• Title/Summary/Keyword: sub-G1 DNA

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Polysiponia morrowii Extract Inhibits Cancer Growth on CT-26 and Hela cells

  • Zhang, Chunying;Cha, Seon-Heui
    • Journal of Marine Bioscience and Biotechnology
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    • v.12 no.2
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    • pp.123-130
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    • 2020
  • Cancer is an unfavorable human disease, and the treatment commonly have side effects and can be ineffective. Since exploration and development of cancer treatment drugs is particularly demanding, this study aimed to investigate the anticancer activities of Polysiponia morrowii extract s (PME) on CT-26 and HeLa cells. The results showed that PME inhibited cell proliferation in a dose-dependent manner, with IC50 values of 41.04% in CT-26 and 48.51% in HeLa cell cultures. Moreover, cytological observation using Hoechst 33342 staining assay showed typical apoptotic morphology in both cancer cells, and production of sub-G1 DNA was induced by PME treatment in a dose-dependent manner, with 34.41% in CT-26 and 46.01% in HeLa cell cultures. These findings suggest that PME may have potential preventive effects or medicinal value in the treatment of colorectal and cervical cancers.

Resveratrol Induces Apoptosis in Primary Human Prostate Cancer Cells (Primary 인체 전립선 암세포에서 Resveratrol의 Apoptosis 유도 효과)

  • Kang, Hye-In;Kim, Jae-Yong;Cho, Hyun-Dong;Park, Kyung-Wuk;Kang, Jum-Soon;Seo, Kwon-Il
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.39 no.8
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    • pp.1119-1125
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    • 2010
  • To evaluate resveratrol as a prostate cancer preventive material, we investigated its anti-proliferative and apoptotic effects in RC-58T/h/SA#4 primary human prostate cancer cells. Resveratrol significantly decreased the number of viable RC-58T/h/SA#4 cells in a dose- and time-dependent manner. Resveratrol showed cytotoxicity against RC-58T/h/SA#4, LNCaP, PC-3 human prostate cancer cells with $IC_{50}$ values of 245, 320 and $340\;{\mu}M$, respectively. However the cytotoxic potential of resveratrol against normal RWPE-1 cells was lower ($IC_{50}=982\;{\mu}M$). Resveratrol induced cell death as evidenced by the increased formation of apoptotic bodies, nuclear condensation, sub-G1 phase, and DNA fragmentation. Resveratrol activated initiator caspases 8, and 9 as well as effector caspase 3 in a dose-dependent manner. Furthermore, the general caspase inhibitor z-VAD-fmk significantly inhibited resveratrol-induced apoptosis compared to cells without treatment. These results clearly indicate that resveratrol-induced apoptosis was dependent on caspase activation. Further, resveratrol modulated the down regulation of Bcl-2 (anti-apoptotic), and Bid. However, the level of Bax (pro-apoptotic) remained unchanged. These results suggest that resveratrol induced apoptosis in RC-58T/h/SA#4 cells via a mitochondrial-mediated caspase-dependent pathway, suggesting therapeutic potential against prostate cancer.

Apoptotic Cell Death of Human Leukemia U937 Cells by Essential Oil purified from Schisandrae Semen (오미자 종자 정유에 의한 인체백혈병 U937 세포의 apoptosis 유도)

  • Choi, Yung Hyun
    • Journal of Life Science
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    • v.25 no.2
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    • pp.249-255
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    • 2015
  • Schisandrae fructus [Schizandra chinensis (Turcz.) Baillon] is a medicinal herb widely used for treating various inflammatory and immune diseases in East Asian countries. The Schisandrae Semen essential oil (SSeo) from this plant has pharmacological activities, including antioxidant, antimicrobial, and antitumoral activities. Nevertheless, the biological activities and underlying molecular mechanisms of the potential anti-cancer effects of this oil remain unclear. In the present study, we investigated the potential inhibition of apoptosis signaling pathways by SSeo in human leukemia U937 cells and evaluated the underlying molecular mechanism. Exposure to SSeo resulted in a concentration-dependent growth inhibition due to apoptosis, which was verified by DNA fragmentation, the presence of apoptotic bodies, and an increase in the sub-G1 ratio. Induction of apoptotic cell death by SSeo was correlated with the down-regulation of members of the inhibitor of apoptosis protein (IAP) family (including X-linked inhibitor of apoptosis protein (XIAP), cIAP-1, and surviving) and anti-apoptotic Bcl-2, and with up-regulation of death receptor (DR) 4 and DR5, depending on dosage. SSeo treatment also induced Bid truncation, mitochondrial dysfunction, proteolytic activation of caspase-3, -8 and -9, and concomitant degradation of activated caspase-3 target proteins such as poly (ADP-ribose) polymerase. Taken together, these findings suggest that SSeo may be a potential chemotherapeutic agent for use in the control of human leukemia cells. Further studies are needed to identify its active compounds.

