• Asian Pacific Journal of Cancer Prevention
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• 제13권1호
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• pp.205-209
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• 2012

Background: The inhibition of tumor cell growth without toxicity to normal cells is an important target in cancer therapy. One possible way to increase the efficacy of anticancer drugs and to decrease toxicity or side effects is to develop traditional natural products, especially from medicinal plants. Paris polyphylla Smith has shown anti-tumour effects by inhibition of tumor promotion and inducement of tumor cell apoptosis, but mechanisms are still not well understood. The present study was to explore the effect of Paris polyphylla Smith extract (PPSE) on connexin26 and growth control in human esophageal cancer ECA109 cells. Methods: The effects of PPSE on Connexin26 were examined by RT-PCR, western blot and immunofluorescence; cell growth and proliferation were examined by the 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyl-tetrazolium bromide (MTT) assay. Results: PPSE inhibited the growth and proliferation on esophageal cancer ECA109 cells, while increasing the expression of connexin26 mRNA and protein; conversely, PPSE decreased Bcl-2 and increased Bad. Conclusion: This study firstly shows that PPSE can increase connexin26 expression at mRNA and protein level, exerting anti-tumour effects on esophageal cacner ECA109 cells via inhibiting cell proliferation and inducing cell apoptosis.

• Journal of Animal Science and Technology
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• 제49권1호
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• pp.15-24
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• 2007

본 연구의 목적은 생후 발달과정에서 숫 백서 생식기의 일부인 efferent ductules (ED)에서 다양한 connexin 이형질체의 발현과 양상을 알아보았다. 1주, 2주, 1개월, 3개월 그리고 6개월령의 백서의 ED로부터 total RNA을 분리하였으며, semi-quantitative RT-PCR 방법을 사용하여 총 14개 connexin 이형질체의 mRNA 발현 양상을 알아보았다. 백서의 ED에서 연구된 14개 중 6개 connexin 이형질체의 발현이 보여졌으며, connexin 26과 30의 mRNA 발현은 연령에 따라 심한 변화를 보여주었다. Connexin 31.1의 발현은 1달령의 ED에서 가장 낮게 나타났으며, connexin 37과 45는 주로 초기 발달 과정에서 높은 수준으로 발현됨을 보여주었다. 또한, connexin 43의 발현은 1주령에서 가장 낮았으며 2주령 이후에는 큰 변화없이 일정한 수준을 유지하였다. 본 연구는 백서의 ED에서 다양한 종류의 connexin 이형질체가 발현되며, 이러한 발현은 생후 연령에 따라 서로 다르게 조절됨을 보여준다.

• 대한수의학회지
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• 제45권4호
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• pp.517-525
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• 2005

This study was performed to evaluate using wild-type (WT) and $C{\times}32$ knockout (KO) mice if lack of cell to cell communication by connexin 32 gap junction enhances the benzene-induced hematotoxicity and hemopoietic tumor development. The WT and $C{\times}32$ KO mice were exposed to 300 ppm of benzene for 6 hours/day, 5 days/week for a total of 26 weeks by inhalation, and then sacrificed to evaluate the toxicities of hemopoietic organs or allowed to live out their life span to evaluate the hemopoietic tumor incidence. The significant increase and decrease of organ weight were respectively noted in spleen and thymus of both WT and $C{\times}32$ KO mice without significant difference between the genotypes. Histopathologically, benzene exposure for 26 weeks induced the morphological changes in hemopoietic organs, characterized by fat cell accumulation in the bone marrow and extramedullary hemopoiesis in the spleen. The fat cell accumulation was, compared with that of WT mice, considerably exacerbated in the $C{\times}32$ KO mice. However, no significant difference was observed in the changes of hematological values and bone marrow cellularity as well as in the onset and incidence of hemopoietic tumors between WT and $C{\times}32$ KO mice. In conclusion, this study indicated little significant role of the cellular communication by $C{\times}32$ gap junction in the action mechanism of benzene hematotoxicity and leukemogenicity.

