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
AIM:To investigate the inhibitory effect of acetylshikonin on human gastric carcinoma cell line SGC-7901 and its mechanism. METHODS:MTT assay was used to assess the inhibitory effect of acetylshikonin on proliferation...AIM:To investigate the inhibitory effect of acetylshikonin on human gastric carcinoma cell line SGC-7901 and its mechanism. METHODS:MTT assay was used to assess the inhibitory effect of acetylshikonin on proliferation of SGC-7901 cells.Apopt osis-inducing effect was determined by flow cytometry and terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end-labeling with Hoechst staining.Expression of mRNA and protein in Bcl-2 and Bax was analyzed by reverse transcription-polymerase chain reaction and Western blot.Antitumor effect of acetylshikonin on a mouse SGC-7901 model was also determined. RESULTS:Forty-eight hours after treatment with acetylshikonin,MTT assay showed that acetylshikonin inhibited the proliferation of SGC-7901 cells in a dose-dependent manner.The half maximal inhibitory concentration of acetylshikonin to SGC-7901 cells was 0.428±0.07 mg/L.Cell shrinkage,nuclear pyknosis and chromatin condensation,which are the characteristics of cell apoptosis,were observed in treated SGC-7901 cells and the percentage of apoptosis increased in a dose-dependent manner.Acetylshikonin downregulated the expression of Bcl-2 and up-regulated the expression of Bax in the treated SGC-7901 cells compared with the controls.The experiment in vivo showed that 0.5,1,and 2 mg/kg of acetylshikonin significantly inhibited the growth of tumor in the mouse SGC-7901 model,with an inhibitory rate of 25.00%-55.76%. CONCLUSION:Acetylshikonin inhibits the growth of SGC-7901 cells in vitro and in vivo by inducing cell apoptosis.展开更多
Objective： The aim of the study was to construct miRNA-451 expression vector pLMP-miRNA-451 which could help identify the functions of miRNA-451 in SGC-7901 cell. Methods： Total RNA was extracted from SGC-7901 cells...Objective： The aim of the study was to construct miRNA-451 expression vector pLMP-miRNA-451 which could help identify the functions of miRNA-451 in SGC-7901 cell. Methods： Total RNA was extracted from SGC-7901 cells to synthesized cDNA. The synthesized cDNA encoding pre-miRNA-451 was amplified by polymerase chain reaction （PCR）. The PCR product was separated by electrophoresis on 1% agarose gel and then recovered and purified. The purified cDNA fragments of miRNA-451 precursor sequence was then ligated with vector pLMP for 1 h by using DNA ligase to form pLMP- miRNA-451 plasmid. After that, the pLMP-miRNA-451 plasmid was transformed into E. coli DH5a strain expression system to clone and amplificate. The purified pLMP-miRNA-451 extracted from E. coli DH5a via transformation and clone screening was identificatied with restriction enzyme digestion and DNA sequencing. At last, pLMP-miRNA-451 was transfected into SGC-7901 cells with lip2000. Real-time PCR was used for detection of the miRNA-451, the transfection efficiency was ob- served under fluorescence microscopy and cell counting kit-8 assay was conduced to evaluate the effect of miRNA-451 on SGC-7901 cell proliferation. Results： Our results showed that pLMP-miRNA-451 expression vector was not only constructed successfully and effectively infected SGC-7901 cells, but also could repress the SGC-7901 cell proliferation. Conclusion： The constructed plasmid pLMP-miRNA-451 could used for further studies of miRNA-451 in SGC-7901 cell lines.展开更多
