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8-60hIPP5m-Induced G2/M Cell Cycle Arrest Involves Activation of ATM/p53/p21cip1/waf1 Pathways and Delayed Cyclin B1 Nuclear Translocation

  • Zeng, Qi-Yan;Zeng, Lin-Jie;Huang, Yu;Huang, Yong-Qi;Zhu, Qi-Fang;Liao, Zhi-Hong
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.9
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    • pp.4101-4107
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    • 2014
  • Protein phosphatase 1 (PP1) is a major serine/threonine phosphatase that controls gene expression and cell cycle progression. The active mutant IPP5 ($8-60hIPP5^m$), the latest member of the inhibitory molecules for PP1, has been shown to inhibit the growth of human cervix carcinoma cells (HeLa). In order to elucidate the underlying mechanisms, the present study assessed overexpression of $8-60hIPP5^m$ in HeLa cells. Flow cytometric and biochemical analyses showed that overexpression of $8-60hIPP5^m$ induced G2/M-phase arrest, which was accompanied by the upregulation of cyclin B1 and phosphorylation of G2/M-phase proteins ATM, p53, $p21^{cip1/waf1}$ and Cdc2, suggesting that $8-60hIPP5^m$ induces G2/M arrest through activation of the ATM/p53/$p21^{cip1/waf1}$/Cdc2/cyclin B1 pathways. We further showed that overexpression of $8-60hIPP5^m$ led to delayed nuclear translocation of cyclin B1. $8-60hIPP5^m$ also could translocate to the nucleus in G2/M phase and interact with $pp1{\alpha}$ and Cdc2 as demonstrated by co-precipitation assay. Taken together, our data demonstrate a novel role for $8-60hIPP5^m$ in regulation of cell cycle in HeLa cells, possibly contributing to the development of new therapeutic strategies for cervix carcinoma.

Effects of Oocyte Maturational Age and Activation Conditions on the Development of Porcine Parthenogenetic Embryos

  • Kwon, Dae-Jin;Park, Joo-Hee;Park, Choon-Keun;Yang, Boo-Keun;Cheong, Hee-Tae
    • Reproductive and Developmental Biology
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    • v.31 no.2
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    • pp.77-82
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    • 2007
  • This study was conducted to investigate the effects of oocyte maturational age and activation condition on in vitro development of porcine parthenogenetic embryos (parthenotes). Porcine follicular oocytes were matured in vitro for 30 to 44 hr. Maturation rate was examined during in vitro maturation (IVM) every 2 hr interval. The cdc2 kinase activity was measured at 36 and 44 hr of IVM. Some oocytes were activated at 36 or 44 hr of IVM by three different conditions; 1) single electric stimulation (1.5 kV/cm for $30{\mu}sec$; ES), 2) double electric stimulations (1.5 kV/cm for $30{\mu}sec$, followed by 1.0 kV/cm for $50{\mu}sec$ after 1 hr; ES+ES) or 3) ES+ES followed by culture in 6-dimethlyaminopurine (6-DMAP) for 4 hr (ES+ES+D), and cultured for 6-7 days. Maturation rate was significantly increased as culture period was increased to 36 hr (66.9%, p<0.05), and then gradually increased to 87.1% at 44 hr of IVM. The cdc2 kinase activity was decreased (p<0.05) with culture period prolonged from 36 hr to 44 hr. Lower blastocyst formation rate (4.3%, p<0.05) were obtained by ES in 36 hr-matured oocytes compared to other treatments (16.5 and 20.5%) in the same age and the same treatment in 44 hr-matured oocytes (15.0%). High blastocyst formation rate (23.6%) was obtained by ES+ES+D in 44 hr-matured oocytes (p<0.05). These results demonstrate that porcine oocyte activation and in vitro development of parthenotes can be affected by interactions between oocyte maturational age and activation condition.

Therapeutic Effect of Bogijetongtanggammi-bang on Peripheral Nerve Injury (보기제통탕감미방(補氣除痛湯減味方)의 랫드 말초신경 손상에 대한 회복 효과 연구)

  • Kim, Jin-Mi;Cho, Chung-Sik;Kim, Chul-Jung
    • The Journal of Internal Korean Medicine
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    • v.33 no.1
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    • pp.83-101
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    • 2012
  • Objectives : This study was aimed to investigate the therapeutic effect of Bogijetongtanggammi-bang (BJTG) on injury of the peripheral nerve tissues. Methods : Rats were divided into 2 groups. The rats of the first group were injected with Taxol (1.25 mg/kg) to their sciatic nerves, once each. The sciatic nerves of the rats of the second group were crushed by forcept for 30 seconds. Rats were administered with BJTG (400 mg/kg) or 0.9% saline for 5 days. Changes of DRG neurons, Schwann cells, Cdc2, caspase 3. phospho-p44/42 Erk1/2, phospho-vimentin and ${\beta}1$ integrin were observed by fluorescent microscope and analysed in western blot. Results : In Taxol-treated SD rat models, BJTG up-regulated neurite outgrowth, Schwann cells, Cdc2 and phospho-Erk1/2, and down-regulated caspase 3. In pressure-injured rat models, BJTG up-regulated axons of sciatic nerve, Schwann cells, Cdc2, phospho-vimentin, ${\beta}1$ integrin, and down-regulated caspase 3. Conclusions : Taken together, BJTG was promotive of nerve regeneration on SNI as well as Taxol-induced nerve injury. BJTG had a pharmaceutical property enhancing recovery of injured peripheral nerves and could be a candidate for drug development after further research.