• 생명과학회지
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• 제17권11호
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• pp.1517-1522
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• 2007

간극결합(intercellular channel)은 인접하는 두 세포사이에 형성된 이온채널이며 이를 통해서 각종 이온, 이차 신호전달물질, 그리고 1 kDa 미만의 대사물질들이 통과한다. 아울러, sodium 혹은 potassium 이온채널처럼 반쪽의 간극결합(connexon 혹은 hemichannel)도 세포막채널로서 작용을 한다. 현재까지 간극결합을 구성하는 connexin (Cx) 단위체는 26종류 이상이 확인되었다. 이 가운데, Cx32, Cx38, Cx46 그리고 Cx50 만이 간극결합채널뿐만 아니라 세포막채널로서도 기능을 수행한다. Xenopus oocytes에서 connexin 38 (Cx38)이 발현하는 것으로 알려져 있지만 Cx38의 생물리학적 특성이 단일채널수준에서 연구가 진행된 경우는 없다. 이번 연구에서는 Cx38 채널의 생물리학적 특성, 즉 전압-의존적 개폐와 투과성(전기전도도와 이온선택성)을 알아보고자 단일채널기록을 수행하였다. Cx38 hemichannel은 전압-의존적인 빠른 개폐와 느린 개폐의 특성을 보였다. 양성전압 환경에서는 Cx38 채널이 낮은 열릴 확률(open probability)로 빠른 개폐가 유도된 반면, 음성전압에서는 느린 개폐가 높은 열릴 확률로 유도되었다. bi-ionic 실험을 통하여, Cx38 채널은 양이온보다 음이온을 더 선택 적으로 통과시킨다는 점을 알게 되었다. Cx38의 아미노산서열을 살펴보면, 아미노말단부위에 전하를 띠는 5개의 아미노산 잔기가 존재한다. 앞으로 이들 잔기를 치환시킨 돌연변이 Cx38 채널을 이용하여 과연 이들 아미노산 부위가 전압-의존적 개폐와 투과성에 관여하는 지 여부를 조사하는 연구는 매우 흥미로운 결과를 도출할 것으로 기대한다.

• 대한수의학회지
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• 제43권4호
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• pp.649-656
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• 2003

We have a cultured method to grow rat mammary epithelial cells (RMEC) for 1 to 14 days in 1:1 mixture of Dulbecco's Modified Eagle Medium: Nutrient and F-12 (DMEM/F-12) containing 10% fetal bovine serum (FBS), human EGF, insulin, hydrocortisone, human transferrin and $17{\beta}$-estradiol in vitro. We were able to isolate and distinguish two cell types, luminal epithelial cells and myoepithelial cells, from primary clutures of RMEC. Immunocytochemical stains were used to distingusih luminal epithelial cells and myoepithelial cells. Peanut lectin (PNA) was stained in most alveolar epithelail cells and luminal epithelial cells of rats, while Thy-1.1, a maker of potential rat mammary myoepithelial cells, was expressed in myoepithelial cells in the rat. Also, we examined the expression patterns of three types of gap junction proteins, connexin 26 ($C{\times}26$), connexins 32 ($C{\times}32$) and connexin 43 ($C{\times}43$) by immunocytochemistry and western blot analysis. In the cell types, the results show that at the early stage of culture, luminal epithelial cells were increased and these cells were surrounded by myoepithelial cells. At the late stage of culture, luminal epithelial cells were decreased, in contrast myoepithelial cells were increased. In the expression pattern of gap junction, $C{\times}26$ maintained it's expression until day 3, but afterwards gradually decreased in intensity. Expression of $C{\times}32$ remained until day 5, then decreased slightly. $C{\times}43$ gradually increased untill the middle time of culture then decreased in intensity. These results suggest that connexins may be important for the control of growth in rat mammary epithelial cell types.

• 대한치과교정학회지
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• 제31권5호
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• pp.525-534
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• 2001