Induction of Apoptosis by Scolopendra subspinipes mutilans in Human Leukemia HL-60 Cells through Bcl-xL Regulation (왕지네(Scolopendra subspinipes mutilans)의 Bcl-xL 조절에 의한 HL-60 세포의 아폽토시스(Apoptosis) 유도)

  • Kim, Kil-Nam;Kim, Sang-Bum;Yoon, Weon-Jong;Yang, Kyoung-Sik;Park, Soo-Yeong
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.37 no.11
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    • pp.1408-1414
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    • 2008
  • The anticancer properties of Scolopendra subspinipes mutilans extract were investigated. The extract from S. subspinipes mutilans by 80% EtOH was fractionated with n-hexane, dichloromethan ($CH_2Cl_2$), ethylacetate (EtOAc), and butanol (BuOH) in order. The EtOAc fraction showed the highest inhibitory activity (about 80%) against human leukemia (HL-60) cell growth at $50\;{\mu}g/mL$. To explore the mechanism of cytotoxicity, we used several measures of apoptosis to determine whether these processes were involved in EtOAc fraction-induced HL-60 cell death. Our results showed EtOAc fraction induced cell shrinkage, cell membrane blebbing, apoptotic body, and DNA fragmentation. The EtOAc fraction gradually decreased the expression of anti-apoptotic Bcl-xL and led to the activation of caspase-3, -9 and cleavage of PARP. These findings suggest that S. subspinipes mutilans exhibits potential anticancer properties.

Molecular Biological Studies on Korean Garlic Viruses

  • Choi, Jin-Nam;Song, Jong-Tae;Shin, Chan-Seok;La, Yong-Joon;Lee, Jong-Seob;Choi, Yang-Do
    • Proceedings of the Korean Society of Plant Pathology Conference
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    • 1994.06a
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    • pp.86-102
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    • 1994
  • To understand the molecular structure and pathogenesis mechanism of Korean garlic viruses, we have isolate cDNA clones for garlic viruses. The partial nucleotide sequences of 24 cDNA clones were determined and that of six clones containing poly (A) tail were compared with those of other plant viruses. One of those clones, V9 has 81.8% similarity in nucleotide sequence and 93.0% in deduced amino acid sequence, respectively, to the coat protein gene for garlic mosaic virus (GMV). Northern blot analysis with the clone V9 demonstrated that the genome of GMV is 7.8 kb long and has poly (A) tail. The anti-coat protein antibody for GMV recognizes 35 kDa polypeptide which could be the coat protein of GMV from infected garlic leaf extract or virus preparation. Clone G7 has about 62% of deduced amino acid sequence identity with the members of potyvirus group. Northern blot analysis with the clone G7 demonstrated that the genome of the potyvirus I garlic is 9.0 kb long and has poly (A) tail. The third clone, S81, shows 42% amino acid identity to the potexvirus. The other clones are under the characterization. To test the possibility of producing garlic virus resistant plant, we have designed a hairpin type ribozyme to cleave V9 RNA at the middle of the coat protein gene. From the cleavage reactions in vitro with two different sizes of RNA substrates, V9SUB (144 nucleotides) and V9 RNA (1,361 nucleotides), the ribozyme can cleave V9 sequence effectively at the predicted site. To study the activity of the ribozyme in vivo, plant transformation is in progress. Further possibilities to produce garlic virus resistant plant will be discussed.

