1.4-(Tert-butyl)-2,6-bis(1-phenylethyl)phenol induces pro-apoptotic activity.
Jun Ho KIM ; Yunmi LEE ; Mi Yeon KIM ; Jae Youl CHO
The Korean Journal of Physiology and Pharmacology 2016;20(3):253-259
Previously, we found that KTH-13 isolated from the butanol fraction of Cordyceps bassiana (Cb-BF) displayed anti-cancer activity. To improve its antiproliferative activity and production yield, we employed a total synthetic approach and derivatized KTH-13 to obtain chemical analogs. In this study, one KTH-13 derivative, 4-(tert-butyl)-2,6-bis(1-phenylethyl)phenol (KTH-13-t-Bu), was selected to test its anti-cancer activity. KTH-13-t-Bu diminished the proliferation of C6 glioma, MDA-MB-231, LoVo, and HCT-15 cells. KTH-13-t-Bu induced morphological changes in C6 glioma cells in a dose-dependent manner. KTH-13-t-Bu also increased the level of early apoptotic cells stained with annexin V-FITC. Furthermore, KTH-13-t-Bu increased the levels of cleaved caspase-3 and -9. In contrast, KTH-13-t-Bu upregulated the levels of pro- and cleaved forms of caspase-3, -8, and -9 and Bcl-2. Phospho-STAT3, phospho-Src, and phospho-AKT levels were also diminished by KTH13-t-Bu treatment. Therefore, these results strongly suggest that KTH-13-t-Bu can be considered a novel anti-cancer drug displaying pro-apoptotic activity.
Caspase 3
;
Cordyceps
;
Glioma
2.The Effect of Cysteamine on the Radiation-Induced Apoptosis.
Young Min CHOI ; Chang Gyo PARK ; Heung Lae CHO ; Hyung Sik LEE ; Won Joo HUR
The Journal of the Korean Society for Therapeutic Radiology and Oncology 2000;18(3):214-219
PURPOSE: To investigate the pathways of radiation induced apoptosis and the effect of cysteamine (beta-mercaptoethylamine), as a radioprotector, on it. MATERIALS AND METHODS: HL-60 cells were assigned to control, irradiated, and cysteamine (1 mM, 10mM) pretreated groups, Irradiation was given in a single fraction of 10 Gy (6 MV x-ray) and cysteamine was administered 1 hour before irradiation. The activities of caspase-8 were measured in control and irradiated group to evaluate its relation to the radiation induced apoptosis. To evaluate the role of cysteamine in radiation induced apoptosis, the number of viable cells, the expression and activity of caspase-3, and the expression of poly (ADP-ribose) polymerase (PARP) were measured and compared after irradiation the HL-60 cells with cysteamine pretreatment or not. RESULTS: The intracellular caspase-8 activity, known to be related to the death receptor induced apoptosis, was not affected by irradiation (p>0.05). The number of viable cells began to decrease from 6 hours after irradiation (p>0.05), but the number of viable cells in 1 mM cysteamine pretreated group was not decreased afger irradiation and was similar to those in the control group. In caspase-3 analyses, known as apoptosis executioner, its expression was not different but its activity was increased by irradiation (p>0.05). However, this increase of activity was suppressed by the pretreatment of 1mM crysteamine. The cleavage of PARP, thought to be resulted from caspase-3 activation, occurred after irradiation, which was attenuated by the pretreatment of 1mM cysteamine. CONCLUSION: these results show that radiation induced apoptotic process is somewhat different from death receptor induced one and the pretreatment of 1 mM cysteamine has a tendency to decrease the radiation-induced apoptosis in HL-60 cells.
Apoptosis*
;
Caspase 3
;
Caspase 8
;
Cysteamine*
;
HL-60 Cells
;
Humans
3.The Antiapoptotic Effect of Danggui Buxue Tang and Its Main Components on Hematopoietic Cells in Mice with Bone Marrow Suppression.
Hui-Min KONG ; Wei-Qing SU ; Hong YE ; Hua WANG ; Liang LI ; Hui CHEN ; Mo YANG
Journal of Experimental Hematology 2022;30(6):1679-1687
OBJECTIVE:
To explore the hematopoiesis protection effect of Danggui Buxue Tang (DBT) and its main components Angelica polysaccharide (APS) and Astragalus polysaccharide (ASPS) on myelosuppression mice, and the mechanism of anti-apoptosis of Meg-01 cells.