[Objectives]To observe the effect of drug-containing serum from different extracts of Phyllanthus reticulatus leaf on the proliferation of human gastric cancer SGC-7901 cells and the effect of in vivo on the life exte...[Objectives]To observe the effect of drug-containing serum from different extracts of Phyllanthus reticulatus leaf on the proliferation of human gastric cancer SGC-7901 cells and the effect of in vivo on the life extension rate of mice with H22 ascites tumor,and to investigate the effects of chemical components such as Corilagin,ethyl gallate,ethyl brevifolincarboxylate,and gallic acid on the proliferation of human gastric cancer SGC-7901 cells.[Methods]MTT method was used to observe the inhibitory effect of drug-containing serum of different extracts of P.reticulatus leaf on human gastric cancer SGC-7901 cells in vitro to determine its active site.The active site was used as the research object to establish a Kunming mouse ascites tumor model,to investigate the effect on the life extension rate of mice with H22 ascites tumor,to further separate the monomer components from the effective fraction and investigate its effect on human gastric cancer SGC-7901 cells.[Results]Compared with the 10%blank serum control group,10%drug-containing serum of ethyl acetate and n-butanol in P.reticulatus leaf had significant inhibitory effect on the proliferation of human gastric cancer SGC-7901 cells,and the inhibition rate was 34.99%and 28.68%,respectively(P<0.05).Compared with the blank group,the survival time of the high dose group of ethyl acetate in P.reticulatus leaf was significant(P<0.05).Gallic acid,Corilagin,and Brevifolincarboxylic acid ethyl ester had a significant inhibitory effect on the proliferation of human gastric cancer SGC-7901 cells,with IC 50 of 26.52,70.45,and 158.86μg/mL,respectively.Ethyl gallate had no inhibitory effect on the proliferation of human gastric cancer SGC-7901 cells,and its IC 50 was 251.96μg/mL.[Conclusions]The drug-containing serum of ethyl acetate and n-butanol extract of P.reticulatus can inhibit the proliferation of human gastric cancer SGC-7901 cells.Ethyl acetate of P.reticulatus leaves can increase the life extension rate of mice with H22 ascites tumor.Gallic acid,Corilagin,and Brevifolincarboxylic acid ethyl ester isolated from its active site are the material basis for inhibiting the proliferation of human gastric cancer SGC-7901 cells.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
Objective: To research the inhibitory effect on SGC-7901 cells of α-pinene, and the related mechanism of α-pinene. Methods: Used the MTT method to detect inhibition rate and western blotting to detect the influence ...Objective: To research the inhibitory effect on SGC-7901 cells of α-pinene, and the related mechanism of α-pinene. Methods: Used the MTT method to detect inhibition rate and western blotting to detect the influence on expression of ATM, Phos-S1981ATM, H2AX, γH2AX, CHK2 and p-CHK2, p53 and phos-p53 cell cycle related protein in SGC-7901 cells. Results: The research found α-pinene could inhibit the proliferation of SGC-7901 cells observably in vitro, and the inhibition rate assumes the dependence on concentration;and western blotting results showed that, α-pinene could activate phospho-ATM, increase the amount of γH2AX (p p < 0.05);increase the expression of p-CHK2, p53 and phos-p53 (p p p < 0.05);But there is no significant effect on expression of CHK2 (p > 0.05). Conclusions: α-pinene could inhibit the proliferation of SGC-7901 cells, and by inducing ATM (Ataxia Telangiectasia-Mutated) kinase signal pathway in DNA damage response, activating cell cycle checkpoint, making the cell cycle arrest then exerts its anti-tumor effects.展开更多
目的通过测定隐丹参酮对人胃癌SGC-7901细胞增殖及血管内皮生长因子（VEGF）mRNA的影响,探讨其抗肿瘤的作用机制。方法采用四甲基偶氮唑蓝（MTT）比色法检测隐丹参酮对人胃癌SGC-7901细胞增殖的影响,采用Real Time RT-PCR检测隐丹参酮对...目的通过测定隐丹参酮对人胃癌SGC-7901细胞增殖及血管内皮生长因子（VEGF）mRNA的影响,探讨其抗肿瘤的作用机制。方法采用四甲基偶氮唑蓝（MTT）比色法检测隐丹参酮对人胃癌SGC-7901细胞增殖的影响,采用Real Time RT-PCR检测隐丹参酮对VEGF m RNA表达的影响。结果 20、40、60、80、100μmol/L的隐丹参酮作用于人胃癌SGC-7901细胞6-48 h,其生长和增殖均受到一定程度的抑制;24 h为隐丹参酮对SGC-7901细胞抑制作用的最佳时间,IC50为59.11μmol/L;选取40、60和80μmol/L 3个剂量作用于人胃癌细胞SGC7901 24 h后,60和80μmol/L的隐丹参酮均可下调VEGF m RNA的表达（均P〈0.01）。结论隐丹参酮可抑制人胃癌SGC-7901细胞增殖,并能抑制VEGF mRNA的表达,提示这可能是其抗肿瘤的作用机制之一。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.
