• Title/Summary/Keyword: Interference regulation

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HDAC6 siRNA Inhibits Proliferation and Induces Apoptosis of HeLa Cells and its Related Molecular Mechanism

  • Qin, Hai-Xia;Cui, Hong-Kai;Pan, Ying;Yang, Jun;Ren, Yan-Fang;Hua, Cai-Hong;Hua, Fang-Fang;Qiao, Yu-Huan
    • Asian Pacific Journal of Cancer Prevention
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    • v.13 no.7
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    • pp.3367-3371
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    • 2012
  • Objective: To investigate the effects of histone deacetylase 6 (HDAC6) siRNA on cell proliferation and cell apoptosis of the HeLa cervical carcinoma cell line and the molecular mechanisms involved. Methods: Division was into three groups: A, the untreated group; B, the control siRNA group; and C, the HDAC6 siRNA group. Lipofectamine 2000 was used for siRNA transfection, and Western blot analysis was used to determine the protein levels. Cell proliferation and apoptosis were characterized using a CCK-8 assay and flow cytometry, respectively. Results: HDAC6 protein expression in the HDAC6 siRNA-transfection group was significantly lower (P < 0.05) than in the untreated and control siRNA groups. The CCK-8 kit results demonstrated that the proliferation of HeLa cells was clearly inhibited in the HDAC6 siRNA transfection group (P < 0.05). In addition, flow cytometry revealed that the early apoptotic rate ($26.0%{\pm}0.87%$) was significantly elevated (P < 0.05) as compared with the untreated group ($10.6%{\pm}1.19%$) and control siRNA group ($8.61%{\pm}0.98%$). Furthermore, Western blot analysis indicated that bcl-2 protein expression in the HDAC6 siRNA-transfection group was down-regulated, whereas the expression of p21 and bax was up-regulated. Conclusion: HDAC6 plays an essential role in the occurrence and development of cervical carcinoma, and the down-regulation of HDAC6 expression may be useful molecular therapeutic method.

In vitro Study of Nucleostemin as a Potential Therapeutic Target in Human Breast Carcinoma SKBR-3 Cells

  • Guo, Yu;Liao, Ya-Ping;Zhang, Ding;Xu, Li-Sha;Li, Na;Guan, Wei-Jun;Liu, Chang-Qing
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.5
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    • pp.2291-2295
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    • 2014
  • Although nucleolar protein nucleostemin (NS) is essential for cell proliferation and early embryogenesis and expression has been observed in some types of human cancer and stem cells, the molecular mechanisms involved in mediation of cell proliferation and cell cycling remains largely elusive. The aim of the present study was to evaluate NS as a potential target for gene therapy of human breast carcinoma by investigating NS gene expression and its effects on SKBR-3 cell proliferation and apoptosis. NS mRNA and protein were both found to be highly expressed in all detected cancer cell lines. The apoptotic rate of the pcDNA3.1-NS-Silencer group ($12.1-15.4{\pm}3.8%$) was significantly higher than those of pcDNA3.1-NS ($7.2-12.0{\pm}1.7%$) and non-transfection groups ($4.1-6.5{\pm}1.8%$, P<0.01). MTT assays showed the knockdown of NS expression reduced the proliferation rate of SKBR-3 cells significantly. Matrigel invasion and wound healing assays indicated that the number of invading cells was significantly decreased in the pcDNA3.1-NS-siRNA group (P<0.01), but there were no significant difference between non-transfected and over-expression groups (P>0.05). Moreover, RNAi-mediated NS down-regulation induced SKBR-3 cell G1 phase arrest, inhibited cell proliferation, and promoted p53 pathway-mediated cell apoptosis in SKBR-3 cells. NS might thus be an important regulator in the G2/M check point of cell cycle, blocking SKBR-3 cell progression through the G1/S phase. On the whole, these results suggest NS might be a tumor suppressor and important therapeutic target in human cancers.

UHRF2 mRNA Expression is Low in Malignant Glioma but Silencing Inhibits the Growth of U251 Glioma Cells in vitro