METHODS:
Mice were radiated with 4 Gy of 137Csγ ray to establish the model of radiation-induced myelosuppression. DBT, APS or ASPS (10 mg/kg) were injected into irradiated mice. Peripheral blood cell counts were performed on mice before radiation (day 0) and day 7, 14 and 21 after radiation. On the 21st day, poor plasma platelets were collected from mice to detect TPO concentration and then the mice were sacrificed. The femoral bone marrow cells were cultured for colony cell forming units (CFU). Meg-01 cells were cultured without FBS for 24 h to induce apoptosis, and then treated with DBT/APS/ASPS for 72 h. Flow cytometry (FCM) was used to detect early apoptosis (Annexin V), mitochondrial membrane potential (JC-1) and the expression of Caspase-3 to analyze the effect of DBT/APS/ASPS on cell apoptosis.
RESULTS:
DBT can stimulate the recovery of white blood cells (WBC), red blood cells (RBC) and platelets (PLT) of myelosuppression mice, especially for WBC and PLT (P<0.01, P<0.05). Compared with the control group, the number of BFU-E, CFU-MK and CFU-GM increased after adding DBT (BFU-E & CFU-GM: P<0.05; CFU-MK: P<0.01). The effect of DBT on blood TPO concentration in mice was not obvious (P=0.89). RBC, WBC and PLT were increased in APS group compared with control group (P<0.05). WBC increased after the treatment of ASPS (P<0.05). APS stimulated the formation of CFU-F, CFU-MK and CFU-GM (P<0.05). Only CFU-GM increased in ASPS group(P<0.05). Besides, DBT decreased the apoptosis of Meg-01 cells (P<0.05). The early apoptosis rate and total death rate in APS (100 μg/ml) group were lower than that of control group (P<0.01, P<0.05). The early apoptosis rate of ASPS (100 μg/ml) group was lower than that of control group (P<0.05). JC-1 and Caspase-3 showed that APS (100 μg/ml) significantly reduced apoptosis rate (P<0.01, P<0.05).
CONCLUSION
DBT has protective effect on hematopoietic system, especially WBC and PLT, and has anti-apoptotic effect on Meg-01. It was found that the above effects of DBT were mainly caused by APS, and its anti-apoptosis mechanism was carried out mainly through JC-1 and Caspase-3 pathways.
Mice
;
Animals
;
Caspase 3
;
Bone Marrow
;
Polysaccharides
4.Pro-Apoptotic Activity of 4-Isopropyl-2-(1-Phenylethyl) Aniline Isolated from Cordyceps bassiana.
Mi Seon KIM ; Yunmi LEE ; Gi Ho SUNG ; Ji Hye KIM ; Jae Gwang PARK ; Han Gyung KIM ; Kwang Soo BAEK ; Jae Han CHO ; Jaegu HAN ; Kang Hyo LEE ; Sungyoul HONG ; Jong Hoon KIM ; Jae Youl CHO
Biomolecules & Therapeutics 2015;23(4):367-373
Cordyceps species including Cordyceps bassiana are a notable anti-cancer dietary supplement. Previously, we identified several compounds with anti-cancer activity from the butanol fraction (Cb-BF) of Cordyceps bassiana. To expand the structural value of Cb-BF-derived anti-cancer drugs, we employed various chemical moieties to produce a novel Cb-BF-derived chemical derivative, KTH-13-amine-monophenyl [4-isopropyl-2-(1-phenylethyl) aniline (KTH-13-AMP)], which we tested for anti-cancer activity. KTH-13-AMP suppressed the proliferation of MDA-MB-231, HeLa, and C6 glioma cells. KTH-13-AMP also dose-dependently induced morphological changes in C6 glioma cells and time-dependently increased the level of early apoptotic cells stained with annexin V-FITC. Furthermore, the levels of the active full-length forms of caspase-3 and caspase-9 were increased. In contrast, the levels of total forms of caspases-3, caspase-8, caspase-9, and Bcl-2 were decreased in KTH-13-AMP treated-cells. We also confirmed that the phosphorylation of STAT3, Src, and PI3K/p85, which is linked to cell survival, was diminished by treatment with KTH-13-AMP. Therefore, these results strongly suggest that this compound can be used to guide the development of an anti-cancer drug or serve as a lead compound in forming another strong anti-proliferative agent.
Apoptosis
;
Caspase 3
;
Caspase 8
;
Caspase 9
;
Cell Survival
;
Cordyceps*
;
Dietary Supplements
;
Glioma
;
Phosphorylation
5.Expression of IAP Genes During TPA-Induced Differentiation of HL-60 Cells.
Won Ki BAEK ; Taeg Kyu KWON ; Jong Wook PARK ; Min Ho SUH
Journal of Bacteriology and Virology 2003;33(1):113-118
HL-60 cells (human promyelocytic leukemia cells) differentiate into the monocyte/macrophage like cells that die spontaneously by apoptosis when treated with 12-O-tetradecanoylphorbol-13-acetate (TPA). It is known that inhibitors of apoptosis proteins (IAP) bind to and inhibit caspase 3, 7, 9 activity and the induction of apoptosis. In this study, we examined the expression of IAP genes during TPA induced differentiation of HL-60 cells. During the differentiation, HIAP-1, HIAP-2, and XIAP expressions were decreased in protein levels. The pan-caspase inhibitor z-VAD-fmk blocked the decrease of HIAP-1 and HIAP-2, which indicates HIAP-1 and HIAP-2 could be caspase substrates. These findings suggest that the decrease of IAP proteins is related to the induction of apoptosis that is associated with TPA- induced HL-60 cell differentiation.
