OBJECTIVE To investigate whether nimesulide can suppress tumor growth and induce apoptosis in SGC-7901 gastric cancer cells and to explore the molecular mechanism involved. METHODS SGC-7901 cells were cultured in RPMI...OBJECTIVE To investigate whether nimesulide can suppress tumor growth and induce apoptosis in SGC-7901 gastric cancer cells and to explore the molecular mechanism involved. METHODS SGC-7901 cells were cultured in RPMI 1640 medium containing different concentrations of nimesulide （0,12.5, 50, 100, 200, 400 μmol/L）. The MTT assay, morphological observation, electron microscopy （EM）, immunohistochemical analysis and Western blot analysis were employed to investigate the effects of nimesulide on the SGC-7901 cells and to explore possible related molecular mechanisms. RESULTS Nimesulide inhibited the growth of SGC-7901 cells and elicited typical apoptotic morphologic changes. Nimesulide also decreased NF-κB and Bcl-2 expression, but increased the level of the Bax protein. The positive rate of Bcl-2 protein expression at 0, 50, 100 and 200 μmol/L of nimesulide was 58.3±14.0%, 50.2±9.9%, 32.8±5.0% and 22.7±5.5% respectively based on immunohistochemical staining. The positive rate of Bax protein expression was 22.0±5.7%, 29.2±6.5%, 42.7±5.9% and 74.5±9.1% and the NF-κB expression was 74.2±10.9%, 61.8±7.6%, 36.7±10.9% and 17.5±12.3%, Significant differences were found between so μmol/L and 100 μmol/L and 200μmol/L. Western blot analysis also showed that the expression of NF-κB was decreased. CONCLUSION Nimesulide suppresses tumor growth and induces apoptosis by inhibiting NF-κB expression, which may be related to the overexpression of Bax relative to Bcl-2 expression.展开更多
Objective: To investigate the cytotoxic effects and the potential mechanisms of crebanine N-oxide in SGC-7901 gastric adenocarcinoma cells. Methods: The cytotoxicity of crebanine N-oxide was evaluated by 3-(4,5-dimeth...Objective: To investigate the cytotoxic effects and the potential mechanisms of crebanine N-oxide in SGC-7901 gastric adenocarcinoma cells. Methods: The cytotoxicity of crebanine N-oxide was evaluated by 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide assay and cellular morphology was observed under a microscope. Cell apoptosis was determined by flow cytometry using propidium iodide staining. The expression levels of apoptotic-related proteins, cleaved caspase-3, cytochrome C, p53 and Bax, and autophagyrelated proteins p62, beclin1 and LC3 were detected by Western blotting assays. Results: Crebanine N-oxide treatment significantly inhibited the proliferation of SGC-7901 cells in a dose-dependent and timedependent manner via induction of G2-phase cell cycle arrest, apoptosis, and autophagy in SGC-7901 cells.Conclusions: Crebanine N-oxide could inhibit the growth of gastric cancer cells by promoting apoptosis and autophagy and could be used as a potential agent for treating gastric cancer.展开更多
AIM： To detect the biological characters of the SGC7901 gastric cancer cell-dendritic cell fusion vaccines.METHODS： The suspending living SGC7901 gastric cancer cells and dendritic cells were induced to be fusioned ...AIM： To detect the biological characters of the SGC7901 gastric cancer cell-dendritic cell fusion vaccines.METHODS： The suspending living SGC7901 gastric cancer cells and dendritic cells were induced to be fusioned by polyethylene glycol. Pure fusion cells were obtained by selective culture with the HAT/HT culture systems. The fusion cells were counted at different time points of culture and their growth curves were drawn to reflect their proliferative activities. The fusion cells were also cultured in culture medium to investigate whether they could grow into cell clones. MTT method was used to test the stimulating abilities of the fusion cells on T lymphocytes＇ proliferations. Moreover, the fusion cells were planted into nude mice to observe whether they could grow into new planted tumors in this kind of immunodeficiency animals.RESULTS： The fusion cells had weaker proliferative activity and clone abilities than their parental cells. When they were cultured, the counts of cells did not increase remarkably, nor could they grow into cell clones in culture medium. The fusion cells could not grow into new planted tumors after planted into nude mice. The stimulating abilities of the fusion cells on T lymphocytes＇ proliferations were remarkably increased than their parental dendritic cells. CONCLUSION： The SGC7901 gastric cancer cell-dendritic cell fusion vaccines have much weaker proliferative abilities than their parental cells, but they keep strong abilities to irritate the T lymphocytes and have no abilities to grow into new planted tumors in immunodeficiency animals. These are the biological basis for their antitumor biotherapies.展开更多
Objective:To investigate the effect of Chinese medicine Compound Weichang'an(胃肠安)for invig-orating the spleen on apoptosis of gastric cancer SGC7901 cells and its possible mechanism.Methods:The gas-trie cancer ...Objective:To investigate the effect of Chinese medicine Compound Weichang'an(胃肠安)for invig-orating the spleen on apoptosis of gastric cancer SGC7901 cells and its possible mechanism.Methods:The gas-trie cancer SGC-7901 cells were divided into different mass concentration groups(0 mg·L^(-1),500 mg·L^(-1)1000 mg·L^(-1),1500 mg·L^(-1),2000 mg·L^(-1)).CCK8 and monoclonal test were applied to detect prolifera-tion ability;comet assay was used to detect DNA damage.After DCFH-DA fluorescent labeling,the level of ROS activity was detected by flow cytometer;after AnnexinV-FTC/PI double labeling,the proportion of apoptotic ellls was detected by flow cytometer;after JC-1 staining,the mi tochondri almembrane potential was detected by flow cytometer;after FTTC-DEVD-FMK staining,the ratio of Caspase activity was detected by flow cytometer.Results:Weichang an inhibited cell proliferation and reduced cell colony formation in a time-dose-dependent manner;the results of comet electrophoresis showed that Weichang'an could induce DNA damage in gastric cancer cells;com-pared with control group.the ratio of Weichang'an's intervention with the apoptosis of gastric cancer cells in-creased(P<0.05),the mitochondrial membrane potential decreased(P<0.05),the activity of Caspase3 and Caspase9 increased(P<0.05),and the intracellular ROS level increased(P<0.05).Among them,the effect of Weichang'an treatment group(1000 mg·L^(-1))was the most significant.Conclusion:Weichang'an has an inhibi-tory effect on the proliferation of gastric cancer SGC7901 cells and can induce cell apoptosis.Its mechanism may be related with the ROS-mediated pathway of mitochondrial apoptosis and DNA damage.展开更多
AIM： To explore the effect of trichostatin A （TSA） on apoptosis and acetylated histone H3 levels in gastric cancer cell lines BGC-823 and SGC-7901. METHODS： The effect of TSA on growth inhibition and apoptosis was...AIM： To explore the effect of trichostatin A （TSA） on apoptosis and acetylated histone H3 levels in gastric cancer cell lines BGC-823 and SGC-7901. METHODS： The effect of TSA on growth inhibition and apoptosis was examined by MTT, fluorescence microscopy and PI single-labeled flow cytometry. The acetylated histone H3 level was detected by Western blot. RESULTS： TSA induced apoptosis in gastric cancer cell lines BGC-823 and SGC-7901 was in a dose and time-dependent manner. Apoptotic cells varied significantly between TSA treated groups （37.5 ng/mL 72 h for BGC-823 cell line and 75 ng/mL 72 h for SGC-7901 cell line） and control group （0.85 ± 0.14 vs 1.14 ± 0.07, P = 0.02; 0.94 ± 0.07 vs 1.15 ± 0.06, P = 0.02）. Morphologic