• The Journal of Korean Medicine
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• v.25 no.2
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• pp.33-40
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• 2004

Objective : This study investigated the apoptotic effect and its mechanism of Radix Aconiti (RA) extract and aconitine, which is a major constituent of RA, in HepG2 human hepatoma cells. Methods : We used MTT and DNA fragmentation assay to investigate cell viability and apoptotic effect on RA extract-treated HepG2 cells. In addition, to clarify the mechanism of RA extract-induced apoptosis, we applied caspase-3 enzyme activity assay and Western blotting method on poly-(ADP-ribose) polymerase (PARP) protein expression. Results : Treatment with RA extract resulted in the decrease of cell viability, and this effect was caused from apoptosis as confirmed by discontinuous fragmentation of DNA in HepG2 cells, but aconitine did not. Also, RA extract-treated HepG2 cells induced the activation of caspase-3 enzyme activity in time- and dose-dependent manners, which was accompanied by the cleavage of 116 kD PARP to 85 kD product. Conclusions : These results suggest that the apoptotic effects of RA extract on HepG2 cells could not be explained by aconitine. Additionally, RA extract induced apoptosis in hepatoma cells through caspase-3 activation and subsequent PARP cleavage.

• Biomedical Science Letters
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• v.12 no.4
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• pp.349-354
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• 2006

Induction of apoptosis allows the organism to get rid of abnormal cells and also of tumor cells. Understanding the mechanism involved in Ultraviolet radiation (UV) induced apoptosis may improve its therapeutic efficacy. In this study, we present expression of poly (ADP-ribose) polymerase (PARP) during apoptosis induced by UV in HeLa $S_3$ cells. Four different assays were performed in this study: morphological assessment of apoptotic cells and cell viability, DNA fragmentation analysis by agarose gel electrophoresis, quantitative assay of fragmented DNA, and expression of PARP by the western blot analysis. The percentages of apoptotic HeLa $S_3$ cells irradiated with $75J/m^2$ UV was increased continuously from 3 hrs incubation. DNA ladder pattern was appeared at 6 hrs. The amount of nucleosomal DNA fragments in cells treated UV increased from 3 to 12 hrs incubation and gradually decreased. The cleavage of PARP in HeLa $S_3$ cells irradiated with UV was induced, and the cleavage of PARP was more delayed in the cells pretreated with $5J/m^2$ UV and subsequently irradiated with $75J/m^2$ UV. than that in the cells only irradiated with $75J/m^2$ UV. Thus these data suggest that the cleavage of PARP relates with DNA fragmentation associated with apoptosis.

• Journal of Physiology & Pathology in Korean Medicine
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• v.24 no.3
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• pp.385-389
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• 2010

Butein(3,4,2',4-tetrahydroxychalcone) has been reported anticancer effects in several cancer type, which is prostate, bladder cancer but breast cancer is not. This study was to investigate the antiproliferative effects by butein(3,4,2',4-tetrahydroxychalcone) in MCF-7 human breast carcinoma cells. We invastigated the effects of dose-dependently cell growth inhibition by butein, which could be proved by WST-1 assay. Also, flow cytometry analysis was butein increase percentage of subG1 phase. As well as, butein induces apoptosis through the expression of caspase-8,-3 and poly(ADP-ribose) polymerase(PARP) activation but not in DMSO treated cells. Taken together, this results suggest that butein induced MCF-7 apoptosis through extrinsic pathway and thus may have potential tumor suppressor in breast cancer.

• YAKHAK HOEJI
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• v.47 no.5
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• pp.319-324
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• 2003

This study examined the inhibitory effect of extracts of Trichosanthes kirilpwii sorted according to the parts on the growth of HL-60 cells. The growth of HL-60 leukemia cells was markedly inhibited by the treatment of the 80％ methanol extract of roots (10 $\mu\textrm{g}$/mι), stems (50$\mu\textrm{g}$/mι), pips (10$\mu\textrm{g}$/mι), and gourds (100 $\mu\textrm{g}$/mι), or the ethylacetate fraction of leaves (100 $\mu\textrm{g}$/mι). when the HL-60 cells were treated with the extracts of T. kirilpwii sorted according to the parts, DNA fragmentation and sub-G1 hypodiploid cells were observed. Moreover, T. kirilpwii extracts increased the level of the expression of the active form of caspase-3 and the activation of caspase-3 was demonstrated by the cleavage of poly(ADP-ribose) polymerase, a vital substrate of effector caspase. The results suggest that the inhibitory effect of extracts of T. kirilpwii sorted according to the parts on the growth of HL-60 cells seems to arise from the induction of apoptosis.