Apoptosis
;
Caspase 3
;
HL-60 Cells*
;
Humans
;
Leukemia
6.Induction of Apoptosis by Vibrio vulnificus Cytolysin Through Activation of Caspase-3 in Endothelial Cells.
Byung Hyun PARK ; Kang Beom KWON ; Young Hoon LEE ; Jae Han JANG ; Jin Woo PARK
Journal of Bacteriology and Virology 2001;31(4):333-341
No abstract available.
Apoptosis*
;
Caspase 3*
;
Endothelial Cells*
;
Perforin*
;
Vibrio vulnificus*
;
Vibrio*
7.Ad-NK4 Enhances the Chemosensitivity of Human Multiple Myeloma RPMI 8226 Cells to Bortezomib.
Journal of Experimental Hematology 2016;24(4):1079-1085
OBJECTIVETo investigate if Ad-NK4 can enhance the chemosensitivity of human multiple myeloma RPMI 8226 cells to bortezomib(BOR).
METHODSThe cell-matrix adhesion test and PRMI 8226 cell-ECV 304 cell adhesion test were used to analyze the effect of Ad-NK4 on adhesion of RPMI 8226 cells; Western blot was used to detect the expression changes of adhesion and invasion-associated proteins MMP2, MMP3, MMP7 and VEGF; MTT assay was used to detect the effect of Ad-NK4 on proliferation of RPMI 8226 cells; the flow cytometry with PI staining was used to detect the effect of Ad-NK4 on cell apoptosis; the expression of cleaved caspase-3, BAX and BCL-2 was assayed by Western blot.
RESULTSThese 2 adhesion assays indicated that Ad-NK4 significantly inhibited the adhesion of human multiple myeloma RPMI 8226 cells. In addition, Erk and JAK/STAT pathway may be involved in the process. The expression level of MMP-2, MMP-3 and VEGF were decreased in Ad-NK4 group, compared with untreated or Ad-GFP group (P<0.05). However, the expression of MMP-7 protein in Ad-NK4 group was not significantly different from untreated or Ad-GFP group (P>0.05). The inhibitory rates of the proliferation in cells treatedly Ad-NK4 combined with BOR was significantly higher than that with BOR or Ad-NK4 alone. Similarly, Western blot indicated that the level of cleaved caspase-3 and BAX in cells treated with Ad-NK4 combined with BOR was significantly higher than BOR or Ad-NK4 alone, but BCL-2 protein expression was significantly lower. Meanwhile, the ratio of BAX/BCL-2 was increased.
CONCLUSIONAd-NK4 can enhance the chemosensitivity of human multiple myeloma RPMI 8226 cells to BOR,which is associated with increasing of both BAX/BCL-2 ratio and Caspase-3 activation.
Apoptosis ; Bortezomib ; Caspase 3 ; Cell Line, Tumor ; Humans ; Multiple Myeloma
8.The Hematopoietic Protective Effect of PDGF-BB on Radiation-Induced Myelosuppression Model Mice.
Yi LUO ; Hui-Min KONG ; Wei-Qing SU ; Wen-Fang YI ; Hui GE ; Hui CHEN ; Liang LI ; Mo YANG
Journal of Experimental Hematology 2022;30(6):1873-1880
OBJECTIVE:
To investigate the hematopoietic protective effect of platelet-derived growth factor (PDGF)-BB on radiation-induced myelosuppression model mice and effect of anti-apoptosis of megakaryocyte line Meg-01 cells, and its possible mechanism.
METHODS:
Mice were radiated with 4 Gy of 137Csγ ray to establish the model of myelosuppression. Mice were weighed and peripheral blood cell were counted before radiation (day 0) and day 7, 14 and 21 after radiation. On the 21 st day, the mice were killed. The sternal tissues of the mice were taken for morphological observation, and the femoral bone marrow cells were cultured for the assay of colony cell forming units (CFU). Meg-01 cells were cultured without FBS for 24 h to induce apoptosis, and then treated with PDGF-BB for 48 h. The effects of PDGF-BB on the proliferation were investigated by cell counting. Flow cytometry was used to detect early apoptosis (Annexin V), mitochondrial membrane potential (JC-1) and the expression of caspase-3.