changes of apoptosis, including nuclear chromatin condensation and fluorescence strength, were observed under fluorescence microscopy. TSA treatment in BGC-823 and SGC-7901 cell lines obviously induced cell apoptosis, which was demonstrated by the increased percentage of sub-G1 phase cells, the reduction of Gl-phase cells and the increase of apoptosis rates in flow cytometric analysis. The result of Western blot showed that the expression of acetylated histone H3 increased in BGC-823 and SGC-7901 TSA treatment groups as compared with the control group.CONCLUSION： TSA can induce cell apoptosis in BGC-823 and SGC-7901 cell lines. The expression of acetylated histone H3 might be correlated with apoptosis.展开更多
AIM: To investigate the growth effects of 4-phenyl butyric acid (PBA) on human gastric carcinoma cells and their mechanisms. METHODS: Moderately-differentiated human gastric carcinoma SGC-7901 and lowly-differentiated...AIM: To investigate the growth effects of 4-phenyl butyric acid (PBA) on human gastric carcinoma cells and their mechanisms. METHODS: Moderately-differentiated human gastric carcinoma SGC-7901 and lowly-differentiated MGC-803 cells were treated with 5, 10, 20, 40, and 60 μmol/L PBA for 1-4 d. Cell proliferation was detected using the MTT colorimetric assay. Cell cycle distributions were examined using flow cytometry.RESULTS: The proliferation of gastric carcinoma cells was inhibited by PBA in a doseand time-dependent fashion. Flow cytometry showed that SGC-7901 cells treated with low concentrations of PBA were arrested at the G0/G1 phase, whereas cells treated with high concentrations of PBA were arrested at the G2/M phase. Although MGC-803 cells treated with low concentrations of PBA were also arrested at the G0/G1 phase, cells treated with high concentrations of PBA were arrested at the S phase. CONCLUSION: The growth inhibitory effect of PBA on gastric cancer cells is associated with alteration of the cell cycle. For moderately-differentiated gastric cancer cells, the cell cycle was arrested at the G0/G1 and G2/M phases. For lowly-differentiated gastric cancer cells, the cell cycle was arrested at the G0/G1 and S phases.展开更多
基金This work was supported by the grant form the Natural Science Foundation of the Education Department of Jiangsu Province of China (No. 05KJD320234 and 01KJB320011).
文摘OBJECTIVE To investigate whether nimesulide can suppress tumor growth and induce apoptosis in SGC-7901 gastric cancer cells and to explore the molecular mechanism involved. METHODS SGC-7901 cells were cultured in RPMI 1640 medium containing different concentrations of nimesulide （0,12.5, 50, 100, 200, 400 μmol/L）. The MTT assay, morphological observation, electron microscopy （EM）, immunohistochemical analysis and Western blot analysis were employed to investigate the effects of nimesulide on the SGC-7901 cells and to explore possible related molecular mechanisms. RESULTS Nimesulide inhibited the growth of SGC-7901 cells and elicited typical apoptotic morphologic changes. Nimesulide also decreased NF-κB and Bcl-2 expression, but increased the level of the Bax protein. The positive rate of Bcl-2 protein expression at 0, 50, 100 and 200 μmol/L of nimesulide was 58.3±14.0%, 50.2±9.9%, 32.8±5.0% and 22.7±5.5% respectively based on immunohistochemical staining. The positive rate of Bax protein expression was 22.0±5.7%, 29.2±6.5%, 42.7±5.9% and 74.5±9.1% and the NF-κB expression was 74.2±10.9%, 61.8±7.6%, 36.7±10.9% and 17.5±12.3%, Significant differences were found between so μmol/L and 100 μmol/L and 200μmol/L. Western blot analysis also showed that the expression of NF-κB was decreased. CONCLUSION Nimesulide suppresses tumor growth and induces apoptosis by inhibiting NF-κB expression, which may be related to the overexpression of Bax relative to Bcl-2 expression.
