The effect and mechanism of DADS inhibiting the proliferation and inducing apoptosis of gastro-esophageal adenocarcinoma cell line OE19 / 西安交通大学学报(医学版)

Objective To investigate the effect and mechanism of DADS in inhibiting the proliferation and inducing apoptosis of gastro-esophageal cancer cells in vitro.Methods The gastro-esophageal adenocarcinoma cells OE1 9 were treated by DADS of different concentrations in vitro.Morphologic changes were observed by the microscope and MTT assay was performed to test the growth-inhibitory effect of DADS on OE1 9 cells.Apoptosis rate of OE1 9 treated with different concentrations of DADS was measured by flow cytometry.Real-time PCR was used to detect DADS-induced effects on mRNA expressions of Caspase-3 ,Caspase-9 ,Bcl-2 ,Bax and NF-κB in OE1 9 cells.Results DADS inhibited the proliferation of OE19 cells in a dose-dependent manner.The apoptosis rate of OE19 cells was 14.0%,25.4% and 19.0% and 27.2%,respectively,when treated with 40 and 80μg/mL DADS for 24 h and 48 h.Real-time PCR assay showed that DADS could enhance mRNA expression levels of Caspase-3 and Caspase-9 and significantly decrease the mRNA expression levels of NF-κB and Bcl-2 and the ratio of Bcl-2/Bax. Conclusion DADS can significantly inhibit the proliferation and induce the apoptosis of gastro-esophageal adenocarcinoma cells via mitochondria-dependent pathways,which may be related to NF-κB and Bcl-2 families.

Diallyl disulfide induces apoptosis in human leukemia K562 cells through activation of NADPH oxidase / 中国药理学通报

Aim Toresearchthemolecularmecha-nisms of DADS-induced apoptosis in human leukemia K562cells.Methods Cellviabilitywasmeasuredby MTT. Levels of DADS-induced ROS were measured by 2ˊ, 7ˊ-dichlorofluorescein diacetate ( DCFH-DA) fluo-rescence. DADS-induced mRNA levels of components of the NADPH oxidase were detected by Real-time PCR. The combination of protein Rac2 and p67phox was measured by immunoprecipitation assays. Flow cy-tometry methods were used to determine the percentage of apoptosis cells. DADS-induced Rac2 levels were measuredbyWesternblot.Results TheDADS-trea-ted K562 cells showed a dose-and time-dependent de-crease in cell viability and proliferation. There was sig-nificant up-regulation of the mRNA level of components of the NADPH oxidase complex in K562 cells after treatment with 6 mg·L-1 DADS for 6 h. Western blot results revealed that, compared with the control group, there was a significant up-regulation of Rac2 protein in K562 cells treated with 5. 0 and 10. 0 mg·L-1 DADS for 24h. And Rac2 combined with p67phox in DADS-induced apoptosis in K562 cells. PMA markedly in-creased the percentage of apoptotic cells, and DPI re-duced the percentage of apoptotic cells in DADS-in-duced K562 cells. Levels of DADS-induced ROS, also showed enhancement when exposed in PMA, but there was no DADS-induced ROS production evident when exposed in DPI in DADS induced K562 cells. Conclu-sions TheseresultsindicatethatNADPHoxidaseis the main source of DADS-induced ROS production. Diallyl disulfide induces apoptosis in human leukemia K562 cells through activation of NADPH oxidase.

Role of ERK/AP-1 pathway in the differentiation induced by DADS on human gastric caner cell / 中国药理学通报

AIM To investigate the role of ERK/AP-1 pathway in the differentiation induced by diallyl disulfide (DADS) on human gastric cancer MGC803 cell. METHODS Immunocytochemical staining and morphometric quantitative analysis detected the expression of c-fos and c-jun;Western Blot which measured the activation of ERK was analyzed to elucidate the possible mechanism of DADS-induced human gastric cancer cell differentiation. RESULTS Immunocytochemical staining and morphometric quantitative analysis indicated that expression of c-fos and c-jun reduced(P

Effects of allyl sulfur compounds and garlic extract on the expression of Bcl-2, Bax, and p53 in non small cell lung cancer cell lines