• Tuberculosis and Respiratory Diseases
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• v.60 no.4
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• pp.451-463
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• 2006

Background : Reactive oxygen species (ROS) take center stage as executers in ventilator-induced lung injury (VILI). The protein with DNA-damage scanning activity, poly (ADP-ribose) polymerase-1 (PARP1), signals DNA rupture and participates in base-excision repair. Paradoxically,overactivation of PARP1 in response to massive genotoxic injury such as ROS can induce cell death through ${\beta}$ -nicotinamide adenine dinucleotide ($NAD^+$) depletion, resulting in inflammation. The purpose of this study is to investigate the role of PARP1 and the effect of its inhibitor in VILI. Methods : Forty-eight male C57BL/6 mice were divided into sham, lung protective ventilation(LPV), VILI, and PARP1 inhibitor (PJ34)+VILI (PJ34+VILI) groups. Mechanical ventilator setting for the LPV group was $PIP\;15cmH_2O$ + $PEEP\;3cmH_2O$ + RR 90/min + 2 hours. The VILI and PJ34+VILI groups were ventilated on a setting of $PIP\;40cmH_2O$ + $PEEP\;0cmH_2O$ + RR 90/min + 2 hours. As a PARP1 inhibitor for the PJ34+VILI group, 20 mg/Kg of PJ34 was treated intraperitoneally 2 hours before mechanical ventilation. Wet-to-dry weight ratio and acute lung injury (ALI) score were measured to determine the degree of VILI. PARP1 activity was evaluated by using an immunohistochemical method utilizing biotinylated NAD. Myeloperoxidase (MPO) activity and the concentration of inflammatory cytokines such as tumor necrosis factor $(TNF)-{\alpha}$, interleukin $(IL)-1{\beta}$, and IL-6 were measured in bronchoalveolar lavage fluid (BALF). Results : In the PJ34+VILI group, PJ34 pretreatment significantly reduced the degree of lung injury, compared with the VILI group (p<0.05). The number of cells expressing PARP1 activity was significantly increased in the VILI group, but significantly decreased in the PJ34+VILI group (p=0.001). In BALF, MPO activity, $TNF-{\alpha}$, $IL-1{\beta}$, and IL-6 were also significantly lower in the PJ34+VILI group (all, p<0.05). Conclusion : PARP1 overactivation plays a major role in the mechanism of VILI. PARP1 inhibitor prevents VILI, and decreases MPO activity and inflammatory cytokines.

• The Korean Journal of Physiology and Pharmacology
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• v.7 no.6
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• pp.325-331
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• 2003

3-Nitropropionic acid (3-NP) inhibits electron transport in mitochondria, leading to a metabolic failure. In order to elucidate the mechanism underlying this toxicity, we examined a few biochemical changes possibly involved in the process, such as metabolic inhibition, generation of reactive oxygen species (ROS), DNA strand breakage, and activation of Poly(ADP-ribose) polymerase (PARP). Exposure of SK-N-BE(2)C neuroblastoma cells to 3-NP for 48 h caused actual cell death, while inhibition of mitochondrial function was readily observed when exposed for 24 h to low concentrations (0.2${\sim}$2 mM) of 3-NP. The earliest biochemical change detected with low concentration of 3-NP was an accumulation of ROS (4 h after 3-NP exposure) followed by degradation of DNA. PARP activation by damaged DNA was also detectable, but at a later time. The accumulation of ROS and DNA strand breakage were suppressed by the addition of glutathione or N-acetyl-L-cysteine (NAC), which also partially restored mitochondrial function and cell viability. In addition, inhibition of PARP also reduced the 3-NP-induced DNA strand breakage and cytotoxicity. These results suggest that oxidative stress and activation of PARP are the major factors in 3-NP-induced cytotoxicity, and that the inhibition of these factors may be useful in protecting neuroblastoma cells from 3-NP-induced toxicity.

• Asian Pacific Journal of Cancer Prevention
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• v.17 no.8
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• pp.4107-4111
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• 2016

