INTRODUCTIONThe antitumor activity of norcantharidin (NCTD),the demethylated analogue of cantharidin,wasstudied in the early 1980s in China.NCTD has noside effects on urinary organs which cantharidin hasshown and is e...INTRODUCTIONThe antitumor activity of norcantharidin (NCTD),the demethylated analogue of cantharidin,wasstudied in the early 1980s in China.NCTD has noside effects on urinary organs which cantharidin hasshown and is easier to synthesize,and it can inhibitthe proliferation of several tumor cell lines as wellas transplanted tumors.Clinical trials with NCTD asa monotherapeutic agent indicated that NCTD hadbeneficial effects in patients with different kinds展开更多
BACKGROUND: Gallbladder carcinoma, a lethal malignant neoplasm with poor prognosis, has dismal results of surgical resection and chemoradiotherapy. We previously reported that norcantharidin (NCTD) is useful against g...BACKGROUND: Gallbladder carcinoma, a lethal malignant neoplasm with poor prognosis, has dismal results of surgical resection and chemoradiotherapy. We previously reported that norcantharidin (NCTD) is useful against growth, proliferation, and invasion of human gallbladder carcinoma GBC-SD cells in vitro. In this study, we further studied the inhibitory effect of NCTD on the growth of xenografted tumors of human gallbladder carcinoma in nude mice in vivo and the underlying mechanisms. METHODS: The tumor xenograft model of human gallbladder carcinoma in nude mice in vivo was established with subcutaneous GBC-SD cells. The experimental mice were randomly divided into control, 5-FU, NCTD, and NCTD+5-FU groups which were given different treatments. Tumor growth in terms of size, growth curve, and inhibitory rate was evaluated. Cell cycle, apoptosis, and morphological changes of the xenografted tumors were assessed by flow cytometry and light/electron microscopy. The expression of the cell cycle-related proteins cyclin-D1 and p27 as well as the apoptosis-related proteins Bcl-2, Box, and survivin were determined by the streptavidin-biotin complex (SABC) method and RT-PCR. RESULTS: NCTD inhibited the growth of the xenografted tumors in a dose- and time-dependent manner. Tumor volume decreased (5.61+/-0.39 vs. 9.78+/-0.61 cm(3), P=0.000) with an increased tumor inhibitory rate (42.63% vs. 0%, P=0.012) in the NTCD group compared with the control group. The apoptosis rate increased (15.08+/-1.49% vs. 5.49+/-0.59%, P=0.0001) along with a decreased percentage of cells in S phase (43.47+/-2.83% vs. 69.85+/-1.96%, P=0.0001) in the NTCD group compared with the control group. The morphological changes of apoptosis such as nuclear shrinkage, chromatin aggregation, chromosome condensation, and typical apoptosis bodies in the xenografted tumor cells induced by NCTD were observed by light and electron microscopy. The expression of cyclin-D1, Bcl-2 and survivin proteins/mRNAs decreased significantly, with increased expression of p27 and Bax proteins/mRNAs in the NCTD group compared with the control group. CONCLUSION: NCTD inhibits the growth of xenografted tumors of human gallbladder carcinoma in nude mice by inducing apoptosis and blocking the cell cycle in vivo.展开更多
Objective: To investigate the gene regulation of taxolinduced apoptosis Methods: Northern blot hybridization, enzyme activity assay of S AdoMet synthetase and flow cytometry were performed in the investigation of ex...Objective: To investigate the gene regulation of taxolinduced apoptosis Methods: Northern blot hybridization, enzyme activity assay of S AdoMet synthetase and flow cytometry were performed in the investigation of expression in the mRNA level and biological action of S AdoMet synthetase in taxol induced apoptosis in human breast cancer cell line (BCap 37) Results: Up regulation of S AdoMet synthetase expression was resulted by taxol treatment and the expression peaked at 48 hours Moreover,the up regulation of S AdoMet synthetase was associated with cytotoxicity of anti microtubule agents including taxol and colchicine Inhibition rate of S AdoMet synthetase activity by 1% DMSO was 34% in taxol treated cells and 14% in taxol untreated cells compared to control groups, respectively Posttreatment with 1% DMSO following pretreatment with individual antitumor agent for 3 hrs promoted apoptotic cell death of taxol ,colchicine ,and adriamycin treated Bcap37 cells Conclusion : The induction of apoptosis enhanced by post treatment with DMSO in taxol treated cells is probably linked to its inhibition on enzyme activity of S AdoMet synthetase ,suggesting that the increased expression of S AdoMet synthetase possibly plays an important role in protecting cells from DNA fragmentation in taxol induced apoptosis Accepted July 26, 1998 This work was supported by the National High Biotechnology Foundation of China and a grant from the Natural Science Foundation of Zhejiang Province, China展开更多