基金supported by the National Natural Science Foundation of China(NO.81760628).
文摘Objective: To investigate the cytotoxic effects and the potential mechanisms of crebanine N-oxide in SGC-7901 gastric adenocarcinoma cells. Methods: The cytotoxicity of crebanine N-oxide was evaluated by 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide assay and cellular morphology was observed under a microscope. Cell apoptosis was determined by flow cytometry using propidium iodide staining. The expression levels of apoptotic-related proteins, cleaved caspase-3, cytochrome C, p53 and Bax, and autophagyrelated proteins p62, beclin1 and LC3 were detected by Western blotting assays. Results: Crebanine N-oxide treatment significantly inhibited the proliferation of SGC-7901 cells in a dose-dependent and timedependent manner via induction of G2-phase cell cycle arrest, apoptosis, and autophagy in SGC-7901 cells.Conclusions: Crebanine N-oxide could inhibit the growth of gastric cancer cells by promoting apoptosis and autophagy and could be used as a potential agent for treating gastric cancer.
文摘AIM： To detect the biological characters of the SGC7901 gastric cancer cell-dendritic cell fusion vaccines.METHODS： The suspending living SGC7901 gastric cancer cells and dendritic cells were induced to be fusioned by polyethylene glycol. Pure fusion cells were obtained by selective culture with the HAT/HT culture systems. The fusion cells were counted at different time points of culture and their growth curves were drawn to reflect their proliferative activities. The fusion cells were also cultured in culture medium to investigate whether they could grow into cell clones. MTT method was used to test the stimulating abilities of the fusion cells on T lymphocytes＇ proliferations. Moreover, the fusion cells were planted into nude mice to observe whether they could grow into new planted tumors in this kind of immunodeficiency animals.RESULTS： The fusion cells had weaker proliferative activity and clone abilities than their parental cells. When they were cultured, the counts of cells did not increase remarkably, nor could they grow into cell clones in culture medium. The fusion cells could not grow into new planted tumors after planted into nude mice. The stimulating abilities of the fusion cells on T lymphocytes＇ proliferations were remarkably increased than their parental dendritic cells. CONCLUSION： The SGC7901 gastric cancer cell-dendritic cell fusion vaccines have much weaker proliferative abilities than their parental cells, but they keep strong abilities to irritate the T lymphocytes and have no abilities to grow into new planted tumors in immunodeficiency animals. These are the biological basis for their antitumor biotherapies.
基金We thank for the funding support from the Scientific Research Project of National TCM Clinical Research Base Business Construction of National Administration of Traditional Chi-nese Medicine(JDZX2015068)Henan Sci-ence and Technology Project(202102310164)+1 种基金Henan Scientific Research Project of Traditional Chinese Medicine(2019JDZX025)Scientific Research Project of Henan Province Hospital of TCM(2018YJKT09).
文摘Objective:To investigate the effect of Chinese medicine Compound Weichang'an(胃肠安)for invig-orating the spleen on apoptosis of gastric cancer SGC7901 cells and its possible mechanism.Methods:The gas-trie cancer SGC-7901 cells were divided into different mass concentration groups(0 mg·L^(-1),500 mg·L^(-1)1000 mg·L^(-1),1500 mg·L^(-1),2000 mg·L^(-1)).CCK8 and monoclonal test were applied to detect prolifera-tion ability;comet assay was used to detect DNA damage.After DCFH-DA fluorescent labeling,the level of ROS activity was detected by flow cytometer;after AnnexinV-FTC/PI double labeling,the proportion of apoptotic ellls was detected by flow cytometer;after JC-1 staining,the mi tochondri almembrane potential was detected by flow cytometer;after FTTC-DEVD-FMK staining,the ratio of Caspase activity was detected by flow cytometer.Results:Weichang an inhibited cell proliferation and reduced cell colony formation in a time-dose-dependent manner;the results of comet electrophoresis showed that Weichang'an could induce DNA damage in gastric cancer cells;com-pared with control group.the ratio of Weichang'an's intervention with the apoptosis of gastric cancer cells in-creased(P<0.05),the mitochondrial membrane potential decreased(P<0.05),the activity of Caspase3 and Caspase9 increased(P<0.05),and the intracellular ROS level increased(P<0.05).Among them,the effect of Weichang'an treatment group(1000 mg·L^(-1))was the most significant.Conclusion:Weichang'an has an inhibi-tory effect on the proliferation of gastric cancer SGC7901 cells and can induce cell apoptosis.Its mechanism may be related with the ROS-mediated pathway of mitochondrial apoptosis and DNA damage.
