Functional expression of mechanosensitive two-pore domain potassium channel in human bladder carcinoma cells

Yangmi Kim, Kyung Sun Park 충북대학교 동물의학연구소 2013 Journal of Biomedical and Translational Research Vol.14 No.2

Bladder cancer is a common cancer in smoking men and may correlate with mechanosensitive potassium channels because the urinary bladder is a stretch sensing organ. Two-pore K+ channels (K2P), such as TASK3 and TREK1, have recently been shown to play a critical role in both cell apoptosis and tumorigenesis. Of the channels, TREK1 can be activated by many physiological stimuli, including polyunsaturated fatty acids, and intracellular pH, hypoxia, and neurotransmitters. Here we attempted to determine whether TREK1 is functionally expressed in bladder cancer 253J cells. K2P channels, including TREK1, TREK2, TASK1, TASK3, and TWIK1, were quantified in cultured human bladder cancer 253J cells using real time quantitative RT-PCR (qRT-PCR) analysis. Among them, TREK1-like channel was recorded at a single channel level using the patch-clamp technique. The TREKl-like channel, with single-channel conductance of ~90 pS at −80 mV, was recorded in symmetrical 150 mM KCl using an excised inside-out patch configuration. The current-voltage relationships were linear and were insensitive to tetraethylammonium. The channel was activated by membrane stretch, free fatty acids, and intracellular acidosis. These results with electrophysiological properties resemble to those of K2P channel, for instance, TREK1. Therefore, we conclude that TREK1 channel is functionally present in bladder cancer 253J cells.

The TREK2 Channel Is Involved in the Proliferation of 253J Cell, a Human Bladder Carcinoma Cell

Park, Kyung-Sun,Han, Min Ho,Jang, Hee Kyung,Kim, Kyung-A,Cha, Eun-Jong,Kim, Wun-Jae,Choi, Yung Hyun,Kim, Yangmi The Korean Society of Pharmacology 2013 The Korean Journal of Physiology & Pharmacology Vol.17 No.6

Bladder cancer is the seventh most common cancer in men that smoke, and the incidence of disease increases with age. The mechanism of occurrence has not yet been established. Potassium channels have been linked with cell proliferation. Some two-pore domain $K^+$ channels (K2P), such as TASK3 and TREK1, have recently been shown to be overexpressed in cancer cells. Here we focused on the relationship between cell growth and the mechanosensitive K2P channel, TREK2, in the human bladder cancer cell line, 253J. We confirmed that TREK2 was expressed in bladder cancer cell lines by Western blot and quantitative real-time PCR. Using the patch-clamp technique, the mechanosensitive TREK2 channel was recorded in the presence of symmetrical 150 mM KCl solutions. In 253J cells, the TREK2 channel was activated by polyunsaturated fatty acids, intracellular acidosis at -60 mV and mechanical stretch at -40 mV or 40 mV. Furthermore, small interfering RNA (siRNA)-mediated TREK2 knockdown resulted in a slight depolarization from $-19.9mV{\pm}0.8$ (n=116) to $-8.5mV{\pm}1.4$ (n=74) and decreased proliferation of 253J cells, compared to negative control siRNA. 253J cells treated with TREK2 siRNA showed a significant increase in the expression of cell cycle boundary proteins p21 and p53 and also a remarkable decrease in protein expression of cyclins D1 and D3. Taken together, the TREK2 channel is present in bladder cancer cell lines and may, at least in part, contribute to cell cycle-dependent growth.

발도르프 유치원 음악교육을 적용한 교회 유치부 교육활동에 대한 고찰

박지연,최양미 안양대학교 2005 人文科學硏究 Vol.13 No.-

The purpose of this study is to search the possibility to apply the music education of Waldorf kindergarten to the church preschool educational activities. The method of this study involved the analyzing of books, articles and related materials about the idea and philosophy of Steiner and Waldorf kindergarten, early childhood music education, Korean church preschool education child. Steiner insisted on the human formation through music education using Pentatonic, music education which emphasized rhythm, music education according to the development course. In Waldorf kindergarten which is based on the educational idea of Steiner, they are proceeding the music education through instruments(Ryer) and 5 note scales, especially while comprising rhythm activity variously such as Rygen, Eurythmie among daily schedule to practice the idea of Steiner who attached importance in rhythm. Now, it is lacking in music education element in church preschool educational activities, so in this viewpoint, if we apply the music education of Waldorf kindergarten to the church preschooler worship and education activities, it will be very effective. Especially, if we apply the integral music education of Waldorf kindergarten that attaches importance to the rhythm in all educational activities, using eurythmie, instruments and 5 note scales, it may be rhythmic worship and activities. But for the music education of Waldorf kindergarten to be newly applied to church preschool education activities, the challenging and researching attitude by religious workers and teachers are necessary, and continuous effort is needed to rightly understand the idea of Steiner and apply it to our situation. Also, the attempt to apply music integrally into every activities, and rhythm activity utilizing eurythmie and education activity utilizing instruments and 5 note scales must be activated, and when these efforts are made continuously, music education may settle newly in Korean churches' preschool group, and the preschoolers will like music through music and will grow up as the preschool children who are familiar with music, and they will grow up to personally matured humans.