Background: Oral cancers account for approximately 2% of all cancers diagnosed each year; however, the vast majority (80%) of the affected individuals are smokers whose risk of developing a lesion is five to nine times greater than that of non-smokers. Tobacco smoke contains numerous carcinogens that cause DNA damage, including oxidative lesions that are removed effectively by the base-excision repair (BER) pathway, in which poly (ADP-ribose) polymerase 1 (PARP-1), plays key roles. Genetic variations in the genes encoding DNA repair enzymes may alter their functions. Several studies reported mixed effects on the association between PARP-1 variants and the risk of cancer development. Till now no reported studies have investigated the association between PARP-1 variants and oral squamous cell carcinoma (OSCC) risk in an Indian population. Materials and Methods: In the present case control study 100 OSCC patients and 100 matched controls were genotyped using PARP1 single nucleotide peptides (SNP's) rs1136410 and rs3219090 using TaqMan assays. Results: The results indicated significantly higher risk with PARP1 rs1136410 minor allele "C" (OR=1.909; p=0.02942; CI, 1.060-3.439). SNP rs1136410 also showed significantly increased risk in patients with smoking habit at C/C genotype and at minor allele C. Conclusions: The PAPR-1 Ala762Val polymorphism may play a role in progression of OSCC. Larger studies with a greater number of samples are needed to verify these findings.

• Journal of Life Science
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• v.27 no.2
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• pp.164-171
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• 2017

Our previous study suggested that S-allylcysteine (SAC) inhibits the proliferation of the human cervical cancer cell line, HeLa, at least in part through the induction of apoptosis and cell cycle arrest. To further analyze the specific molecular mechanism(s) by which SAC mediates its antiproliferative effects, this study examined the role of SAC in regulating the protein expression of initiator caspase (caspase-9), effector caspases (caspase-3 and caspase-7), and poly-ADP-ribose polymerase (PARP) in HeLa. Western blot analysis showed that when cells were treated with 50 mM SAC for 48 hr, the expression of procaspase-3, -7, and -9 and PARP was reduced by 94%, 38%, 95%, and 64%, respectively, as compared to the untreated control. In contrast, the expression of caspase-3, -7, and -9 and cleaved-PARP was markedly increased by SAC treatment. The SAC-mediated changes in the expression of these proteins were correlated with the concomitant inhibition of cellular proliferation by SAC. The cell proliferation assay showed that HeLa treatment with more than 20 mM SAC for 6-48 hr resulted in both concentration- and time-dependent inhibition of cellular proliferation. These results indicate that the SAC-induced antiproliferative effect in HeLa may be mediated at least in part through the activation of caspase-9, followed by the activation of caspase-3 and caspase-7 as well as the inactivation of PARP, thus leading to cellular apoptosis.

• The Korean Journal of Physiology and Pharmacology
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• v.26 no.4
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• pp.239-253
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• 2022

Epithelial-mesenchymal transition (EMT) is known to be involved in airway remodeling and fibrosis of bronchial asthma. However, the molecular mechanisms leading to EMT have yet to be fully clarified. The current study was designed to reveal the potential mechanism of microRNA-21 (miR-21) and poly (ADP-ribose) polymerase-1 (PARP-1) affecting EMT through the PI3K/AKT signaling pathway. Human bronchial epithelial cells (16HBE cells) were transfected with miR-21 mimics/inhibitors and PARP-1 plasmid/small interfering RNA (siRNA). A dual luciferase reporter assay and biotin-labeled RNA pull-down experiments were conducted to verify the targeting relationship between miR-21 mimics and PARP-1. The migration ability of 16HBE cells was evaluated by Transwell assay. Quantitative real-time polymerase chain reaction and Western blotting experiments were applied to determine the expression of Snail, ZEB1, E-cadherin, N-cadherin, Vimentin, and PARP-1. The effects of the PI3K inhibitor LY294002 on the migration of 16HBE cells and EMT were investigated. Overexpression of miR-21 mimics induced migration and EMT of 16HBE cells, which was significantly inhibited by overexpression of PARP-1. Our findings showed that PARP-1 was a direct target of miR-21, and that miR-21 targeted PARP-1 to promote migration and EMT of 16HBE cells through the PI3K/AKT signaling pathway. Using LY294002 to block PI3K/AKT signaling pathway resulted in a significant reduction in the migration and EMT of 16HBE cells. These results suggest that miR-21 promotes EMT and migration of HBE cells by targeting PARP-1. Additionally, the PI3K/AKT signaling pathway might be involved in this mechanism, which could indicate its usefulness as a therapeutic target for asthma.

• Journal of Applied Biological Chemistry
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• v.54 no.2
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• pp.143-146
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• 2011

Novel pyrazoline (4) was synthesized from chalcone (3) which was prepared from 2'-hydroxy-l'acetonaphthone (1) and 4-methoxy benzaldehyde (2). Pyrazoline (4) forms resonance assisted hydrogen bond between naphthol hydroxyl group and imine nitrogen in a pyrazoline ring. Pyrazoline (4) shows Poly ADP-ribose Polymerase (PARP) cleavage ability as a proof of apoptosis in cancer cell, which reveals its anti-cancer property.