP was decreased by MBS treatment. It can be concluded that MBS could inhibit SGC-7901 cell proliferation and induce apoptosis. Mitochondrial apoptosis pathway, ERK1/2 signal pathway and JNK signal pathway might be involved in this process.
本研究旨在探讨（2-甲基-n-丁基）紫草素（MBS）诱导人胃癌细胞系SGC-7901凋亡的作用以及ERK1/2信号通路在该凋亡过程中的作用。采用MTT法检测SGC-7901细胞增殖。通过DAPI染色测定DNA凝聚。采用流式细胞术分析细胞凋亡。通过JC-1染色分析线粒体膜电位（MMP）。采用蛋白质印迹法检测Bcl-2、Bax、Survivin、裂解的caspase-9、裂解的caspase-3、裂解的PARP、p-ERK1/2、ERK1/2、p-JNK、JNK、p-p38和p38的蛋白表达。结果表明，MBS以剂量和时间依赖性方式降低SGC-7901细胞的活力。SGC-7901细胞在24小时和48小时的IC50分别为10.113和4.196微摩尔/升（-1）。用MBS处理后，通过DAPI染色在SGC-7901细胞中观察到典型的核凝聚。MBS以剂量依赖性方式诱导SGC-7901细胞凋亡。MBS处理后，Bcl-2的蛋白表达下调，而裂解的caspase-9、裂解的caspase-3、裂解的PARP、p-ERK1/2和p-JNK的蛋白表达上调。特异性丝裂原活化蛋白激酶（MEK1/2）抑制剂U0126可阻断MBS对ERK1/2的激活。MBS处理可降低MMP。可以得出结论，MBS可抑制SGC-7901细胞增殖并诱导凋亡。线粒体凋亡途径、ERK1/2信号通路和JNK信号通路可能参与这一过程。