접경지역 기업의 연계 및 네트워크의 공간적 특성

박삼옥,이현주,구양미 한국경제지리학회 2004 한국경제지리학회지 Vol.7 No.2

본 연구는 경기도 접경지역에 소재한 기업들을 대상으로 기업연계 및 혁신 네트워크의 공간적 특성을 분석하였다. 이를 통하여 접경지역 기업들의 경제활동의 공간적 특성을 파악하고, 이러한 특성이 기업의 성장 잠재력과 지역경제의 성장에 어떠한 영향을 미쳤는지를 분석하고자 하였다. 분석결과는 다음과 같다. 접경지역 기업의 물자연계의 공간적 특성은 생활권을 중심으로 하는 국지적 연계와 해외지역과의 연계인 광역적 연계가 발달하지 못한 채, 수도권과의 연계만으로 집중되어 있어 기업의 입지가 지역경제 성장에 미치는 효과가 제한적으로 나타나고 있다. 이에 반해 고용연계는 국지적 연계가 상대적으로 높게 나타나고 있으나, 종사자 직급별 거주지 분화 현상이 뚜렷하여 고소득자의 소비 공간이 서울과 신도시로 형성되어 있어 지여사회로의 경제적 파급효과가 미약하게 나타나고 있다. 현재 경기도 접경지역의 중요한 산업공간정책인 산업단지를 분석한 결과 역시 개별입지와 유사한 기업연계의 공간적 특성을 보여주고 있으나, 산업단지에 입지한 기업들이 개별입지 기업보다는 지역의 성장잠재력에 기여하는 것으로 나타났다. 1990년 이후 경기도 접경지역으로의 기업의 입지 증가가 접경지역 경제의 성장잠재력을 높이기 위해서는 국지적, 비국지적 연계가 조화를 이룰수 있는 정책이 요구된다. The purpose of this study is to analyze the spatial characteristics of inter-firm linkages and innovation networks of firms which are located in Gyeonggi border regions. Through this analysis, this study aims at understanding the spatial characteristics of economic activities of firms in Gyeonggi border regions. More importantly, this study focuses on revealing how the spatial characteristics of economic activities of firms affect future growth potentials of firms and regional economic growth. The main findings of this study are as follows. According to the analysis of spatial material linkages, the firms in both footloose location and industrial complex are all proven to be strongly dependent on the Capital Region. Owing to the weak level of the local material linkages, the effects on regional economic growth are limited. But firms in industrial complex contribute more to regional economic growth. The analysis of spatial employment linkages shows that the local linkages are comparatively strong. But the differentiation of residences weaken the positive effects on regional economy. As a concluding remark, the harmony with local linkages and non-local linkages is required to promote future growth potentials of firms and regional economic growth.

Phosphorylation of Focal Adhesion Kinase at Tyrosine 861 Is Crucial for Ras Transformation of Fibroblasts

Lim, Yangmi,Han, Innoc,Jeon, Jihyun,Park, Haein,Bahk, Young-Yil,Oh, Eok-Soo American Society for Biochemistry and Molecular Bi 2004 The Journal of biological chemistry Vol.279 No.28

한국인 Marfan 증후군의 유전자 변이 분석

신미승,박현영,임양미,신길자,장양수,장병철,정남식 대한순환기학회 2003 Korean Circulation Journal Vol.33 No.11

자세 인식 딥러닝을 이용한 교량 자살 방지 시스템

박예빈(Yebin Park),최다선(Dasun Choi),이세인(Sein Lee),정다현(Dahyun Jung),임양미(Yangmi Lim) 한국방송·미디어공학회 2021 한국방송공학회 학술발표대회 논문집 Vol.2021 No.11

Diversity of Ion Channels in Human Bone Marrow Mesenchymal Stem Cells from Amyotrophic Lateral Sclerosis Patients

