Objective To evaluate the chemical composition and effects of Artemisia vulgaris(AV)hydroalcoholic extract(HEAV)on breast cancer cells(MCF-7 and SKBR-3),chronic myeloid leukemia(K562)and NIH/3T3 fibroblasts.Methods Ph...Objective To evaluate the chemical composition and effects of Artemisia vulgaris(AV)hydroalcoholic extract(HEAV)on breast cancer cells(MCF-7 and SKBR-3),chronic myeloid leukemia(K562)and NIH/3T3 fibroblasts.Methods Phytochemical analysis of HEAV was done by high-performance liquid chromatography-mass(HPLC)spectrometry.Viability and cell death studies were performed using trypan blue and Annexin/FITC-7AAD,respectively.Ferrostatin-1(Fer-1)and necrostatin-1(Nec-1)were used to assess the mode of HEAV-induced cell death and acetoxymethylester(BAPTA-AM)was used to verify the involvement of cytosolic calcium in this event.Cytosolic calcium measurements were made using Fura-2-AM.Results HEAV decreased the viability of MCF-7,SKBR-3 and K562 cells(P<0.05).The viability of HEAV-treated K562 cells was reduced compared to HEAV-exposed fibroblasts(P<0.05).Treatment of K562 cells with HEAV induced cell death primarily by late apoptosis and necrosis in assays using annexin V-FITC/7-AAD(P<0.05).The use of Nec-1 and Fer-1 increased the viability of K562 cells treated with HEAV relative to cells exposed to HEAV alone(P<0.01).HEAV-induced Ca^(2+)release mainly from lysosomes in K562 cells(P<0.01).Furthermore,BAPTA-AM,an intracellular Ca^(2+)chelator,decreased the number of non-viable cells treated with HEAV(P<0.05).Conclusions HEAV is cytotoxic and activates several modalities of cell death,which are partially dependent on lysosomal release of Ca^(2+).These effects may be related to artemisinin and caffeoylquinic acids,the main compounds identified in HEAV.展开更多
Pentachlorophenol（PCP） is a widespread,persistent environmental contaminant,and it is enzymatically activated to form a reactive metabolite,tetrachloro-l,4-benzoquinone（TCBQ）.To our knowledge,there is no informati...Pentachlorophenol（PCP） is a widespread,persistent environmental contaminant,and it is enzymatically activated to form a reactive metabolite,tetrachloro-l,4-benzoquinone（TCBQ）.To our knowledge,there is no information about TCBQ toxicity on embryonic stem cells.Here,we demonstrated that TCBQ induced significantly apoptosis of mouse embryonic stem cells in a concentration-dependent manner.We also showed that TCBQ elevated genomic5-hydroxymethylcytosine（5hmC） by affecting ten-eleven translocation（Tet） dioxygenases in mouse embryonic stem cells.We further investigated whether Tet dioxygenases were implicated in TCBQ-induced apoptosis.By depleting all three dioxygenases（Tet1-3）,we found that Tet dioxygenases slightly inhibited both early and late apoptosis induced by TCBQ at a low concentration（30 μmol/L）.Meanwhile,treated by TCBQ at higher concentrations（40and 50 μmol/L）,the total percentage of apoptotic cells was not affected by Tet dioxygenases.However,Tet dioxygenases tended to arrest mouse ES cells to be at early apoptotic stage and to reduce the cells to enter later apoptotic stage.These results indicate that Tet dioxygenases play a role in shaping TCBQ-induced apoptosis in mouse embryonic stem cells.Our study provides new insights into the toxicology of PCP and its reactive metabolite TCBQ.展开更多
基金Supported by the São Paulo Research Foundation[FAPESP,No.2020/14406(CB)]Brazilian National Council for Scientific and Technological Development(CNPq,No.PQ-302148/2019-1)Coordination of Superior Level Staff Improvement(CAPES)。
文摘Objective To evaluate the chemical composition and effects of Artemisia vulgaris(AV)hydroalcoholic extract(HEAV)on breast cancer cells(MCF-7 and SKBR-3),chronic myeloid leukemia(K562)and NIH/3T3 fibroblasts.Methods Phytochemical analysis of HEAV was done by high-performance liquid chromatography-mass(HPLC)spectrometry.Viability and cell death studies were performed using trypan blue and Annexin/FITC-7AAD,respectively.Ferrostatin-1(Fer-1)and necrostatin-1(Nec-1)were used to assess the mode of HEAV-induced cell death and acetoxymethylester(BAPTA-AM)was used to verify the involvement of cytosolic calcium in this event.Cytosolic calcium measurements were made using Fura-2-AM.Results HEAV decreased the viability of MCF-7,SKBR-3 and K562 cells(P<0.05).The viability of HEAV-treated K562 cells was reduced compared to HEAV-exposed fibroblasts(P<0.05).Treatment of K562 cells with HEAV induced cell death primarily by late apoptosis and necrosis in assays using annexin V-FITC/7-AAD(P<0.05).The use of Nec-1 and Fer-1 increased the viability of K562 cells treated with HEAV relative to cells exposed to HEAV alone(P<0.01).HEAV-induced Ca^(2+)release mainly from lysosomes in K562 cells(P<0.01).Furthermore,BAPTA-AM,an intracellular Ca^(2+)chelator,decreased the number of non-viable cells treated with HEAV(P<0.05).Conclusions HEAV is cytotoxic and activates several modalities of cell death,which are partially dependent on lysosomal release of Ca^(2+).These effects may be related to artemisinin and caffeoylquinic acids,the main compounds identified in HEAV.
基金supported by the National Natural Science Foundation of China(Nos.21327006,21435008,21321004)the Strategic Priority Research Program of the Chinese Academy of Sciences(No.XDB14030200)
文摘Pentachlorophenol（PCP） is a widespread,persistent environmental contaminant,and it is enzymatically activated to form a reactive metabolite,tetrachloro-l,4-benzoquinone（TCBQ）.To our knowledge,there is no information about TCBQ toxicity on embryonic stem cells.Here,we demonstrated that TCBQ induced significantly apoptosis of mouse embryonic stem cells in a concentration-dependent manner.We also showed that TCBQ elevated genomic5-hydroxymethylcytosine（5hmC） by affecting ten-eleven translocation（Tet） dioxygenases in mouse embryonic stem cells.We further investigated whether Tet dioxygenases were implicated in TCBQ-induced apoptosis.By depleting all three dioxygenases（Tet1-3）,we found that Tet dioxygenases slightly inhibited both early and late apoptosis induced by TCBQ at a low concentration（30 μmol/L）.Meanwhile,treated by TCBQ at higher concentrations（40and 50 μmol/L）,the total percentage of apoptotic cells was not affected by Tet dioxygenases.However,Tet dioxygenases tended to arrest mouse ES cells to be at early apoptotic stage and to reduce the cells to enter later apoptotic stage.These results indicate that Tet dioxygenases play a role in shaping TCBQ-induced apoptosis in mouse embryonic stem cells.Our study provides new insights into the toxicology of PCP and its reactive metabolite TCBQ.