치아이동시 골세포간 활성 조차에서 세포간 교통의 중요성이 차츰 인식되고 있으며 세포간 교통을 위한 교통반의 존재가 확인되고 있다. 동물실험 모델에서 실험적 치아이동을 통해 조성된 압박 및 견인측 치주인대에서 교통반 단백의 일종인 connexin 43의 발현을 관찰함으로써 인접 연조직 및 골 조직 내에서 세포 신호 전달 양상의 한 부분을 파악하고자 하였다. Sprague-Dawley계 백서 27마리를 대조군(3마리)과 실험군(24마리)으로 나누었으며, 실험군은 견인력(75g)을 가한 후 12시간, 1일, 4일, 7일, 14일, 28일이 경과한 후 각각 4마리씩 희생시켜, connexin 43의 발현을 면역조직화학적으로 관찰한 바 다음과 같은 결과를 얻었다. 1. 대조군에서의 connexin 43 발현은 치은, 상아질, 치주인대, 치수, 골세포에서 경미하였다. 2. 교정적 치아이동에 반응하는 치수, 치주인대, 골모세포, 파골세포, 골세포부위에서 connexin 43의 발현이 많았으나 치은, 상아질, 상아모세포에서의 발현은 견인력 적용기간에 관계없이 대조군과 큰 차이가 없었다. 3. 치수에서 connexin 43의 발현은 교정력을 가한 4일에서부터 증가하여 7일째까지 크게 증가한 후 14일째부터 감소되었으며 28일째에는 대조군과 차이가 없었다. 4. 치주인대에서 connexin 43의 발현은 주로 혈관을 중심으로 치조골 면을 따라 압박측에서 주로 나타났는데 교정력을 가한 12시간에서부터 증가하여 4일에서 가장 많은 발현을 보인 후 7일부터 감소하기 시작하였다. 5. 골모세포 및 골세포에서 발현은 실험 1일째부터 출현해 4일째에 가장 많은 발현을 보였고 7일 이후에는 크게 감소하였으며 파골세포에서의 발현은 골모세포와 골세포보다 실험 4일째에 더 많이 관찰할 수 있었다.기계화와 자동화(26.2%), 정성부족으로 인한 맛의 감소(21.6%) 순으로 나타났고, 성별(p<0.01), 연령별(p<0.05)로 유의적 인 차이를 보였다. 7. 전통음식 패스트푸드점이 생기면 이용도에 관한 견해는 '가끔 이용할 것이다'가 56.5%, '많이 이용할 것이다'가 19.8%로 나타났고, 연령별(p<0.05), 주거상태별(p<0.05)로 유의적인 차이를 보였는데 연령이 많을수록 '이용치 않겠다'는 비율이 높았고, 특히 주거상태별로는 '전혀 이용치 않을 것이다'의 비율이 하숙생이 높게 나타났다. 본 연구는 유행에 민감하고 문화의 변화를 주도하며 패스트푸드를 가장 많이 이용하는 세대인 대학생들을 대상으로 우리의 전통음식을 패스트푸드화 하였을 때의 견해를 알아보고 그 전망을 살피기 위한 연구였다. 연구결과에서 나타난 바와 같이 유행의 첨단을 달리는 신세대라 하더라도 많은 비율의 대학생들이 전통음식의 패스트푸드화에 전망을 밝게 보고 있었으며 이를 위해서는 많은 문제점도 지적하였다. 이에 전통음식의 패스트푸드화와 상품화에 대한 인식이 고무적인 방향으로 변화하고 있어 앞으로 계속 그 추이를 연구할 가치가 있다고 사료된다.ematured infants during the early weeks of life. 모든 치근단 수준에서 비표준화 medium 크기 master cone 사용군이 ISO 표준화 규격의 master cone 사용군에 비해 유의하게 높은 gutta-percha 면적비를 나타내었다 (p < 0.01). 비표준화 medium크기 master cone 사용군에서는 표준화 규격의 master cone 사용군에 비해 유의하게 적은 수의 accessory cone이 사용되었다 (p < 0.01). 대학생간에는 유의한 차이(p<0.05)가 인정되었다. 응답자의 체형은

• 한국발생생물학회지:발생과생식
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• 제19권4호
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• pp.217-226
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• 2015