文摘AIM： To explore the effect of trichostatin A （TSA） on apoptosis and acetylated histone H3 levels in gastric cancer cell lines BGC-823 and SGC-7901. METHODS： The effect of TSA on growth inhibition and apoptosis was examined by MTT, fluorescence microscopy and PI single-labeled flow cytometry. The acetylated histone H3 level was detected by Western blot. RESULTS： TSA induced apoptosis in gastric cancer cell lines BGC-823 and SGC-7901 was in a dose and time-dependent manner. Apoptotic cells varied significantly between TSA treated groups （37.5 ng/mL 72 h for BGC-823 cell line and 75 ng/mL 72 h for SGC-7901 cell line） and control group （0.85 ± 0.14 vs 1.14 ± 0.07, P = 0.02; 0.94 ± 0.07 vs 1.15 ± 0.06, P = 0.02）. Morphologic changes of apoptosis, including nuclear chromatin condensation and fluorescence strength, were observed under fluorescence microscopy. TSA treatment in BGC-823 and SGC-7901 cell lines obviously induced cell apoptosis, which was demonstrated by the increased percentage of sub-G1 phase cells, the reduction of Gl-phase cells and the increase of apoptosis rates in flow cytometric analysis. The result of Western blot showed that the expression of acetylated histone H3 increased in BGC-823 and SGC-7901 TSA treatment groups as compared with the control group.CONCLUSION： TSA can induce cell apoptosis in BGC-823 and SGC-7901 cell lines. The expression of acetylated histone H3 might be correlated with apoptosis.
基金Supported by Natural Science Foundation of Ningbo, No. 2009A610134Natural Sciences Foundation of Zhejiang, No. Y207244+3 种基金College Students’ Science-Technology Innovation Program of Zhejiang Province, No. 200959the Excellent Disser-tation Foundation of Ningbo University, No. 201014KC Wong Magna Fund of Ningbo Universitythe Scientific Innovation Team Project of Ningbo, No.2011B82014
文摘AIM: To investigate the growth effects of 4-phenyl butyric acid (PBA) on human gastric carcinoma cells and their mechanisms. METHODS: Moderately-differentiated human gastric carcinoma SGC-7901 and lowly-differentiated MGC-803 cells were treated with 5, 10, 20, 40, and 60 μmol/L PBA for 1-4 d. Cell proliferation was detected using the MTT colorimetric assay. Cell cycle distributions were examined using flow cytometry.RESULTS: The proliferation of gastric carcinoma cells was inhibited by PBA in a doseand time-dependent fashion. Flow cytometry showed that SGC-7901 cells treated with low concentrations of PBA were arrested at the G0/G1 phase, whereas cells treated with high concentrations of PBA were arrested at the G2/M phase. Although MGC-803 cells treated with low concentrations of PBA were also arrested at the G0/G1 phase, cells treated with high concentrations of PBA were arrested at the S phase. CONCLUSION: The growth inhibitory effect of PBA on gastric cancer cells is associated with alteration of the cell cycle. For moderately-differentiated gastric cancer cells, the cell cycle was arrested at the G0/G1 and G2/M phases. For lowly-differentiated gastric cancer cells, the cell cycle was arrested at the G0/G1 and S phases.