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Carrier frequency of SLC26A4 mutations causing inherited deafness in the Korean population

  • Kim, Hyogyeong;Lim, Hwan-Sub;Ryu, Jae-Song;Kim, Hyun-Chul;Lee, Sanghoo;Kim, Yun-Tae;Kim, Young-Jin;Lee, Kyoung-Ryul;Park, Hong-Joon;Han, Sung-Hee
    • Journal of Genetic Medicine
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    • v.11 no.2
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    • pp.63-68
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    • 2014
  • Purpose: The mutation of the SLC26A4 gene is the second most common cause of congenital hearing loss after GJB2 mutations. It has been identified as a major cause of autosomal recessive nonsyndromic hearing loss associated with enlarged vestibular aqueduct and Pendred syndrome. Although most studies of SLC26A4 mutations have dealt with hearing-impaired patients, there are a few reports on the frequency of these mutations in the general population. The purpose of this study was to evaluate the prevalence of SLC26A4 mutations that cause inherited deafness in the general Korean population. Materials and Methods: We obtained blood samples from 144 Korean individuals with normal hearing. The samples were subjected to polymerase chain reaction to amplify the entire coding region of the SLC26A4 gene, followed by direct DNA sequencing. Results: Sequencing analysis of this gene identified 5 different variants (c.147C>G, c.225G>C, c.1723A>G, c.2168A>G, and c.2283A>G). The pathogenic mutation c.2168A>G (p.H723R) was identified in 1.39% (2/144) of the subjects with normal hearing. Conclusion: These data provide information about carrier frequency for SLC26A4 mutation-associated hearing loss and have important implications for genetic diagnostic testing for inherited deafness in the Korean population.

Protective Effect of Korean Red Ginseng against Aflatoxin B1-Induced Hepatotoxicity in Rat

  • Kim, Yong-Seong;Kim, Yong-Hoon;Noh, Jung-Ran;Cho, Eun-Sang;Park, Jong-Ho;Son, Hwa-Young
    • Journal of Ginseng Research
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    • v.35 no.2
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    • pp.243-249
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    • 2011
  • Korean red ginseng (KRG), the steamed root of Panax ginseng Meyer, has a variety of biological properties, including anti-inflammatory, antioxidant and anticancer effects. Aflatoxin $B_1$ ($AFB_1$) produced by the Aspergillus spp. causes acute hepatotoxicity by lipid peroxidation and oxidative DNA damage, and induces liver carcinoma in humans and laboratory animals. This study was performed to examine the protective effects of KRG against hepatotoxicity induced by $AFB_1$ using liver-specific serum marker analysis, histopathology, and terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling. In addition, to elucidate the possible mechanism of hepatoprotective effects, superoxide dismutase, catalase, glutathione peroxidase, and malondialdehyde were analyzed. Rats were treated with 250 mg/kg of KRG (KRG group) or saline ($AFB_1$ group) for 4 weeks and then received 150 ${\mu}g/kg$ of $AFB_1$ intraperitoneally for 3 days. Rats were sacrificed at 12 h, 24 h, 48 h, 72 h, or 1 wk after $AFB_1$ treatment. In the KRG pre-treatment group, serum alanine aminotransferase, aspartate aminotransferase, and malondialdehyde levels were low, but superoxide dismutase, catalase, and glutathione peroxidase activities were high as compared to the $AFB_1$ alone group. Histopathologically, $AFB_1$ treatment induced necrosis and apoptosis in hepatocytes, and led to inflammatory cells infiltration in the liver. KRG pre-treatment ameliorated these changes. These results indicate that KRG may have protective effects against hepatotoxicity induced by $AFB_1$ that involve the antioxidant properties of KRG.

Microbial Biodegradation and Toxicity of Vinclozolin and its Toxic Metabolite 3,5-Dichloroaniline