Kyoung Sun Park,Mi Ran Choi,Kyoung Hwa Jung,SeungHyun Kim,Hyun Young Kim,Kyung Suk Kim,Eun-Jong Cha,Yangmi Kim,Young Gyu Chai 대한생리학회-대한약리학회 2008 The Korean Journal of Physiology & Pharmacology Vol.12 No.6

Human bone marrow mesenchymal stem cells (hBM-MSCs) represent a potentially valuable cell type for clinical therapeutic applications. The present study was designed to evaluate the effect of long-term culturing (up to 10^th passages) of hBM-MSCs from eight individual amyotrophic lateral sclerosis (ALS) patients, focusing on functional ion channels. All hBM-MSCs contain several MSCs markers with no significant differences, whereas the distribution of functional ion channels was shown to be different between cells. Four types of K⁺ currents, including noise-like Ca⁺²-activated K⁺ current (IK_Ca), a transient outward K⁺ current (I_to), a delayed rectifier K⁺ current (IK_DR), and an inward-rectifier K⁺ current (K_ir) were heterogeneously present in these cells, and a TTX-sensitive Na⁺ current (I_Na,TTX) was also recorded. In the RT-PCR analysis, <i>Kv1.1, heag1, Kv4.2, Kir2.1, MaxiK,</i> and <i>hNE-Na</i> were detected. In particular, I_Na,TTX showed a significant passage-dependent increase. This is the first report showing that functional ion channel profiling depend on the cellular passage of hBM-MSCs

The Inhibition of TREK2 Channel by an Oxidizing Agent, 5,5 -dithio- bis (2-nitrobenzoic acid), via Interaction with the C-terminus Distal to the 353rd Amino Acid

Kyoung Sun Park,Hyoweon Bang,Eun-Young Shin,Chan Hyung Kim,Yangmi Kim 대한생리학회-대한약리학회 2008 The Korean Journal of Physiology & Pharmacology Vol.12 No.4

TREK (TWIK-RElated K⁺ channels) and TRAAK (TWIK-Related Arachidonic acid Activated K⁺ channels) were expressed in COS-7 cells, and the channel activities were recorded from inside-out membrane patches using holding potential of −40 mV in symmetrical 150 mM K⁺ solution. Intra</SUP>cellular application of an oxidizing agent, 5,5 -dithio-bis (2-nitrobenzoic acid) (DTNB), markedly decreased the activity of the TREK2, and the activity was partially reversed by the reducing agent, dithiothreitol (DTT). In order to examine the possibility that the target sites for the oxidizing agents might be located in the C-terminus of TREK2, two chimeras were constructed: TREK2 (1-383)/TASK3C and TREK2 (1-353)/TASK3C. The channel activity in the TREK2 (1-383)/TASK3C chimera was still inhibited by DTNB, but not in the TREK2 (1-353)/TASK3C chimera. These results indicate that TREK2 is inhibited by oxidation, and that the target site for oxidation is located between the amino acid residues 353 and 383 in the C-terminus of the TREK2 protein.

Riluzole-Induced Activation of BKCa Channels and Inhibition of Voltage-Gated Na<SUP>+</SUP> Channel in Human Umbilical Cord Vein-Derived Mesenchymal Cells

Kyoung Sun Park,Kyung Hwa Jung,Young Gyu Chai,Chan Hyung Kim,Eun-Young Shin,Yangmi Kim 한국실험동물학회 2008 Laboratory Animal Research Vol.24 No.3

Recently, we demonstrated that large conductance Ca²⁺-activated K⁺ channel (BKCa) and voltage activated Na⁺+ channel (INa) present in human umbilical cord vein-derived mesenchymal cells (hUC-MSCs). However, little is known of single channel characteristic of BKCa in hUC-MSCs. The pharmacological action of riluzole, a drug used for the treatment of amyotrophic lateral sclerosis, has not yet been investigated in stem cells. In this study, we therefore studied effects of riluzole on functional ion channels, BKca and INa, in hUC-MSCs using single channel and/or whole cell configuration. Single channel recording revealed 244.45±29.25 pS of Ca²⁺ and voltage-gated K⁺ channel present in hUC-MSCs and the value is similar to that of typical BKCa channel reported in other cells. Riluzole (100 µM) increased BKCa in cell-attached/whole cell patch configuration. The activation effect of riluzole on this ion channel also found in insideout configuration, suggesting that riluzole may directly activate BKCa in hUC-MSCs. On the contrary, riluzole inhibited INa in a concentration-dependent manner with an IC₅₀ value of 16 µM. Taken together, BKCa and INa in hUC-MSCs can be an important target for the action of riluzole and the channels are partly may contribute to maintaining homeostasis by regulation of ion flux in hUC-MSCs.