Transition over 35 Years in the Incidence Rates of Primary Central Nervous System Tumors in Shanghai, China and Histological Subtyping Based on a Single Center Experience Spanning 60 Years

  • Shen, Fang;Wu, Chun-Xiao;Yao, Yu;Peng, Peng;Qin, Zhi-Yong;Wang, Yin;Zheng, Ying;Zhou, Liang-Fu
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.12
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    • pp.7385-7393
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    • 2013
  • Background: Only few epidemiological data on primary central nervous system (CNS) tumors in Shanghai have been reported. Methods: All cases of primary CNS tumors that were registered at Center for Disease Control and Prevention (CDC) were collected (1973-2007: urban Shanghai; 2003-2007: whole Shanghai city). Trends were analyzed using joinpoint analysis and rates were stratified by age, gender and region. Histological data were collected from both CDC and Huashan Hospital. Results: From 1973 to 2007, the five-year average incidence rate in urban Shanghai increased in both genders, especially in the elderly population. Joinpoint analysis showed the age-adjusted incidence rate for males increased first but then plateaued, whilst rates for females continued increasing over the 35 years. For the five-year status quo (2003-2007), rural had a higher age-adjusted incidence rate than urban populations, and females higher than males, especially those with advanced age. According to CDC (2003-2007) and Huashan Hospital (1951-2011), the two most common histological subtypes were neuroepithelial tumors (with male predominance) and meningiomas (with female predominance). Conclusions: In Shanghai, a steadily increased incidence rate of primary CNS tumors was observed in general, and in the elderly and female population in particular.

Growth Promoting Effects of Oriental Medicinal Drugs on Sciatic Nerve Regeneration in the Rat

  • Jo Hyun-Kyung;NamGung Uk;Seol In-Chan;Kim Yoon-Sik
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.19 no.6
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    • pp.1666-1672
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    • 2005
  • Oriental medicinal drugs have a broad spectrum of clinical use for the cure of nervous system diseases including brain ischemic damages or neuropathies. Yet, specific drugs or drug components used in the oriental medicine in relation to none fiber regeneration are not known. In the present study, possible growth promoting effects of oriental medicinal drugs were investigated in the injured sciatic nerve system in the rat. By immunofluorescence staining, we found that Jahageo (JHG, Hominis placenta) increased Induction levels of axonal growth associated protein GAP-43 in the rat sciatic none. Small growth promoting activity was found in Golsebo (GSB, Drynariae rhizoma) and Baikhasuo (BHSO, Polygoni multiflori radix) drugs. JHG also increased cell cycle protein Cdc2 levels in the injured area of the sciatic nerves. Immunofluorescence staining indicated that induced Cdc2 protein was mostly localized in the Schwann cells in the injury area, implying that JHG activity might be related to increased Schwann cell proliferation during axonal regeneration. Moreover, levels of phospho-extracellular signal-regulated (ERK) pathway in the injured neNes were elevated by JHG treatment while levels of total ERK were unaltered. In vivo measurement of axonal regeneration using retrograde tracer showed that JHG, GSB and BHSO significantly enhanced Dil-labeled regenerating motor neurons compared with saline control. The present data suggest that oriental medicinal drugs such as JHG, GSB, and BHSO may be a useful target for developing specific drugs of axonal regeneration.

Induction of G1 Arrest by Methanol Extract of Lycopus lucidus in Human Lung Adenocarcinoma A549 Cells (택란 메탄올 추출물에 의한 인체 폐암 세포주 A549의 G1 arrest 유발)

  • Park, Hyun-Jin;Jin, Soojung;Oh, You Na;Yun, Seung-Geun;Lee, Ji-Young;Kwon, Hyun Ju;Kim, Byung Woo
    • Journal of Life Science
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    • v.23 no.9
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    • pp.1109-1117
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    • 2013
  • Induction of G1 Arrest by Methanol Extract of Lycopus lucidus in Human Lung Adenocarcinoma A549 Cells Lycopus lucidus, a herbaceous perennial, is used as a traditional remedy in East Asia, including China and Korea. It has been reported that L. lucidus has anti-allergic effects, inhibitory effects on cholesterol acyltransferase in high glucose-induced vascular inflammation, and anti-proliferative effects in human breast cancer cells. However, the molecular mechanisms of the anti-cancer effects of L. lucidus have not yet been fully determined. In this study, we evaluated the anti-cancer effect and the mechanism of action of L. lucidus in human lung adenocarcinoma A549 cells using methanol extracts of L. lucidus (MELL). MELL treatment showed cytotoxic activity in a dose-dependent manner and induced G1 arrest in A549 cells. The induction of G1 arrest by MELL was associated with the up-regulation of phospho-CHK2 and the down-regulation of Cdc25A phosphatase. In addition, MELL treatment induced decreased expression of G1/S transition-related proteins, including CDK2, CDK4, CDK6, cyclin D1 and cyclin E. MELL also regulated the mRNA expression of CDK2 and cyclin E. On the other hand, the expression of p53 and the cyclin-dependent kinase inhibitor p21 was not induced by MELL. Collectively, these results suggest that MELL may exert an anti-cancer effect by cell cycle arrest at G1 phase through the ATM/CHK2/Cdc25A/CDK2 pathway in A549 cells.