Allyl sulfur compounds play a major role in the chemoprevention against carcinogenesis. The present study compared the antiproliferative effects of diallyl sulfide (DAS), diallyl disulfide (DADS) and garlic extract on p53-wild type H460 and p53-null type H1299 non small cell lung cancer cells (NSCLC). The DAS and DADS treatment of both H460 and H1299 cells resulted in the highest numbers of cells in apoptotic state as measured by acridine orange staining, however, garlic extract treatment did not induce any significant apoptotic cells by MTT assay. DADS was found to be more effective in inducing apoptosis on NSCLC. The level of p53 protein in H460 cell was increased following DADS treatment. DAS and garlic extract treatment of H460 cells induced a rise in the level of Bax and a fall of Bcl-2 level. These results demonstrate that DAS, DADS and garlic extract are effective in reduction of anti-proliferative gene in NSCLC and suggest that modulation of apoptosis-associated cellular proteins by DAS, DADS and garlic extract may be the mechanism for apoptosis which merit further investigation as potential chemoprevention agents.

Role of histone acetylation in the differentiation of human gastric cancer cell induced by DADS in vivo / 中国药理学通报

Aim To observe effect of DADS on the differentiation of gastric adenocarcinoma cells and acetylation of human gastric cancer tumor cell transplantable histone.Methods Human gastric adenocarcinoma heterotransplantable tumor model was constructed through subcutaneously injecting MGC803 cells to nude mice.Morphologic changes of xenograft tumor cells were observed with optical microscope,the influence of DADS on xenograft tumor cells generationcycle distribution,the expression of p21~(WAF1) protein,histone H3 and and H4 acetylion were analyzed with flow cytometry and Western blot.Results There was obvious growth inhibitory effect of xenograft tumor while abdominal injection dose were 100 and 200 mg?kg~(-1)DADS;cells density and heteromorphism decreased after treated with DADS.Flow cytometry analysis revealed that treating xenograft tumor cells with increasing quantities of DADS increased the percentage of cells in the G_2/M phase.The proportion of xenograft tumor cells in the G_2/M phase after treatment with 100 mg?kg~(-1) and 200 mg?kg~(-1) DADS was 2.22 and 3.37 times of that in NS group.Western blot analysis showed H3 acetylion increase along with G_2/M arrest of xenograft tumor cells by DADS.DADS didn′t influence the expression level of H4 acetylion;the expression of p21~(WAF1) protein in xenograft tumor increased along with the increases in the concentration of DADS.Conclusion DADS cansignificantly inhibit the growth of human gastric carcinoma xenograft in BALB/C nucle mice and induce cell differentiation,which might be related with up-regulation of histone a cetylization and p21~(WAF1) protein level.

Induction of differentiation by diallyl disulfide through inhibition of JAK1/STAT3 in human leukemia HL-60 cells / 中国药理学通报

Aim To investigate JAKs/STATs signal transduction change in HL-60 cells differentiation induced by diallyl disulfide(DADS)and molecular mechanism regulating the differentiation.Methods After incubation of HL-60 cells with DADS or AG490(50 ?mol?L -1),the cell differentiation indexes were observed by cytomorphology, NBT reduction ability assay,cell myeloid differentiation antigen CD11b by flow cytometry. Kinase activity of JAKs/STATs was tested by western-blotting and expressions of nucleus transcription genes stats,c-myc,c-fos,c-jun were detected by immumocyte chemistry method.Results Cell differentiation index changes indicated that HL-60 cells were induced differentiation toward granulocytic lineage by DADS, Western blot test demonstrated that constitive phosphorylation of Jak1,stat3 kinase was suppressed. Stat3,c-myc gene expression decreased and c-fos, c-jun gene expression increased in HL-60 cells treated with DADS through immunocyte chemistry.Conclusion Inhibition of phosphative Jak1, Stat3 was involved in HL-60 cells differentiation induced by DADS, its molecular mechanism might be related to modulation of gene expression associated proliferation and differentiation,and inhibition of DNA systhesis, induction differentiation.