Objective: To investigate the mechanism by which taxol induces apoptosis in human breast cancer cells. Methods: Cell morphology, agarose gel electrophoresis, flow cytometry, video time lapse monitor and Western blot...Objective: To investigate the mechanism by which taxol induces apoptosis in human breast cancer cells. Methods: Cell morphology, agarose gel electrophoresis, flow cytometry, video time lapse monitor and Western blot were performed for investigating taxol induced apoptosis in human breast cancer cells (BCap 37). Results: BCap 37 cells treated with taxol (100 nm) under went the arrests of cell mitosis at metaphase of mitosis and induction of apoptosis. Apoptotic cells demonstrated cell shrinkage, condensation or fragmentation of chro mosomes. Nuclear DNA of apoptotic cells displayed ladder bands characteristic of internucleosomal DNA fragmentation. The expression of bcl 2, inhibitor of apotosis, was decreased with modification, while that of bax, inducer of apoptosis, increased only at early stage of the apoptotic pathway and decreased later. Conclusion: In human breast cancer cells the induction of apoptosis by taxol was closely associated with mitotic arrest of cell cycle, and altered expressions of bcl 2 and bax gene possibly played an important role in regulating taxol induced apoptosis.展开更多
Objective: This study was undertaken to determine the expression of apoptosis in breast cancer and to evaluate its significance as a prognostic marker. Methods: A series of 91 invasive breast cancer was analysed for ...Objective: This study was undertaken to determine the expression of apoptosis in breast cancer and to evaluate its significance as a prognostic marker. Methods: A series of 91 invasive breast cancer was analysed for the expression of apoptosis by using the 3 end labeling method of DNA in tissue sections. The apoptotic indexes were the percentages of apoptotic cells among tumor cells. Results: The end labeling method allowed a precise evaluation of the expression of apoptosis. Apoptosis occurred in 91.1% of breast cancer patients, and apoptotic indexes were divided into two groups, 0 0.21 and 0.28 0.62. Low apoptotic index was related to axillary lymph node metastasis (P<0.01). In survival analysis, higher apoptotic index was related to disease free survival (P=0.0095) and overall survival (P=0.0348) in the entire cohort. Cox's analysis showed that apoptotic index had no independent prognostic value. Conclusion: The apoptosis was a spontaneous pheno menon in breast cancer tissue, and the expression was different from each other. Further analysis was needed to clarify the relationship between apoptosis and prognosis, especially the response to adjuvant therapy.展开更多
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, Golgiosome atrophy, mitochondrial swelling, nuclear shrinkage, chromosome condensation and typical apoptosis bodies were seen by electron-microscopy, and the morphological changes of apoptosis occurred in GBCSD cells. The expression of PCNA, Ki-67 and Bcl-2 proteins decreased significantly; the Pix or relative levels of PCNA mRNA, cyclin-D-1 mRNA, Bcl-2 mRNA and Survivin mRNA decreased significantly, whereas the Pix or relative levels of p27 mRNA and Bax mRNA increased significantly. CONCLUSIONS: NCTD inhibits the growth of human gallbladder carcinoma GBC-SD cells in vitro. Its anticancer mechanism may correlate with inhibition of cell proliferation, arrest of the cell cycle, blockage of DNA synthesis, influence on cell metabolism, induction of cell apoptosis and influence on expression of the proliferation-related genes PCNA, Ki-67, cyclin-D-1 and p27, and the apoptosis-related genes Bcl-2, Bax and Survivin in human gallbladder carcinoma GBC-SD cells.展开更多
OBJECTIVE： To investigate the effects of ophiopogonin D on human breast cancer MCF-7 cells METHODS： Cell viability was assessed by 3-（4,5-dimethylthiazol-2-yl）-2,5-diphenyltetrazolium bromide and colony formation ...OBJECTIVE： To investigate the effects of ophiopogonin D on human breast cancer MCF-7 cells METHODS： Cell viability was assessed by 3-（4,5-dimethylthiazol-2-yl）-2,5-diphenyltetrazolium bromide and colony formation experiments. Cell cycle was measured with cell cycle flow cytometry and a living cell assay. Apoptosis and terminal deoxynucleoitidyl transferase-mediated dUTP nick end labeling assays were performed to detect the apoptosis of MCF-7 cells induced by ophiopogonin D. Finally, Western blotting was used to explore the mechanism. RESULTS： Exposure of cells to ophiopogonin D resulted in marked decreases in viable cells and colony formation with a dose-dependent manner. Treatment of these cells with ophiopogonin D also resulted in cell cycle arrest at the G2/M phase, and increased apoptosis. Mechanistically, ophiopogonin D-induced G2/M cell cycle arrest was associated with down-regulation of cyclin BI. Furthermore, activation of caspase-8 and caspase-9 was involved in ophiopogonin D-induced apoptosis. CONCLUSION： The data suggested that ophiopogonin D inhibits MCF-7 cell growth via the induction of cell cycle arrest at the G2/M phase.展开更多
