BACKGROUND: Gallbladder carcinoma is a lethal malignant neoplasm with dismal surgical results. Unfortunately, the adjuvant therapies for gallbladder carcinoma such as chemotherapy and radiotherapy are also disappointi...BACKGROUND: Gallbladder carcinoma is a lethal malignant neoplasm with dismal surgical results. Unfortunately, the adjuvant therapies for gallbladder carcinoma such as chemotherapy and radiotherapy are also disappointing. We reported that norcantharidin (NCTD), a demethylated form of cantharidin, which is an active ingredient of the Chinese medicine Mylabris, was used against human gallbladder carcinoma GBC-SD cells. In the present study, we further studied the mechanism underlying the inhibitory effect of NCTD on growth of human gallbladder carcinoma GBC-SD cells in vitro. METHODS: Human gallbladder carcinoma GBC-SD cells were grown in cell culture and divided into a NCTD group and a control group. The inhibitory effect of NCTD on growth of GBC-SD cells was investigated by evaluation of proliferation, cell cycle, apoptosis and morphological changes of the cells. Cell proliferation was assessed by tetrazolium-based colorimetric assay. The induction of cell cycle arrest and apoptosis was measured by flow cytometry. The morphological changes of the cells were observed by light- and electron-microscopy. To elucidate the anticancer mechanism of NCTD, expression of the proliferation-related gene proteins PCNA, Ki-67, cyclin-D-1 and p27 and the apoptosis-related gene proteins Bcl-2, Bax and Survivin were determined by the streptavidin-biotin complex method and RT-PCR. RESULTS: NCTD inhibited the proliferation of GBCSD cells in a dose- and time-dependent manner, with an IC50 of 56.18 mu g/ml at 48 hours. The flow cytometric profiles revealed that NCTD (at the IC50 for 48 hours) significantly increased the proportion of cells in G(2)/M phase and significantly decreased the proportion of cells in S phase, with a significantly increased rate of cell apoptosis. After treatment with the 48-hour IC50 dose of NCTD, cell shrinkage, vacuolar cytoplasm, membrane budding, karyorrhexis, karyolysis, chromosome condensation and chromatin aggregation in some GBCSD cells were observed by light-microscopy; decreased microvilli, Golgiosom展开更多
目的:探讨黑蒜提取液(aged black garlic extract,ABGE)对恶性黑色素瘤B16细胞增殖与凋亡影响。方法:MTT法测定黑蒜提取液对黑色素瘤B16细胞株生长抑制率,Hoechst荧光染色观察细胞凋亡及形态的变化,蛋白免疫印迹技术对经不同浓度黑蒜提...目的:探讨黑蒜提取液(aged black garlic extract,ABGE)对恶性黑色素瘤B16细胞增殖与凋亡影响。方法:MTT法测定黑蒜提取液对黑色素瘤B16细胞株生长抑制率,Hoechst荧光染色观察细胞凋亡及形态的变化,蛋白免疫印迹技术对经不同浓度黑蒜提取液处理后的B16细胞中Akt、PTEN、P-Akt蛋白进行检测,Realtime-PCR检测Bcl-2、Bax的表达情况。结果:黑蒜提取液对黑色素瘤B16细胞株有生长抑制作用及促凋亡作用,经不同浓度黑蒜提取液处理后的B16细胞中Akt、P-Akt蛋白的表达明显低于空白对照组,而PTEN的表达则显著上调,Realtime-PCR检测实验各组Bcl-2 mRNA表达均较空白对照组降低,Bax蛋白表达随黑蒜提取液浓度增加逐渐增强。结论:黑蒜提取液可抑制恶性黑色素瘤B16细胞的增殖并诱导其凋亡,其作用机制可能与调控癌基因及凋亡基因表达有关。展开更多
文摘BACKGROUND: Gallbladder carcinoma is a lethal malignant neoplasm with dismal surgical results. Unfortunately, the adjuvant therapies for gallbladder carcinoma such as chemotherapy and radiotherapy are also disappointing. We reported that norcantharidin (NCTD), a demethylated form of cantharidin, which is an active ingredient of the Chinese medicine Mylabris, was used against human gallbladder carcinoma GBC-SD cells. In the present study, we further studied the mechanism underlying the inhibitory effect of NCTD on growth of human gallbladder carcinoma GBC-SD cells in vitro. METHODS: Human gallbladder carcinoma GBC-SD cells were grown in cell culture and divided into a NCTD group and a control group. The inhibitory effect of NCTD on growth of GBC-SD cells was investigated by evaluation of proliferation, cell cycle, apoptosis and morphological changes of the cells. Cell proliferation was assessed by tetrazolium-based colorimetric assay. The induction of cell cycle arrest and apoptosis was measured by flow cytometry. The morphological changes of the cells were observed by light- and electron-microscopy. To elucidate the anticancer mechanism of NCTD, expression of the proliferation-related gene proteins PCNA, Ki-67, cyclin-D-1 and p27 and the apoptosis-related gene proteins Bcl-2, Bax and Survivin were determined by the streptavidin-biotin complex method and RT-PCR. RESULTS: NCTD inhibited the proliferation of GBCSD cells in a dose- and time-dependent manner, with an IC50 of 56.18 mu g/ml at 48 hours. The flow cytometric profiles revealed that NCTD (at the IC50 for 48 hours) significantly increased the proportion of cells in G(2)/M phase and significantly decreased the proportion of cells in S phase, with a significantly increased rate of cell apoptosis. After treatment with the 48-hour IC50 dose of NCTD, cell shrinkage, vacuolar cytoplasm, membrane budding, karyorrhexis, karyolysis, chromosome condensation and chromatin aggregation in some GBCSD cells were observed by light-microscopy; decreased microvilli, Golgiosom
文摘目的:探讨黑蒜提取液(aged black garlic extract,ABGE)对恶性黑色素瘤B16细胞增殖与凋亡影响。方法:MTT法测定黑蒜提取液对黑色素瘤B16细胞株生长抑制率,Hoechst荧光染色观察细胞凋亡及形态的变化,蛋白免疫印迹技术对经不同浓度黑蒜提取液处理后的B16细胞中Akt、PTEN、P-Akt蛋白进行检测,Realtime-PCR检测Bcl-2、Bax的表达情况。结果:黑蒜提取液对黑色素瘤B16细胞株有生长抑制作用及促凋亡作用,经不同浓度黑蒜提取液处理后的B16细胞中Akt、P-Akt蛋白的表达明显低于空白对照组,而PTEN的表达则显著上调,Realtime-PCR检测实验各组Bcl-2 mRNA表达均较空白对照组降低,Bax蛋白表达随黑蒜提取液浓度增加逐渐增强。结论:黑蒜提取液可抑制恶性黑色素瘤B16细胞的增殖并诱导其凋亡,其作用机制可能与调控癌基因及凋亡基因表达有关。