  • Wu, Ting-Feng;Zhang, Wei;Su, Zuo-Peng;Chen, San-Song;Chen, Gui-Lin;Wei, Yong-Xin;Sun, Ting;Xie, Xue-Shun;Li, Bin;Zhou, You-Xin;Du, Zi-Wei
    • Asian Pacific Journal of Cancer Prevention
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    • v.13 no.10
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    • pp.5137-5142
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    • 2012
  • UHRF2 is a member of the ubiquitin plant homeo domain RING finger family, which has been proven to be frequently up-regulated in colorectal cancer cells and play a role as an oncogene in breast cancer cells. However, the role of UHRF2 in glioma cells remains unclear. In this study, we performed real-time quantitative PCR on 32 pathologically confirmed glioma samples (grade I, 4 cases; grade II, 11 cases; grade III, 10 cases; and grade IV, 7 cases; according to the 2007 WHO classification system) and four glioma cell lines (A172, U251, U373, and U87). The expression of UHRF2 mRNA was significantly lower in the grade III and grade IV groups compared with the noncancerous brain tissue group, whereas its expression was high in A172, U251, and U373 glioma cell lines. An in vitro assay was performed to investigate the functions of UHRF2. Using a lentivirus-based RNA interference (RNAi) approach, we down-regulated UHRF2 expression in the U251 glioma cell line. This down-regulation led to the inhibition of cell proliferation, an increase in cell apoptosis, and a change of cell cycle distribution, in which S stage cells decreased and G2/M stage cells increased. Our results suggest that UHRF2 may be closely related to tumorigenesis and the development of gliomas.

Variation of Amylose Content Using dsRNAi Vector by Targeting 3'-UTR Region of GBSSI Gene in Rice (GBSSI 유전자 3'UTR 영역의 발현 억제 dsRNAi 벡터를 이용한 아밀로스함량 조절 벼 개발)

  • Park, Hyang-Mi;Choi, Man-Soo;Chun, Areum;Lee, Jeung-Heui;Kim, Myeong-Ki;Kim, Yeon-Gyu;Shin, Dong-Bum;Lee, Jang-Yong;Kim, Yul-Ho
    • Korean Journal of Breeding Science
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    • v.42 no.5
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    • pp.515-524
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    • 2010
  • The amylose content of starch is a major factor in the texture of cooked cereal grains. Therefore, down-regulation of amylose synthesis is one of the alternative method to improve eating quality of rice. We developed transgenic rice plants designed to suppress granule-bound starch synthase I(GBSSI) gene using RNA interference(RNAi) technology. Transgenic plants with RNAi vector containing the 3'-UTR region of GBSSI showed a lower amylose content in rice endosperm than that of wild-type. The range of amylose content was 5.9~9.0% in the transgenic plants, whereas that of wild-type was 17.7~18.0%. Transgenic rices showed the decrease of short chain and the increase of long chain by analyzing chain length distribution of amylopectin in the endosperm. In the SEM micrographs, we found that compound starch granules in whole grains of the wild-type rice were readily split during fracturing, while the starch granules in RNAi-transgenic lines showed small voluminous, non-angular rounded bodies.

Changes in the Riverbed Landforms Due to the Artificial Regulation of Water Level in the Yeongsan River (인위적인 보 수위조절로 인한 영산강 하도 지형 변화)

  • Lim, Young Shin;Kim, Jin Kwan
    • Journal of The Geomorphological Association of Korea
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    • v.27 no.1
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    • pp.1-19
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    • 2020
  • A river bed which is submerged in water at high flow and becomes part of the river at low flow, serves as a bridge between the river and the land. The channel bar creates a unique ecosystem with vegetation adapted to the particular environment and the water pool forms a wetland that plays a very important role in the environment. To evaluate anthropogenic impacts on the river bed in the Middle Yeongsangang River, the fluvial landforms in the stream channel were analyzed using multi-temporal remotely-sensed images. In the aerial photograph of 2005 taken before the construction of the large weirs, oxbow lakes, mid-channel bars, point bars, and natural wetlands between the artificial levees were identified. Multiple bars divided the flow of stream water to cause the braided pattern in a particular section. After the construction of the Seungchon weir, aerial photographs of 2013 and 2015 revealed that most of the fluvial landforms disappeared due to the dredging of its riverbed and water level control(maintenance at 7.5El.m). Sentinel-2 images were analyzed to identify differences between before and after the opening of weir gate. Change detection was performed with the near infrared and shortwave infrared spectral bands to effectively distinguish water surfaces from land. As a result, water surface area of the main stream of the Yeongsangang River decreased by 40% from 1.144km2 to 0.692km2. A large mid-channel bar that has been deposited upstream of the weir was exposed during low water levels, which shows the obvious influence of weir on the river bed. Newly formed unvegetated point bars that were deposited on the inside of a meander bend were identified from the remotely sensed images. As the maintenance period of the weir gate opening was extended, various habitats were created by creating pools and riffles around the channel bars. Considering the ecological and hydrological functions of the river bed, it is expected that the increase in bar areas through weir gate opening will reduce the artificial interference effect of the weir.