A proper development of the epididymis during the early postnatal development is required for successful fertility in the adult male. Direct cell-cell communication via connexin (Cx) molecules is a common way of cellular interactions to achieve normal development of a given tissue consisting of different cell types. The present research was attempted to determine the effect of exogenous exposure to estrogenic agonist or antiandrogen at the weaning age on expression of Cx isoforms in the adult corpus epididymis. Male rats were subcutaneously administrated with estradiol benzoate (EB) or flutamide (Flu) at the weaning age. The tissue was collected at 4 months of age. Expressional levels of Cx isoforms were determined by a quantitative real-time PCR. Statistical comparison showed significant increases of Cxs31, 32, 37, 40, and 43 transcript amounts by a treatment of $0.015{\mu}g$ of EB /kg body weight (BW). A treatment of $1.5{\mu}g$ of EB /kg BW caused a significant decrease of Cx43 gene expression but increases of Cxs26, 31, 32, 37, and 40 transcript levels. Exposure to $500{\mu}g$ of Flu/kg BW induced an increase of Cx37 expression but significant decreases of Cxs43 and 45 mRNA levels. Expression of Cx37 was increased by a treatment of 5 mg of Flu/kg BW, while transcript levels of Cxs26, 30.3, 31, 31.1, 32, and 43 were significantly decreased by same treatment. These results demonstrate that exposure to steroidal compounds at the early developmental age alters expression of Cx isoforms in the adult corpus epididymis.

• Genomics & Informatics
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• 제8권2호
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• pp.70-75
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• 2010

Asp66his, Asp54Lys, and Asp50Asn are mutations in connexin 26 that are observed in the clinic and give rise to autosomal dominant syndromes. They are the result of point mutations in the human gap junction ${\beta}-2$ gene. In order to investigate the structural mechanism of Bart-Pumphrey Syndrome, Keratitis-Ichthyosis-Deafness Syndrome, and Vohwinkel Syndrome, homology modeling was carried out. Asp66 has direct contact with Asn62 by two hydrogen bonds in the wild-type protein, and in Asp66His, the biggest change observed is a tremendous energy increase caused by hydrogen bond breakage to Asn62. Shifts in the side chain and new hydrogen bond formation are observed for Lys54 compared to the wild-type protein (Asn54) and result in closer contact to Val84. Asp50Asn causes a significant decrease in bond energy, and residual charge reversal repels the ion and metabolites and, hence, inhibits their transportation. Such perturbations are likely to be a factor contributing to abnormal functioning of ion channels, resulting cell death and disease.

• Journal of Microbiology and Biotechnology
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• 제21권5호
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• pp.477-482
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• 2011

There are accumulating evidences suggesting that connexin (Cx), a gap junction channel-forming protein, acts as a growth suppressor in various cancer cells, and this effect is attributeed to the gap junction-mediated intercellular communication (GJIC). In order to characterize the relationship between the growth-arresting activity of Cx26 and its cytoplasmic localizations after expression, we linked a nuclear export signal (NES) sequence to Cx26 cDNA before transfecting into a rat breast cancer cell line. A confocal fluorescent microscopic observation revealed that the insertion of NES minimized the nuclear expression of Cx26, and increased its cytoplasmic expression, including plasma membrane junctions. Total cell counting and BrdUrd-labeling experiments showed that the growth of the breast cancer cells was inhibited by 74% upon transfection of Cx26-NES, whereas only 9% inhibition was observed with only Cx26 cDNA.

• Journal of Animal Science and Technology
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• 제55권4호
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• pp.249-255
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• 2013

Intercellular interactions are important for the proper development and regulation of tissue function. This is especially necessary in the epididymis, a part of male reproductive tract where sperm become mature and acquire their fertilizing capacity. The caput region of the epididymis consists of several types of cells, including principal, basal, and apical cells. Direct intercellular communication is thus required to precisely regulate the functions of the caput epididymis. In this regard, connexin (Cx) is a molecule that forms channels, which allow the direct exchange of small molecules between cells, enabling intercellular communication. In this study, the expression of Cx isoforms in the caput epididymis at different postnatal ages was determined by using quantitative real-time polymerase chain reaction analysis. Nine of 13 Cx isoforms were detected. The transcript levels of Cx30.3, 31, 31.1, 32, and 40 were highest at 45 days of age, while the expression of Cx43 and 45 gradually decreased with age. A substantial fluctuation of Cx26 expression was detected, with significant decreases before and during puberty, followed by a transient increase at adult-hood and rapid decreases at an old age. A significant increase in Cx37 transcript was observed at 25 days of age, followed by gradual decreases at adult and old ages. These results indicate the significant differential expression of various Cx isoforms in the caput epididymis during postnatal development. It further suggests that the functional regulation and developmental maturation of the caput epididymis are highly related to the postnatal age-related differential expression of Cx isoforms.