  • Lee, Jung-Bok;Sohn, Ho-Yong;Shin, Kee-Sun;Kim, Jong-Sik;Jo, Min-Sub;Jeon, Chun-Pyo;Jang, Jong-Ok;Kim, Jang-Eok;Kwon, Gi-Seok
    • Journal of Microbiology and Biotechnology
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    • v.18 no.2
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    • pp.343-349
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    • 2008
  • Vinclozolin, an endocrine disrupting chemical, is a chlorinated fungicide widely used to control fungal diseases. However, its metabolite 3,5-dichloroaniline is more toxic and persistent than the parent vinclozolin. For the biodegradation of vinclozolin, vinclozolin- and/or 3,5-dichloroaniline-degrading bacteria were isolated from pesticide-polluted agriculture soil. Among the isolated bacteria, a Rhodococcus sp. was identified from a 16S rDNA sequence analysis and named Rhodococcus sp. T1-1. The degradation ratios for vinclozolin or 3,5-dichloroaniline in a minimal medium containing vinclozolin $(200{\mu}ml)$ or 3,5-dichloroaniline $(120{\mu}g/ml)$ were 90% and 84.1%, respectively. Moreover, Rhodococcus sp. T1-1 also showed an effective capability to biodegrade dichloroaniline isomers on enrichment cultures in which they were contained. Therefore, these results suggest that Rhodococcus sp. T1-1 can bioremediate vinclozolin as well as 3,5-dichloroaniline.

Effect of Grape Skin Extract on Apoptosis in MDA-MB-231 Human Breast Cancer Cells (포도껍질 추출물이 인체유방암세포 사멸에 미치는 영향)

  • Park, Min-A;Kang, Keum-Jee
    • Journal of the East Asian Society of Dietary Life
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    • v.25 no.1
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    • pp.87-98
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    • 2015
  • We investigated the apoptotic effects of grape skin extracts (GSE) and related gene expressions in human breast cancer MDA-MB-231 cells cultured in the presence of 0, 0.5, 1 and 1.5 mg/mL of GSE for 72 hours. MTT assay, trypan blue and nuclei staining showed lower cellular mitochondrial activities and increased cell deaths with a higher concentration of GSE (p<0.05). Increased cell number with fragmentated DNA of sub-G1 phase was calculated as a measure of apoptotic cell death by FACS analysis (p<0.05). In particular, apoptotic cell death caused markedly increased in the 1 and 1.5 mg/mL of GSE groups, as revealed by flow cytometry (Annexin V-FITC). RT-PCR analysis was performed on apoptotic and preapoptotic genes. Expression of the apoptosis suppressor gene bcl-2 significantly decreased, proapoptotic gene bax was significantly increased and procaspase-3 showing the presence of caspase-3 significantly decreased (p<0.05). Furthermore, bcl-2/bax ratio which is considered to be an important indicator of apoptosis, significantly decreased in a concentration-dependent manner (p<0.05). These results indicated that GSE induces apoptosis in MDA-MB-231 human breast cancer cells.

Esculetin Induces Apoptosis through Caspase-3 Activation in Human Leukemia U937 Cells (Esculetin의 caspase-3 활성을 통한 U937 인체 혈구암세포의 세포사멸 유도)

  • Park, Cheol;Hyun, Sook-Kyung;Shin, Woo-Jin;Chung, Kyung-Tae;Choi, Byung-Tae;Kwon, Hyun-Ju;Hwang, Hye-Jin;Kim, Byung-Woo;Park, Dong-Il;Lee, Won-Ho;Choi, Yung-Hyun
    • Journal of Life Science
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    • v.19 no.2
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    • pp.249-255
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    • 2009
  • Esculetin, a coumarin compound, has been known to inhibit proliferation and induce apoptosis in several types of human cancer cells. However, the molecular mechanisms involved in esculetin-induced apoptosis are still uncharacterized in human leukemia cells. In this study, we have investigated whether esculetin exerts anti-proliferative and apoptotic effects on human leukemia U937 cells. It was found that esculetin could inhibit cell viability in a time-dependent manner, which was associated with the induction of apoptotic cell death such as increased populations of apoptotic- sub G1 phase. Apoptosis of U937 cells by esculetin was associated with an inhibition of Bcl-2/Bax binding activity, formation of tBid, down-regulation of X-linked inhibitor of apoptotic protein (XIAP) expression, and up-regulation of death receptor 4 (DR4) and FasL expression. Esculetin treatment also induced the degradation of ${\beta}$-catenin and DNA fragmentation factor 45/inhibitor of caspase-activated DNase (DFF45/ICAD). Furthermore, a caspase-3 specific inhibitor, z-DEVD-fmk, significantly inhibited sub-G1 phase DNA content, morphological changes and degradation of ${\beta}$-catenin and DEE45/ICAD. These results indicated that a key regulator in esculetin-induced apoptosis was caspase-3 in human leukemia U937 cells.