MiR-188-5p regulates the proliferation and differentiation of goat skeletal muscle satellite cells by targeting calcium/calmodulin dependent protein kinase II beta

  • Jing Jing;Sihuan Zhang;Jinbo Wei;Yuhang Yang;Qi Zheng;Cuiyun Zhu;Shuang Li;Hongguo Cao;Fugui Fang;Yong Liu;Ying-hui Ling
    • Animal Bioscience
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    • v.36 no.12
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    • pp.1775-1784
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    • 2023
  • Objective: The aim of this study was to reveal the role and regulatory mechanism of miR-188-5p in the proliferation and differentiation of goat muscle satellite cells. Methods: Goat skeletal muscle satellite cells isolated in the pre-laboratory were used as the test material. First, the expression of miR-188-5p in goat muscle tissues at different developmental stages was detected by quantitative reverse transcription polymerase chain reaction (qRT-PCR). In addition, miR-188-5p was transfected into goat skeletal muscle satellite cells by constructing mimics and inhibitors of miR-188-5p, respectively. The changes of differentiation marker gene expression were detected by qPCR method. Results: It was highly expressed in adult goat latissimus dorsi and leg muscles, goat fetal skeletal muscle, and at the differentiation stage of muscle satellite cells. Overexpression and interference of miR-188-5p showed that miR-188-5p inhibited the proliferation and promoted the differentiation of goat muscle satellite cells. Target gene prediction and dual luciferase assays showed that miR-188-5p could target the 3'untranslated region of the calcium/calmodulin dependent protein kinase II beta (CAMK2B) gene and inhibit luciferase activity. Further functional studies revealed that CAMK2B promoted the proliferation and inhibited the differentiation of goat muscle satellite cells, whereas si-CAMK2B restored the function of miR-188-5p inhibitor. Conclusion: These results suggest that miR-188-5p inhibits the proliferation and promotes the differentiation of goat muscle satellite cells by targeting CAMK2B. This study will provide a theoretical reference for future studies on the molecular mechanisms of skeletal muscle development in goats.

The role of RNA epigenetic modification-related genes in the immune response of cattle to mastitis induced by Staphylococcus aureus

  • Yue Xing;Yongjie Tang;Quanzhen Chen;Siqian Chen;Wenlong Li;Siyuan Mi;Ying Yu
    • Animal Bioscience
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    • v.37 no.7
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    • pp.1141-1155
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    • 2024
  • Objective: RNA epigenetic modifications play an important role in regulating immune response of mammals. Bovine mastitis induced by Staphylococcus aureus (S. aureus) is a threat to the health of dairy cattle. There are numerous RNA modifications, and how these modification-associated enzymes systematically coordinate their immunomodulatory effects during bovine mastitis is not well reported. Therefore, the role of common RNA modification-related genes (RMRGs) in bovine S. aureus mastitis was investigated in this study. Methods: In total, 80 RMRGs were selected for this study. Four public RNA-seq data sets about bovine S. aureus mastitis were collected and one additional RNA-seq data set was generated by this study. Firstly, quantitative trait locus (QTL) database, transcriptome-wide association studies (TWAS) database and differential expression analyses were employed to characterize the potential functions of selected enzyme genes in bovine S. aureus mastitis. Correlation analysis and weighted gene co-expression network analysis (WGCNA) were used to further investigate the relationships of RMRGs from different types at the mRNA expression level. Interference experiments targeting the m6 A demethylase FTO and utilizing public MeRIP-seq dataset from bovine Mac-T cells were used to investigate the potential interaction mechanisms among various RNA modifications. Results: Bovine QTL and TWAS database in cattle revealed associations between RMRGs and immune-related complex traits. S. aureus challenged and control groups were effectively distinguished by principal component analysis based on the expression of selected RMRGs. WGCNA and correlation analysis identified modules grouping different RMRGs, with highly correlated mRNA expression. The m6 A modification gene FTO showed significant effects on the expression of m6 A and other RMRGs (such as NSUN2, CPSF2, and METTLE), indicating complex co-expression relationships among different RNA modifications in the regulation of bovine S. aureus mastitis. Conclusion: RNA epigenetic modification genes play important immunoregulatory roles in bovine S. aureus mastitis, and there are extensive interactions of mRNA expression among different RMRGs. It is necessary to investigate the interactions between RNA modification genes regulating complex traits in the future.

The moderating effects of ego-resilience on the effects of parents' child-rearing attitude perceived by adolescents and school life adaptation on problem behavior (청소년이 지각한 부모의 양육태도와 학교생활적응이 문제행동에 미치는 영향에서 자아탄력성의 조절효과)

