• Asian Pacific Journal of Cancer Prevention
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• 제14권2호
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• pp.639-643
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• 2013

Aim: Connexin 43 (Cx43) and E-cadherin are important biomarkers related with cancer. Their expression at protein and mRNA levels was here investigated in 50 primary lung carcinoma tissues and 20 samples of adjacent normal tissue of Chinese patients with non-small cell lung cancer (NSCLC). Methods: Protein and mRNA expression were evaluated by ABC immunohistochemistry and RT-PCR. Results: (1) The positive expression rates of Cx43 and E-cadherin protein were higher in the adjacent normal tissues than those in the primary lung carcinoma tissues; (2) the positive expression rates of Cx43 and E-cadherin protein decreased with NSCLC progression; (3) the expression of E-cadherin protein was not related with the pathological type of NSCLC; and (4) the relative quantity of the Cx43 or E-cadherin mRNA expression was correlated with the the histological type, clinical stage, cancer cell differentiation and the lymph node metastasis. Conclusion: The data suggested that the Cx43 and E-cadherin are reduced with NSCLC progression, and might be important biomarkers for judging the metastasis and prognosis.

• 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.

• 한국발생생물학회지:발생과생식
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• 제18권4호
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• pp.293-300
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• 2014

Direct cell-cell communication through connexin (Cx) complexes is a way to achieve functional accordance of cells within a tissue or an organ. The initial segment (IS), a part of the epididymis, plays important roles in sperm maturation. Steroid hormones influence on expression of a number of genes in the IS of adult animals. However, developmental effect of sex hormones on the gene expression in the IS has not been examined. In this study, estradiol benzoate (EB, an estrogen agonist) or flutamide (Flu, an androgen antagonist) was exogenously administrated at 1 week of postnatal age, and expressional changes of Cx genes in the IS were determined at 4 months of age by a quantitative real-time PCR analysis. Treatment of EB at $0.015{\mu}g/kg$ body weight (BW) increased expression of Cx30.3, 31.1, and 43 genes. However, treatment of 1.5 mg EB/kg BW resulted in expressional decreases of Cx31, 32, and 45 genes and caused increases of Cx30.3 and 43 gene expression. Significant decreases of Cx31, 31.1, 32, 37, and 45 gene expression were detected with a treatment of $500{\mu}g\;Flu/kg$ BW, while expression of Cx43 gene was significantly increased with a treatment of $500{\mu}g\;Flu/kg$ BW. A treatment of $50{\mu}g\;Flu/kg$ BW led to significant increases of Cx30.3, 32, 37, 40, and 43 gene expression. These findings imply that exogenous exposure of steroidal hormones during the early developmental period would result in aberrant expression of Cx genes in the adult IS.

• Reproductive and Developmental Biology
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• 제34권2호
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• pp.103-109
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• 2010

Functional regulation of a specific tissue or organ is controlled by a number of ways, including local cell-cell interaction. Of several forms of cell-cell junctional complexes, gap junctions are caught a great attention due to a formation of direct linkage between neighboring cells. Gap junctions are consisted of connexin (Cx) isoforms. In the present study, we evaluated expressional profiling of Cx isoforms in the rat initial segment (IS) of the male reproductive tract at different postnatal ages. The presence and expression of 13 Cx isoform mRNAs were determined by semi-quantitative real-time PCR analyses. A total of 8 Cx isoform mRNAs were detected in the IS of the male rats during postnatal development. The highest level of Cx30.3 mRNA was found at 5 months of age, while abundance of Cx31 mRNA was the highest at 1 year of age. Expression of Cx31.1 gene was relatively consistent during the postnatal development. Fluctuation of Cx32 and 37 gene expression was observed during the postnatal period. Significant elevation of Cx40 mRNA abundance was detected at 25 days of age and older ages. Expression patterns of Cx43 and 45 genes were similar with the highest level at 2 weeks of age, followed by gradual decreases at older ages. These results indicate differential regulation on expression of Cx isoforms in the rat IS during postnatal development. A complicated regulation of gene expression of Cx isoforms in the IS at different postnatal ages is suggested.

• 한국발생생물학회지:발생과생식
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• 제22권1호
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• pp.29-38
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• 2018

In the multicellular tissue, cell-cell interaction is important for a precise control of its function. The exchange of signaling molecules between adjacent cells via connexon allows the functional harmony of cells in the tissue. The present research was to determine the presence and expressional patterns of connexin (Cx) isoforms in the rat epididymal fat during postnatal development using quantitative real-time polymerase chain reaction (PCR) analysis. Of 13 Cx isoforms examined, expression of 11 Cx isoforms in the epididymal fat during postnatal development was detected. These Cx isoforms include Cx26, Cx31, Cx31.1, Cx32, Cx33, Cx36, Cx37, Cx40, Cx43, Cx45, and Cx50. Expressional levels of all Cx isoforms at 1 and 2 years of age were significantly higher than those at the early postnatal ages, such as 7 days, 14 days, and 24 days of ages. Except Cx33 and Cx43, the transcript levels of rest Cx isoforms at 1 year of age were significantly lower than that at 2 years of age. In addition, expressional patterns of Cx isoforms between 7 days and 5 months of ages generally varied according to the isoform. The existence of various Cx isoforms in the rat epididymal fat has been identified and expression of each Cx isoform in the epididymal fat during postnatal development has shown a particular pattern, distinguishable from the others. To our knowledges, this is the first report showing expressional patterns of Cx isoforms at transcript level in the epididymal fat at various postnatal ages.

