Objective： To investigate the As4S4 induced growth inhibition and apoptosis in HeLa cells and its possible relationship with cyclooxygenase-2 （COX-2）. Methods： HeLa cells were treated with various concentrations ...Objective： To investigate the As4S4 induced growth inhibition and apoptosis in HeLa cells and its possible relationship with cyclooxygenase-2 （COX-2）. Methods： HeLa cells were treated with various concentrations （7.5, 15, 30, 60 mg/L） of As4S4 at different times （12, 24, 36, 48, 60 h）. Cell growth was measured by MTT. Apoptosis was detected by double staining flow cytometry （FCM）. Levels of PGE2 were measured by radioimmunoassay. The expression of COX-2 protein was examined by Western blot analysis. Results： After treated with different concentrations of As4S4, the growth of HeLa cells was suppressed significantly in a dose-and time-dependent manner. The IC50 of 24 h was 30 mg/L （P〈0.01）. As4S4 induced apoptosis with apoptosis rates at 8.13%-62.36% by flow cytometry （FCM） in a dose-dependent manners. The release of PGE2 was reduced in HeLa cells with the values being （70.56±2.03）, （48.58±2.28）, （29.25±1.57） and （18.02±1.04） respectively, significantly different compared with control group （3.15±0.01） （P〈0.01）. As4S4 also inhibited the activity and expression of COX-2 in a dose dependent manner and down-regulated the expression of COX-2 protein greatly. Conclusion： As4S4 could inhibit the proliferation and increase apoptosis in human HeLa cells. These effects may depend on the inhibition of the expression of COX-2 and PGE2 by As4S4.展开更多
Mineral medicine,especially those containing heavy metals,is one of the characteristics of traditional Chinese medicine.A famous mineral medicine,realgar,containing heavy metal arsenic with a chemical formula of As4S4...Mineral medicine,especially those containing heavy metals,is one of the characteristics of traditional Chinese medicine.A famous mineral medicine,realgar,containing heavy metal arsenic with a chemical formula of As4S4,has the function of detoxification,killing bacteria and viruses,and eliminating dampness and phlegm.Different As4S4 isomers are likely to have different drug effects and pharmacological actions.Therefore,it is of great scientific significance to find more stable As4S4 isomers.In view of this,ab initio molecular orbital theory and density functional theory(DFT)have been used to study ten isomers of As4S4 at the B3LYP/6-31 G*,B3LYP/6-311+G*,B3LYP/6-311+G(3 df,2 p)and MP2/(6-311+G*,LanL2 MB)levels of theory.In addition to the two isomers having been studied previously,eight new isomers were investigated in the present paper.All the ten As4S4 isomers were proved to be true local minima on their potential energy surfaces.The calculated NICS values and molecular orbital analyses showed that,the D2d symmetric As4S4,isomer 1,may be s-aromatic.The study proves that ten As4S4 isomers are stable thermodynamically,and are highly desirable for the future theoretical study of realgar.展开更多
Objective: To investigate the effect of arsenic sulfide (tetra-arsenic tetra-sulfide As4S4; diarsenic trisulfide As2S3) on tissue factor (TF) expression and procoagulant activity (PCA) of acute promyelocytic leukemia(...Objective: To investigate the effect of arsenic sulfide (tetra-arsenic tetra-sulfide As4S4; diarsenic trisulfide As2S3) on tissue factor (TF) expression and procoagulant activity (PCA) of acute promyelocytic leukemia( APL) cell lines ( NB4 and MR2) and the basic mechanism of their role. Methods: NB4 and MR2 cells were respectively treated with As4S4 , As2S3, As4S4 and Cyclohexamide( CHX). PCA of the cells was detected using one-stage clotting assay. TF antigen was detected by ELISA. TF and PML/RARa fusion gene mRNA by semi-quantitive RT-PCR. The PCA and TF antigen of HL-60 and K562 cells were also examined. Results: The PCA and TF antigen level in NB4 and MR2 cells were significantly higher than that in HL-60 and K562 cells. Both As4S4 and As2S3 can down-regulate the TF antigen , TF mRNA transcription and membrane PCA of NB4 and MR2 cells in vitro in a time-dependent manner. The role of As4S4 was stronger than that of As2S3. Both As4S4 and As2S3 had no effect on PML/RARa fusion gene transcription. CHX treatment completely suppressed the down-regulate effect of As4S4 on the TF mRNA expression. Conclusion: As4S4 and As2S3 may down regulate tissue factor expression and PCA of NB4 and MR2 cells. By down-regulating TF expression, As4S4 and As2S3 might be used to improve the DIC-related hemorrhage in APL patients. Elevated TF antigen level of NB4 and MR2 cells may be related to the fusion gene PML/RARa. The modulation of the TF mRNA expression in NB4 and MR2 cells by As4S4 and As2S3 might be indirect and might not involve PML/RARa fusion gene.展开更多