  • Kim, Ji Hye;Yu, Nan Sook
    • Journal of Korean Home Economics Education Association
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    • v.31 no.1
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    • pp.1-19
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    • 2019
  • The purposes of this study were to concretely reveal the effect of the parents' child-rearing attitude perceived by adolescents and the school life adaptation on the problem behavior, and to verify the moderating effect of the ego-resilience on the relationship between the parents' child-rearing attitude and the school life adaptation and the adolescent problem behavior. This study analyzed a total of 2,107 students in the first year of high school, which was the 4th year data(2013) of Korea Children Youth Panel Survey(KCYPS) 2010. The reliability, descriptive statistics, t-test, and hierarchical regression analysis were conducted using SPSS WIN 22.0. The results were as follows. First, the effect of the parents' child-rearing attitude(supervision, affection, reasonable explanation, excessive interference, excessive expectation, and inconsistency), school life adaptation(relationship with teacher, relationship with friend, school regulation, and learning activity), and ego-resilience on the adolescent problem behavior was analyzed. As a result, the relationship with friend(-) had the highest influence on the adolescent problem behavior, followed by learning activity(-), inconsistency(+), ego-resilience(-), excessive interference(+), and supervision(-). However, the remaining sub-variables did not have any significant influence on the adolescent problem behavior. Second, the moderating effect of the ego-resilience on the relationship among the parents' child-rearing attitude, adaptation to school life, and adolescent problem behavior. The ego-resilience was found to moderate the effects of parents' positive child-rearing attitude, interpersonal relationships, and school adaptation on the adolescent problem behavior. However, the moderating effect was not significant for the effect of negative child-rearing attitude on the adolescent problem behavior. Therefore, various ego-resilience enhancement programs need to be developed and researched as a part of the safety education through the home economics class.

Standards of Protection in Investment Arbitration for Upcoming Climate Change Cases (기후변화 관련 사건에 적용되는 국제투자중재의 투자자 보호 기준)

  • Kim, Dae-Jung
    • Journal of Arbitration Studies
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    • v.24 no.2
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    • pp.33-52
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    • 2014
  • Although climate change is a global scale question, some concerns have been raised that principles of investment arbitration may not adequately address the domestic implementation of climate change measures. A recent ICSID investment arbitration of Vattenfall v. Germany with regard to the investor's alleged damages from the phase-out of nuclear plants is a salient climate change case. The 2005 Kyoto Protocol was made to reduce greenhouse gas emissions and it provides a number of flexible mechanisms such as Joint Implementation (JI) and Clean Development Mechanism (CDM). Implementation of the Kyoto Protocol allows dispute settlement through investor-state arbitration. Any initiation of stricter emission standards can violate the prohibition on expropriations in investment agreements, regardless of the measures created to reduce greenhouse gas emissions. The effect-based expropriation doctrine can charge changes to existing emission standards as interference with the use of property that goes against the legitimate expectation of a foreign investor. In regulatory chill, threat of investor claims against the host state may preclude the strengthening of climate change measures. Stabilization clauses also have a freezing effect on the hosting state's regulation and a new law applicable to the investment. In the fair and equitable standard, basic expectations of investors when entering into earlier carbon-intensive operations can be affected by a regulation seeking to change into a low-carbon approach. As seen in the Methanex tribunal, a non-discriminatory and public purpose of environmental protection measures should be considered as non-expropriation in the arbitral tribunal unless its decision would intentionally impede a foreign investor's investment.

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Study on the screening method for determination of heavy metals in cellular phone for the restrictions on the use of certain hazardous substances (RoHS) (유해물질 규제법(RoHS)에 따른 휴대폰 내의 중금속 함유량 측정을 위한 스크리닝법 연구)

  • Kim, Y.H.;Lee, J.S.;Lim, H.B.
    • Analytical Science and Technology
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    • v.23 no.1
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    • pp.1-14
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    • 2010
  • It is of importance that all countries in worldwide, including EU and China, have adopted the Restrictions on the use of certain Hazardous Substances (RoHS) for all electronics. IEC62321 document, which was published by the International Electronics Committee (IEC) can have conflicts with the standards in the market. On the contrary Publicly Accessible Specification (PAS) for sampling published by IEC TC111 can be adopted for complementary application. In this work, we tried to find a route to disassemble and disjoint cellular phone sample, based on PAS and compare the screening methods available in the market. For this work, the cellular phone produced in 2001, before the regulation was born, was chosen for better detection. Although X-ray Fluorescence (XRF) showed excellent performance for screening, fast and easy handling, it can give information on the surface, not the bulk, and have some limitations due to significant matrix interference and lack of variety of standards for quantification. It means that screening with XRF sometimes requires supplementary tool. There are several techniques available in the market of analytical instruments. Laser ablation (LA) ICP-MS, energy dispersive (ED) XRF and scanning electron microscope (SEM)-energy dispersive X-ray (EDX) were demonstrated for screening a cellular phone. For quantitative determination, graphite furnace atomic absorption spectrometry (GF-AAS) was employed. Experimental results for Pb in a battery showed large difference in analytical results in between XRF and GF-AAS, i.e., 0.92% and 5.67%, respectively. In addition, the standard deviation of XRF was extremely large in the range of 23-168%, compared with that in the range of 1.9-92.3% for LA-ICP-MS. In conclusion, GF-AAS was required for quantitative analysis although EDX was used for screening. In this work, it was proved that LA-ICP-MS can be used as a screening method for fast analysis to determine hazardous elements in electrical products.