• Biomolecules & Therapeutics
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• 제11권4호
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• pp.224-231
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• 2003

The aim of this study was to investigate the effect of resveratrol on the oxidative stress-induced inhibition of gap junctional intercellular communication in HaCaT keratinocytes. Anti-oxidative activity of resveratrol was measured by $\alpha,\alpha$-diphenyl-$\beta$-picrylhydrazyl assay and dichlorodihydrofluorescein diacetate oxidation assay. Gap junctional intercellular communication in HaCaT keratinocytes was assessed using the scrape loading/dye transfer technique. Western blots and reverse transcription-polymerase chain reaction were also analyzed for connexin 43 protein and mRNA expression, respectively. Resveratrol scavenged directly the stable $\alpha,\alpha$-diphenyl-$\beta$-picrylhydrazyl radical over a concentration range of 4 mg/ml ($78.2{\pm}2.7$% of control) to 500 mg/ml ($29.9{\pm}4.2$% of control) and decreased the intracellular reactive oxygen species induced by ultraviolet A (UVA) irradiation ($89.3{\pm}1.1$% of UVA group), ultraviolet B (UVB) irradiation ($70.9{\pm}1.7$% of UVB group) and 12-0-tet-radecanoylphorbol-13-acetate (TPA, $48.3{\pm}1.1$% of TPA group), respectively. UVA irradiation and TPA markedly reduced gap junctional intercellular communication, which was restored by resveratrol. There were no significant differences in the level of connexin 43 protein and mRNA expression among any of the experimental groups. Our data suggests that resveratrol has the protective effect on the oxidative stress-induced inhibition of gap junctional intercellular communication in HaCaT keratinocytes, and this protection is likely due to the scavenging of reactive oxygen species.

• Clinical and Experimental Pediatrics
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• 제45권3호
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• pp.354-361
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• 2002

목 적: 본 연구에서는 in vitro 및 in vivo에서 HSV-TK 유전자 치료의 bystander 효과 및 기전을 관찰하고, 싸이토카인 유전자의 병합 치료가 bystander 효과를 증폭시킬 수 있는지 조사하여 그 결과를 향후 신경모세포종 치료에 적용하고자 본 연구를 시행하였다. 방 법: A/J 마우스 신경모세포종 모델에서, neuro-2a/TK 세포와 unmodified neuro-2a세포를 여러 비율로 혼합하여 접종한 후 GCV를 투여하여 종양의 크기 변화 및 생존 유무를 관찰하였다. 마우스 신경모 세포종에서 bystander 효과의 기전을 알아보기 위해 neuro-2a/TK 세포를 주입한 마우스의 조직을 절개하여 connexin 43, CD4+ 및 CD8+ 세포 침윤을 관찰 하였다. 10% 및 25% HSV-TK 투여 군에서 neuro-2a/IL-2 세포의 투여가 bystander 효과를 증폭시키는 지 조사하였다. 결 과 : 1) in vitro 및 in vivo에서 bystander 효과는 뚜렷하게 관찰되었다. 2) 마우스 신경모세포종의 면역 조직 화학 염색 검사에서 connexin 43 발현은 관찰되지 않았으나 많은 수의 CD4+ 및 CD8+ T 세포의 침윤이 관찰되었다. 3) 10% 및 25% HSV-TK 투여군의 마우스에서 neuro-2a/IL-2 세포의 투여는 대조군과 비교하여 종양의 성장을 더 억제 시켰다. 결 론: In vitro 및 in vivo에서 HSV-TK/GCV 유전자 치료의 bystander 효과는 뚜렷하였으며, 그 기전으로 면역 세포가 중요한 역할을 할 것으로 추측된다. 그리고 bystander 효과는 IL-2 투여에 의해 증폭됨을 확인하였다. 이는 향후 신경모세포종의 수술 후 잔존암 치료에 HSV-TK/GCV 유전자 치료가 이용 가능할 것으로 생각된다.

• 한국임상수의학회:학술대회논문집
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• 한국임상수의학회 2006년도 추계학술대회 및 정기총회
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• pp.57-57
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• 2006

• The Korean Journal of Physiology and Pharmacology
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• 제6권4호
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• pp.173-181
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• 2002

The sinoatrial (SA) node is a complex and inhomogeneous tissue in terms of cell morphology and electrical activity. There are two models of the cellular organisation of the sinoatrial node: the gradient and mosaic models. According to the gradient model there is a gradual transition in morphology and electrical properties of SA node cells from the centre to the periphery of the SA node. In the mosaic model, there is a variable mix of atrial and sinoatrial node cells from the centre to the periphery. This review focuses on the cellular organisation of the rabbit sinoatrial node in terms of the expression of connexin (Cx40, Cx43 and Cx45), L-type $Ca^{2+}$ channel and $Na^+-Ca^{2+}$ exchanger proteins. These immunocytochemical data, together with morphological and electrophysiological data, obtained from the intact sinoatrial node and isolated sinoatrial node cells support the gradient model of the cellular organisation of the SA node. The complex organisation of the sinoatrial node is important for the normal functioning of the sinoatrial node: (i) it allows the sinoatrial node to drive the surrounding hyperpolarized atrial muscle without being suppressed by it; (ii) it helps the pacemaker activity of the sinoatrial node continue under a wide range of physiological and pathophysiological conditions; (iii) it helps protect the sinoatrial node from reentrant arrhythmias.

• 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.