A Cross Sectional Study of Kretek Smoking in Indonesia as a Major Risk to Public Health

  • Palipudi, Krishna;Mbulo, Lazarous;Kosen, Soewarte;Tjandra, Aditama;Kadarmanto, Kadarmanto;Qureshi, Farukh;Andes, Linda;Sinha, Dhirendra N;Asma, Samira
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.16
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    • pp.6883-6888
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    • 2015
  • Background: Tobacco smoking is a major cause of preventable disease and death worldwide. Kreteks are clove-flavored cigarettes made from a combination of tobacco and ground-clove mixed with a sauce, smoked widely in Indonesia. Because health and social consequences of kretek smoking are potentially as great as those of traditional cigarettes, this study examines the prevalence of kretek smoking in Indonesia and associated risk factors. Materials and Methods: The study used nationally representative Indonesia Global Adult Tobacco Survey data. Multiple logistic regression analysis was employed to identify correlates of kretek smoking. Results: One-third of Indonesian adults smoked tobacco of which about 90.0% smoked kreteks. Prevalence of kretek smoking among men (60.9%) was more than 25 times the rate among women (2.3%). Overall, the highest prevalence of kretek use was in the age group 45-54 years (36.5%), followed by 34-44 (35.1%), 25-34 (34.2 %), and 55-64 years (32.8%). By wealth index, prevalence of kreteks smoking among those in the middle index was almost 50% above the rate for the wealthiest group (36.4% vs 24.8% respectively). Logistic regression results showed that being male, being older, having less education, and being less wealthy were significant predictors of kretek smoking, while urban vs rural residence was not. Conclusions: Kretek smoking is common in Indonesia and is entrenched in the sociocultural fabric of the country. However, potential consequences of kretek smoking, particularly as risks for noncommunicable diseases, underscore the importance of a comprehensive approach to tobacco control as outlined in the World Health Organization's MPOWER strategies.

Generation of 1E8 Single Chain Fv-Fc Construct Against Human CD59

  • Hong, Jeong-Won;Cho, Woon-Dong;Hong, Kwon-Pyo;Kim, So-Seul;Son, Seung-Myoung;Yun, Seok-Joong;Lee, Ho-Chang;Yoon, Sang-Soon;Song, Hyung-Geun
    • IMMUNE NETWORK
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    • v.12 no.1
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    • pp.33-39
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    • 2012
  • Background: Therapeutic approaches using monoclonal antibodies (mAbs) against complement regulatory proteins (CRPs:i.e.,CD46,CD55 and CD59) have been reported for adjuvant cancer therapy. In this study, we generated a recombinant 1E8 single-chain anti-CD59 antibody (scFv-Fc) and tested anti-cancer effect.by using complement dependent cytotoxicity (CDC). Methods: We isolated mRNA from 1E8 hybridoma cells and amplified the variable regions of the heavy chain (VH) and light chain (VL) genes using reversetranscriptase polymerase chain reaction (RT-PCR). Using a linker, the amplified sequences for the heavy and light chains were each connected to the sequence for a single polypeptide chain that was designed to be expressed. The VL and VH fragments were cloned into the pOptiVEC-TOPO vector that contained the human CH2-CH3 fragment. Then, 293T cells were transfected with the 1E8 single-chain Fv-Fc (scFv-Fc) constructs. CD59 expression was evaluated in the prostate cancer cell lines using flow cytometry. The enhancement of CDC effect by mouse 1E8 and 1E8 scFv-Fc were evaluated using a cytotoxicity assay. Results: The scFv-Fc constructs were expressed by the transfected 293T cells and secreted into the culture medium. The immunoreactivity of the secreted scFv-Fc construct was similar to that of the mouse 1E8 for CCRF-CEM cells. The molecular masses of 1E8 scFv-Fc were about 120 kDa and 55 kDa under reducing and non-reducing conditions, respectively. The DNA sequence of 1E8 scFv-Fc was obtained and presented. CD59 was highly expressed by the prostate cancer cell line. The recombinant 1E8 scFv-Fc mAb revealed significantly enhanced CDC effect similar with mouse 1E8 for prostate cancer cells. Conclusion: A 1E8 scFv-Fc construct for adjuvant cancer therapy was developed.