文摘INTRODUCTIONThe antitumor activity of norcantharidin (NCTD),the demethylated analogue of cantharidin,wasstudied in the early 1980s in China.NCTD has noside effects on urinary organs which cantharidin hasshown and is easier to synthesize,and it can inhibitthe proliferation of several tumor cell lines as wellas transplanted tumors.Clinical trials with NCTD asa monotherapeutic agent indicated that NCTD hadbeneficial effects in patients with different kinds
文摘BACKGROUND: Gallbladder carcinoma, a lethal malignant neoplasm with poor prognosis, has dismal results of surgical resection and chemoradiotherapy. We previously reported that norcantharidin (NCTD) is useful against growth, proliferation, and invasion of human gallbladder carcinoma GBC-SD cells in vitro. In this study, we further studied the inhibitory effect of NCTD on the growth of xenografted tumors of human gallbladder carcinoma in nude mice in vivo and the underlying mechanisms. METHODS: The tumor xenograft model of human gallbladder carcinoma in nude mice in vivo was established with subcutaneous GBC-SD cells. The experimental mice were randomly divided into control, 5-FU, NCTD, and NCTD+5-FU groups which were given different treatments. Tumor growth in terms of size, growth curve, and inhibitory rate was evaluated. Cell cycle, apoptosis, and morphological changes of the xenografted tumors were assessed by flow cytometry and light/electron microscopy. The expression of the cell cycle-related proteins cyclin-D1 and p27 as well as the apoptosis-related proteins Bcl-2, Box, and survivin were determined by the streptavidin-biotin complex (SABC) method and RT-PCR. RESULTS: NCTD inhibited the growth of the xenografted tumors in a dose- and time-dependent manner. Tumor volume decreased (5.61+/-0.39 vs. 9.78+/-0.61 cm(3), P=0.000) with an increased tumor inhibitory rate (42.63% vs. 0%, P=0.012) in the NTCD group compared with the control group. The apoptosis rate increased (15.08+/-1.49% vs. 5.49+/-0.59%, P=0.0001) along with a decreased percentage of cells in S phase (43.47+/-2.83% vs. 69.85+/-1.96%, P=0.0001) in the NTCD group compared with the control group. The morphological changes of apoptosis such as nuclear shrinkage, chromatin aggregation, chromosome condensation, and typical apoptosis bodies in the xenografted tumor cells induced by NCTD were observed by light and electron microscopy. The expression of cyclin-D1, Bcl-2 and survivin proteins/mRNAs decreased significantly, with increased expression of p27 and Bax proteins/mRNAs in the NCTD group compared with the control group. CONCLUSION: NCTD inhibits the growth of xenografted tumors of human gallbladder carcinoma in nude mice by inducing apoptosis and blocking the cell cycle in vivo.
文摘Objective: To investigate the gene regulation of taxolinduced apoptosis Methods: Northern blot hybridization, enzyme activity assay of S AdoMet synthetase and flow cytometry were performed in the investigation of expression in the mRNA level and biological action of S AdoMet synthetase in taxol induced apoptosis in human breast cancer cell line (BCap 37) Results: Up regulation of S AdoMet synthetase expression was resulted by taxol treatment and the expression peaked at 48 hours Moreover,the up regulation of S AdoMet synthetase was associated with cytotoxicity of anti microtubule agents including taxol and colchicine Inhibition rate of S AdoMet synthetase activity by 1% DMSO was 34% in taxol treated cells and 14% in taxol untreated cells compared to control groups, respectively Posttreatment with 1% DMSO following pretreatment with individual antitumor agent for 3 hrs promoted apoptotic cell death of taxol ,colchicine ,and adriamycin treated Bcap37 cells Conclusion : The induction of apoptosis enhanced by post treatment with DMSO in taxol treated cells is probably linked to its inhibition on enzyme activity of S AdoMet synthetase ,suggesting that the increased expression of S AdoMet synthetase possibly plays an important role in protecting cells from DNA fragmentation in taxol induced apoptosis Accepted July 26, 1998 This work was supported by the National High Biotechnology Foundation of China and a grant from the Natural Science Foundation of Zhejiang Province, China
文摘Objective: To investigate the mechanism by which taxol induces apoptosis in human breast cancer cells. Methods: Cell morphology, agarose gel electrophoresis, flow cytometry, video time lapse monitor and Western blot were performed for investigating taxol induced apoptosis in human breast cancer cells (BCap 37). Results: BCap 37 cells treated with taxol (100 nm) under went the arrests of cell mitosis at metaphase of mitosis and induction of apoptosis. Apoptotic cells demonstrated cell shrinkage, condensation or fragmentation of chro mosomes. Nuclear DNA of apoptotic cells displayed ladder bands characteristic of internucleosomal DNA fragmentation. The expression of bcl 2, inhibitor of apotosis, was decreased with modification, while that of bax, inducer of apoptosis, increased only at early stage of the apoptotic pathway and decreased later. Conclusion: In human breast cancer cells the induction of apoptosis by taxol was closely associated with mitotic arrest of cell cycle, and altered expressions of bcl 2 and bax gene possibly played an important role in regulating taxol induced apoptosis.