文摘AIM:To investigate the inhibitory effect of acetylshikonin on human gastric carcinoma cell line SGC-7901 and its mechanism. METHODS:MTT assay was used to assess the inhibitory effect of acetylshikonin on proliferation of SGC-7901 cells.Apopt osis-inducing effect was determined by flow cytometry and terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end-labeling with Hoechst staining.Expression of mRNA and protein in Bcl-2 and Bax was analyzed by reverse transcription-polymerase chain reaction and Western blot.Antitumor effect of acetylshikonin on a mouse SGC-7901 model was also determined. RESULTS:Forty-eight hours after treatment with acetylshikonin,MTT assay showed that acetylshikonin inhibited the proliferation of SGC-7901 cells in a dose-dependent manner.The half maximal inhibitory concentration of acetylshikonin to SGC-7901 cells was 0.428±0.07 mg/L.Cell shrinkage,nuclear pyknosis and chromatin condensation,which are the characteristics of cell apoptosis,were observed in treated SGC-7901 cells and the percentage of apoptosis increased in a dose-dependent manner.Acetylshikonin downregulated the expression of Bcl-2 and up-regulated the expression of Bax in the treated SGC-7901 cells compared with the controls.The experiment in vivo showed that 0.5,1,and 2 mg/kg of acetylshikonin significantly inhibited the growth of tumor in the mouse SGC-7901 model,with an inhibitory rate of 25.00%-55.76%. CONCLUSION:Acetylshikonin inhibits the growth of SGC-7901 cells in vitro and in vivo by inducing cell apoptosis.
基金supported by a grant from the Natural Science Foundation of China (No. 8100098)
文摘Objective： The aim of the study was to construct miRNA-451 expression vector pLMP-miRNA-451 which could help identify the functions of miRNA-451 in SGC-7901 cell. Methods： Total RNA was extracted from SGC-7901 cells to synthesized cDNA. The synthesized cDNA encoding pre-miRNA-451 was amplified by polymerase chain reaction （PCR）. The PCR product was separated by electrophoresis on 1% agarose gel and then recovered and purified. The purified cDNA fragments of miRNA-451 precursor sequence was then ligated with vector pLMP for 1 h by using DNA ligase to form pLMP- miRNA-451 plasmid. After that, the pLMP-miRNA-451 plasmid was transformed into E. coli DH5a strain expression system to clone and amplificate. The purified pLMP-miRNA-451 extracted from E. coli DH5a via transformation and clone screening was identificatied with restriction enzyme digestion and DNA sequencing. At last, pLMP-miRNA-451 was transfected into SGC-7901 cells with lip2000. Real-time PCR was used for detection of the miRNA-451, the transfection efficiency was ob- served under fluorescence microscopy and cell counting kit-8 assay was conduced to evaluate the effect of miRNA-451 on SGC-7901 cell proliferation. Results： Our results showed that pLMP-miRNA-451 expression vector was not only constructed successfully and effectively infected SGC-7901 cells, but also could repress the SGC-7901 cell proliferation. Conclusion： The constructed plasmid pLMP-miRNA-451 could used for further studies of miRNA-451 in SGC-7901 cell lines.
