Antitumor Effects of Ginkgolic Acid in Human Cancer Cell Occur via Cell Cycle Arrest and Decrease the Bcl-2/Bax Ratio to Induce Apoptosis
Antitumor Effects of Ginkgolic Acid in Human Cancer Cell Occur via Cell Cycle Arrest and Decrease the Bcl-2/Bax Ratio to Induce Apoptosis
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DOI：
10.1159/000317750
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发表时间：
2010-10
期刊：
影响因子：
3.3
通讯作者：
Chenchen Zhou;Xiaoyu Li;Wei-Dong Du;Yun Feng;Xiang-li Kong;Yan Li;Li-ying Xiao;Ping Zhang
中科院分区：
文献类型：
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作者：
Chenchen Zhou;Xiaoyu Li;Wei-Dong Du;Yun Feng;Xiang-li Kong;Yan Li;Li-ying Xiao;Ping Zhang
背景：银杏酸（Ginkgolic acids, GAs）是从银杏种皮中提取的，我们之前的研究表明，GA单体可以显著抑制Hep-2的生长，并诱导染色体DNA的断裂。为了进一步评估GA的抗肿瘤潜力，使其成为一种候选的新型抗肿瘤药物，我们对GA的抗肿瘤机制进行了研究。方法：采用MTT比色法测定GA单体对非致瘤性MC-3T3-E1和致瘤性Hep-2、Tac8113细胞株的细胞毒性和抗肿瘤作用。采用MTT比色法和CFSE标记法分析GA单体对肿瘤细胞株增殖的影响。碘化丙啶、膜联蛋白V-FITC染色后，流式细胞术检测细胞周期分布及凋亡细胞百分比。Western blot检测凋亡蛋白Bcl-2、Bax、caspase-3的表达。结果：GA对肿瘤细胞系的抑制作用呈剂量依赖性和时间依赖性。GA作用72 h后，70.53±4.54%的Hep-2和63.5±7.2%的Tca8113细胞阻滞于GO/G1期，凋亡率分别为40.4±1.58和38.4±1.7%。经ga处理的活化caspase-3下调抗凋亡Bcl-2蛋白的表达，上调促凋亡Bax蛋白的表达，最终导致肿瘤细胞中Bcl-2/Bax比值降低。结论：GA的抗肿瘤作用可能是通过抑制细胞分裂、延缓细胞周期进程、诱导细胞凋亡等方式抑制细胞增殖，是抗肿瘤新药的候选药物。
Background: Ginkgolic acids (GAs), extracted from the seed coat of Ginkgo biloba L. Our previous study has shown that GA monomer could inhibit the growth of Hep-2 significantly and induce the fragmentation of the chromosomal DNA. To further assess the antitumor potential and turn it into a candidate new antitumor drug, the antitumor mechanism of GA was investigated. Method: The cytotoxicity and antitumor effect of GA monomer were assayed by MTT colorimetric assay with nontumorogenic MC-3T3-E1 as well as tumorogenic Hep-2 and Tac8113 cell lines. The effect of GA monomer on the proliferation of tumor cell lines was analyzed with MTT colorimetric and CFSE labeled assay. Cell cycle distribution and measurement of the percentage of apoptotic cells were performed by flow cytometry following stained with propidium iodide, annexin V-FITC. The expression of apoptotic proteins Bcl-2, Bax and caspase-3 was analyzed with Western blot. Result: GA only inhibited the growth of tumorogenic cell lines in a both dose- and time-dependent manner. Tumor cells were treated with GA for 72 h, 70.53 ± 4.54% Hep-2 and 63.5 ± 7.2% Tca8113 cells were retarded at GO/G1 phase, and the percentage of apoptosis was 40.4 ± 1.58 and 38.4 ± 1.7%, respectively. GA-treated activated caspase-3 downregulated the expression of anti-apoptotic Bcl-2 protein and upregulated the expression of pro-apoptotic Bax protein, eventually leading to a decrease in the Bcl-2/Bax ratio in tumor cells. Conclusions: The antitumor action of GA was due to inhibiting the proliferation in a manner of inhibiting division, retarding the progress of cell cycle and inducing apoptosis, making GA a candidate as new antitumor drug.