文摘Objective： To investigate the As4S4 induced growth inhibition and apoptosis in HeLa cells and its possible relationship with cyclooxygenase-2 （COX-2）. Methods： HeLa cells were treated with various concentrations （7.5, 15, 30, 60 mg/L） of As4S4 at different times （12, 24, 36, 48, 60 h）. Cell growth was measured by MTT. Apoptosis was detected by double staining flow cytometry （FCM）. Levels of PGE2 were measured by radioimmunoassay. The expression of COX-2 protein was examined by Western blot analysis. Results： After treated with different concentrations of As4S4, the growth of HeLa cells was suppressed significantly in a dose-and time-dependent manner. The IC50 of 24 h was 30 mg/L （P〈0.01）. As4S4 induced apoptosis with apoptosis rates at 8.13%-62.36% by flow cytometry （FCM） in a dose-dependent manners. The release of PGE2 was reduced in HeLa cells with the values being （70.56±2.03）, （48.58±2.28）, （29.25±1.57） and （18.02±1.04） respectively, significantly different compared with control group （3.15±0.01） （P〈0.01）. As4S4 also inhibited the activity and expression of COX-2 in a dose dependent manner and down-regulated the expression of COX-2 protein greatly. Conclusion： As4S4 could inhibit the proliferation and increase apoptosis in human HeLa cells. These effects may depend on the inhibition of the expression of COX-2 and PGE2 by As4S4.
基金supported by the National Natural Science Foundation of China(No.81403069)the Research and Development Fund of Beijing University of Chinese Medicine(No.2020072120050)。
文摘Mineral medicine,especially those containing heavy metals,is one of the characteristics of traditional Chinese medicine.A famous mineral medicine,realgar,containing heavy metal arsenic with a chemical formula of As4S4,has the function of detoxification,killing bacteria and viruses,and eliminating dampness and phlegm.Different As4S4 isomers are likely to have different drug effects and pharmacological actions.Therefore,it is of great scientific significance to find more stable As4S4 isomers.In view of this,ab initio molecular orbital theory and density functional theory(DFT)have been used to study ten isomers of As4S4 at the B3LYP/6-31 G*,B3LYP/6-311+G*,B3LYP/6-311+G(3 df,2 p)and MP2/(6-311+G*,LanL2 MB)levels of theory.In addition to the two isomers having been studied previously,eight new isomers were investigated in the present paper.All the ten As4S4 isomers were proved to be true local minima on their potential energy surfaces.The calculated NICS values and molecular orbital analyses showed that,the D2d symmetric As4S4,isomer 1,may be s-aromatic.The study proves that ten As4S4 isomers are stable thermodynamically,and are highly desirable for the future theoretical study of realgar.
基金Supported by Chinese Medicine Foundation of Shaanxi Province(No.1999002)
文摘Objective: To investigate the effect of arsenic sulfide (tetra-arsenic tetra-sulfide As4S4; diarsenic trisulfide As2S3) on tissue factor (TF) expression and procoagulant activity (PCA) of acute promyelocytic leukemia( APL) cell lines ( NB4 and MR2) and the basic mechanism of their role. Methods: NB4 and MR2 cells were respectively treated with As4S4 , As2S3, As4S4 and Cyclohexamide( CHX). PCA of the cells was detected using one-stage clotting assay. TF antigen was detected by ELISA. TF and PML/RARa fusion gene mRNA by semi-quantitive RT-PCR. The PCA and TF antigen of HL-60 and K562 cells were also examined. Results: The PCA and TF antigen level in NB4 and MR2 cells were significantly higher than that in HL-60 and K562 cells. Both As4S4 and As2S3 can down-regulate the TF antigen , TF mRNA transcription and membrane PCA of NB4 and MR2 cells in vitro in a time-dependent manner. The role of As4S4 was stronger than that of As2S3. Both As4S4 and As2S3 had no effect on PML/RARa fusion gene transcription. CHX treatment completely suppressed the down-regulate effect of As4S4 on the TF mRNA expression. Conclusion: As4S4 and As2S3 may down regulate tissue factor expression and PCA of NB4 and MR2 cells. By down-regulating TF expression, As4S4 and As2S3 might be used to improve the DIC-related hemorrhage in APL patients. Elevated TF antigen level of NB4 and MR2 cells may be related to the fusion gene PML/RARa. The modulation of the TF mRNA expression in NB4 and MR2 cells by As4S4 and As2S3 might be indirect and might not involve PML/RARa fusion gene.