基金Supported by Natural Science Research Project of Guangxi University of Chinese Medicine in 2015"Study on Anti-tumor Effect of Phyllanthus reticulatus Leaf Extract"(p15028)Independent Research Program of Guangxi Key Laboratory of Traditional Chinese Medicine Pharmacology in 2015"Study on Phyllanthus reticulatus Leaf 7404 Nude Mice Xenografts"+1 种基金Young Teachers Enhancement Project of Guangxi Department of Education in 2018"Study on the Mechanism of Zhuang Medicine Phyllanthus reticulatus Leaf against Liver Cancer"(2018KY0300)2019-2021 Guangxi First-class Discipline Construction Open Project Fund for Young Scholars of Guangxi University of Chinese Medicine"Anti-hepatocarcinoma Mechanism of Autophagy Induced by Ethyl Acetate in Phyllanthus reticulatus Leaf Based on PI3K/AKT and STAT3/m TOR Signaling Pathway"(2019XK089)
文摘[Objectives]To observe the effect of drug-containing serum from different extracts of Phyllanthus reticulatus leaf on the proliferation of human gastric cancer SGC-7901 cells and the effect of in vivo on the life extension rate of mice with H22 ascites tumor,and to investigate the effects of chemical components such as Corilagin,ethyl gallate,ethyl brevifolincarboxylate,and gallic acid on the proliferation of human gastric cancer SGC-7901 cells.[Methods]MTT method was used to observe the inhibitory effect of drug-containing serum of different extracts of P.reticulatus leaf on human gastric cancer SGC-7901 cells in vitro to determine its active site.The active site was used as the research object to establish a Kunming mouse ascites tumor model,to investigate the effect on the life extension rate of mice with H22 ascites tumor,to further separate the monomer components from the effective fraction and investigate its effect on human gastric cancer SGC-7901 cells.[Results]Compared with the 10%blank serum control group,10%drug-containing serum of ethyl acetate and n-butanol in P.reticulatus leaf had significant inhibitory effect on the proliferation of human gastric cancer SGC-7901 cells,and the inhibition rate was 34.99%and 28.68%,respectively(P<0.05).Compared with the blank group,the survival time of the high dose group of ethyl acetate in P.reticulatus leaf was significant(P<0.05).Gallic acid,Corilagin,and Brevifolincarboxylic acid ethyl ester had a significant inhibitory effect on the proliferation of human gastric cancer SGC-7901 cells,with IC 50 of 26.52,70.45,and 158.86μg/mL,respectively.Ethyl gallate had no inhibitory effect on the proliferation of human gastric cancer SGC-7901 cells,and its IC 50 was 251.96μg/mL.[Conclusions]The drug-containing serum of ethyl acetate and n-butanol extract of P.reticulatus can inhibit the proliferation of human gastric cancer SGC-7901 cells.Ethyl acetate of P.reticulatus leaves can increase the life extension rate of mice with H22 ascites tumor.Gallic acid,Corilagin,and Brevifolincarboxylic acid ethyl ester isolated from its active site are the material basis for inhibiting the proliferation of human gastric cancer SGC-7901 cells.
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Objective: To research the inhibitory effect on SGC-7901 cells of α-pinene, and the related mechanism of α-pinene. Methods: Used the MTT method to detect inhibition rate and western blotting to detect the influence on expression of ATM, Phos-S1981ATM, H2AX, γH2AX, CHK2 and p-CHK2, p53 and phos-p53 cell cycle related protein in SGC-7901 cells. Results: The research found α-pinene could inhibit the proliferation of SGC-7901 cells observably in vitro, and the inhibition rate assumes the dependence on concentration;and western blotting results showed that, α-pinene could activate phospho-ATM, increase the amount of γH2AX (p p < 0.05);increase the expression of p-CHK2, p53 and phos-p53 (p p p < 0.05);But there is no significant effect on expression of CHK2 (p > 0.05). Conclusions: α-pinene could inhibit the proliferation of SGC-7901 cells, and by inducing ATM (Ataxia Telangiectasia-Mutated) kinase signal pathway in DNA damage response, activating cell cycle checkpoint, making the cell cycle arrest then exerts its anti-tumor effects.
文摘目的通过测定隐丹参酮对人胃癌SGC-7901细胞增殖及血管内皮生长因子（VEGF）mRNA的影响,探讨其抗肿瘤的作用机制。方法采用四甲基偶氮唑蓝（MTT）比色法检测隐丹参酮对人胃癌SGC-7901细胞增殖的影响,采用Real Time RT-PCR检测隐丹参酮对VEGF m RNA表达的影响。结果 20、40、60、80、100μmol/L的隐丹参酮作用于人胃癌SGC-7901细胞6-48 h,其生长和增殖均受到一定程度的抑制;24 h为隐丹参酮对SGC-7901细胞抑制作用的最佳时间,IC50为59.11μmol/L;选取40、60和80μmol/L 3个剂量作用于人胃癌细胞SGC7901 24 h后,60和80μmol/L的隐丹参酮均可下调VEGF m RNA的表达（均P〈0.01）。结论隐丹参酮可抑制人胃癌SGC-7901细胞增殖,并能抑制VEGF mRNA的表达,提示这可能是其抗肿瘤的作用机制之一。
