사람 상피세포주(HaCaT)에서의 간극결합 세포 간 신호전달의 조절

강희선; 김병철; 강성욱; 홍정주; 전형진; 정연훈

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자료유형: 학술저널

저자정보: 강희선 김병철 강성욱 홍정주 전형진 정연훈

저널정보: 대한이비인후과학회 대한이비인후-두경부외과학회지 대한이비인후과학회지 두경부외과학 제53권 제11호

발행연도: 2010.1

수록면: 675 - 685 (11page)

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연구주제

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연구배경

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연구방법

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Background and Objectives Unlike the normal skin, cholesteatomas characterized by hyperproliferative keratinocytes exhibits up-regulation of connexins (Cxs) and gap junctional intercellular communication (GJIC). Currently, there are no appropriate clinical methods that can inhibit cholesteatoma progression nor are there available optimal in vitro models of cholesteatomas. The objectives of this study were to identify the regulating materials that control GJIC using human keratinocyte cells (HaCaT) and to get preliminary information about how to inhibit cholesteatoma progression with an aim to make in vitro models. Materials and Method Acetic acid (AA), H2O2, dexamethasone, retinoic acid (RA), or green tea extracts-epicatechin (EC) and epigallocatechin gallate (EGCG) were used for this study. After HaCaT cells were cultured with chemicals for 24 hours, cytotoxicity was quantitatively analyzed by cell counting and Neutral-red uptake test. Reverse transcriptase-polymerase chain reaction, Western blot and immunocytochemistry were performed to analyze the change of Cx expression. GJIC was functionally evaluated with scrape-loading dye transfer (SLDT). Results After the 24-hour culture, H2O2 or EGCG (100 μM) were observed to have interfered with cell growth. In the Western blot, Cx26 and Cx30 showed higher up-regulation by EGCG or dexamethasone, but less down-regulation by AA or H2O2 than the control. In comparison with the control, immunocytochemistry (Cx26, Cx43) showed less expression and abnormal location of Cxs under AA, H2O2, or 50 μM EGCG than the control, and increased up-regulation or equal expression under 5μM EGCG, EC, RA, or dexamethasone was greater than the control. In SLDT, dye transfer was significantly lower in AA-, H2O2-, dexamethasone-, or RA-treated cells than in the control cells. EC showed higher dye transfer than the control cells. Conclusion The expression of Cxs and GJIC on human HaCaT keratinocytes can be up- or down-regulated by chemicals such as AA, H2O2, dexamethasone, or EC. These results may be useful information in understanding the progression or inhibition mechanisms of cholesteatomas. Korean J Otorhinolaryngol-Head Neck Surg 2010;53:675-85

#Gap junctions ㆍConnexins ㆍCholesteatoma ㆍKeratinocytes ㆍIntercellular Key WordsZZcommunication.

참고문헌 (21)

참고문헌 신청

Alexander M. Raynov, / 2008 / Establishment and Characterization of an In Vitro Model for Cholesteatoma / Clinical and Experimental Otorhinolaryngology 1 (2) : 86 ~ 91

Ara C / 2002 / Retinoic acid modulates gap junctional intercellualr communication in hepatocytes and hepatoma cells / Cell Mol Life Sci 59 (10) : 1758 ~ 1765

Bevans CG / 1998 / Isoform composition of connexin channels determines selectivity among second messengers and uncharged molecules / J Biol Chem 273 (5) : 2808 ~ 2816

Butterweck A / 1994 / Differential expression of the gap junction proteins connexin45, -43, -40, -31, and -26 in mouse skin / Eur J Cell Biol 65 (1) : 152 ~ 163

Choung YH / 2006 / Expression of the gap junction proteins connexin 26 and connexin 43 in human middle ear cholesteatoma / Acta Otolaryngol 126 (2) : 138 ~ 143

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