RESULTS:
Peripheral blood cell counts of mice showed that PDGF-BB stimulated the recovery of white blood cells, red blood cells and platelets after radiation (P<0.05), especially for white blood cells. Morphological examination showed bone marrow hyperplasia in PDGF-BB group, the numbers of megakaryocytes and their progenitor cells were higher than those in the control group. PDGF-BB significantly stimulated the formation of CFU-MK, CFU-GM, BFU-E and CFU-F. PDGF-BB showed a strong proliferation effect in the concentration range of 5-50 ng/ml (P<0.001). PDGF-BB (50 ng/ml) significantly reduced the positive expression of Annexin V (P<0.01). The mitochondrial membrane potential in the control group was decreased when compared with PDGF-BB group, which indicated that the number of apoptotic cells was increased (P<0.01). Besides, the expression of caspase-3 in PDGF-BB group was significantly lower than that in control group (P<0.05).
CONCLUSION
PDGF-BB has a protective effect on the hematopoietic system of myelosuppression model mice, especially megakaryocytes and their progenitor cells. PDGF-BB has pro-proliferative and anti-apoptotic effects on Meg-01 cells, and the mechanism may be mediated through JC-1 and caspase-3 pathway.
Animals
;
Mice
;
Becaplermin
;
Caspase 3
;
Hematopoietic System
;
Apoptosis
9.Columbianadin Inhibits Cell Proliferation by Inducing Apoptosis and Necroptosis in HCT116 Colon Cancer Cells.
Ji In KANG ; Ji Young HONG ; Jae Sue CHOI ; Sang Kook LEE
Biomolecules & Therapeutics 2016;24(3):320-327
Columbianadin (CBN), a natural coumarin from Angelica decursiva (Umbelliferae), is known to have various biological activities including anti-inflammatory and anti-cancer effects. In this study, the anti-proliferative mechanism of actions mediated by CBN was investigated in HCT-116 human colon cancer cells. CBN effectively suppressed the growth of colon cancer cells. Low concentration (up to 25 μM) of CBN induced apoptosis, and high concentration (50 μM) of CBN induced necroptosis. The induction of apoptosis by CBN was correlated with the modulation of caspase-9, caspase-3, Bax, Bcl-2, Bim and Bid, and the induction of necroptosis was related with RIP-3, and caspase-8. In addition, CBN induced the accumulation of ROS and imbalance in the intracellular antioxidant enzymes such as SOD-1, SOD-2, catalase and GPx-1. These findings demonstrate that CBN has the potential to be a candidate in the development of anti-cancer agent derived from natural products.
Angelica
;
Apoptosis*
;
Biological Products
;
Caspase 3
;
Caspase 8
;
Caspase 9
;
Catalase
;
Cell Proliferation*
;
Colon*
;
Colonic Neoplasms*
;
Humans
;
Oxidative Stress
10.Anti-leukemic properties of deferasirox via apoptosis in murine leukemia cell lines.
Sol Rim JEON ; Jae Wook LEE ; Pil Sang JANG ; Nack Gyun CHUNG ; Bin CHO ; Dae Chul JEONG
Blood Research 2015;50(1):33-39
BACKGROUND: Although deferasirox (DFX) is reported to have anti-tumor effects, its anti-leukemic activity remains unclear. We evaluated the effect of DFX treatment on two murine lymphoid leukemia cell lines, and clarified the mechanisms underlying its potential anti-leukemic activity. METHODS: L1210 and A20 murine lymphoid leukemia cell lines were treated with DFX. Cell viability and apoptosis were evaluated by the 3-(4,5-dimethylthaizol-2-yl)-5-(3-carboxymethylphenyl)-2-(4-sulfophenyl)-2H-tetrazolium (MTS) assay and fluorescence-activated cell sorting (FACS) analysis, respectively. Immunoblotting was performed to detect the expression of key apoptotic proteins. RESULTS: In dose- and time-dependent manner, DFX decreased viability and increased apoptosis of murine leukemic cells. Fas expression was significantly higher in A20 cells than in L1210 cells at all DFX concentrations tested. Although both cell lines exhibited high caspase 3 and caspase 9 expression, a critical component of the intrinsic mitochondrial apoptotic pathway, expression was greater in L1210 cells. In contrast, caspase 8, a key factor in the extrinsic apoptotic pathway, showed greater expression in A20 cells. Cytochrome c expression was significantly higher in L1210 cells. In both cell lines, co-treatment with ferric chloride and DFX diminished the expression of these intracellular proteins, as compared to DFX treatment alone. CONCLUSION: Treatment with DFX increased caspase-dependent apoptosis in two murine lymphoid leukemia cell lines, with differing apoptotic mechanisms in each cell line.
Apoptosis*
;
Caspase 3
;
Caspase 8
;
Caspase 9
;
Cell Line*
;
Cell Survival
;
Cytochromes c
;
Flow Cytometry
;
Immunoblotting
;
Leukemia*
;
Leukemia, Lymphoid