文摘Objective: This study was undertaken to determine the expression of apoptosis in breast cancer and to evaluate its significance as a prognostic marker. Methods: A series of 91 invasive breast cancer was analysed for the expression of apoptosis by using the 3 end labeling method of DNA in tissue sections. The apoptotic indexes were the percentages of apoptotic cells among tumor cells. Results: The end labeling method allowed a precise evaluation of the expression of apoptosis. Apoptosis occurred in 91.1% of breast cancer patients, and apoptotic indexes were divided into two groups, 0 0.21 and 0.28 0.62. Low apoptotic index was related to axillary lymph node metastasis (P<0.01). In survival analysis, higher apoptotic index was related to disease free survival (P=0.0095) and overall survival (P=0.0348) in the entire cohort. Cox's analysis showed that apoptotic index had no independent prognostic value. Conclusion: The apoptosis was a spontaneous pheno menon in breast cancer tissue, and the expression was different from each other. Further analysis was needed to clarify the relationship between apoptosis and prognosis, especially the response to adjuvant therapy.
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, Golgiosome atrophy, mitochondrial swelling, nuclear shrinkage, chromosome condensation and typical apoptosis bodies were seen by electron-microscopy, and the morphological changes of apoptosis occurred in GBCSD cells. The expression of PCNA, Ki-67 and Bcl-2 proteins decreased significantly; the Pix or relative levels of PCNA mRNA, cyclin-D-1 mRNA, Bcl-2 mRNA and Survivin mRNA decreased significantly, whereas the Pix or relative levels of p27 mRNA and Bax mRNA increased significantly. CONCLUSIONS: NCTD inhibits the growth of human gallbladder carcinoma GBC-SD cells in vitro. Its anticancer mechanism may correlate with inhibition of cell proliferation, arrest of the cell cycle, blockage of DNA synthesis, influence on cell metabolism, induction of cell apoptosis and influence on expression of the proliferation-related genes PCNA, Ki-67, cyclin-D-1 and p27, and the apoptosis-related genes Bcl-2, Bax and Survivin in human gallbladder carcinoma GBC-SD cells.
基金Financial assistance was provided by grants from the National Natural Science Foundation of China(No.81173141)
文摘OBJECTIVE： To investigate the effects of ophiopogonin D on human breast cancer MCF-7 cells METHODS： Cell viability was assessed by 3-（4,5-dimethylthiazol-2-yl）-2,5-diphenyltetrazolium bromide and colony formation experiments. Cell cycle was measured with cell cycle flow cytometry and a living cell assay. Apoptosis and terminal deoxynucleoitidyl transferase-mediated dUTP nick end labeling assays were performed to detect the apoptosis of MCF-7 cells induced by ophiopogonin D. Finally, Western blotting was used to explore the mechanism. RESULTS： Exposure of cells to ophiopogonin D resulted in marked decreases in viable cells and colony formation with a dose-dependent manner. Treatment of these cells with ophiopogonin D also resulted in cell cycle arrest at the G2/M phase, and increased apoptosis. Mechanistically, ophiopogonin D-induced G2/M cell cycle arrest was associated with down-regulation of cyclin BI. Furthermore, activation of caspase-8 and caspase-9 was involved in ophiopogonin D-induced apoptosis. CONCLUSION： The data suggested that ophiopogonin D inhibits MCF-7 cell growth via the induction of cell cycle arrest at the G2/M phase.
