偵測微型核糖核酸 miR-524-5p表現量利用原位雜交染色法來作為輔助診斷惡性黑色素瘤的生物標記之研究;Develop the miR-524-5p by in situ hybridization as an ancillary biomarker in melanoma diagnosis

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Title:	偵測微型核糖核酸 miR-524-5p表現量利用原位雜交染色法來作為輔助診斷惡性黑色素瘤的生物標記之研究;Develop the miR-524-5p by in situ hybridization as an ancillary biomarker in melanoma diagnosis
Authors:	王松年;Wang,Sung-Nine
Contributors:	系統生物與生物資訊研究所
Keywords:	微型核糖核酸;miR-524-5p
Date:	2016-07-27
Issue Date:	2016-10-13 13:24:51 (UTC+8)
Publisher:	國立中央大學
Abstract:	本實驗室先前的研究證明，過度表達的miR-524-5p在黑色素瘤中具有抑制細胞增殖、遷移和小鼠腫瘤生長的能力，此外miR-524-5p在黑色素瘤細胞中透過直接抑制BRAF以及ERK2，來抑制MAPK /ERK的訊號。並且在黑色素瘤細胞中， miR-524-5p的表達與MAPK/ ERK訊息傳遞途徑的活性，呈現反比的關聯性。但是目前miR-524-5p在病人中的表達程度是未知的。因此我們的研究目的，在調查人體組織中miR-524-5p的分布狀況是否可以準確地辨別惡性黑色素瘤組織以及良性痣組織，這些結果可以協助確定黑色素瘤病理學在惡性黑色素瘤診斷上的輔助生物標記。我們利用台灣醫院的惡性黑色素瘤組織檢體以及良性痣的組織檢體，一共有58位病人，以及商業化的黑色素瘤組織陣列，總共含有99個黑色素瘤組織和良性痣組織，來研究miR-524-5p和MAPK/ERK訊息傳遞途徑的活性。藉由原位雜交法 (In Situ Hybridization, ISH) 分析miR-524-5p的表達，和免疫組織化學染色 (Immunohistochemistry, IHC) 來偵測MAPK/ ERK訊息傳遞途徑的活性 (透過評估phospho-MEK) 和BRAF的表達。我們的研究結果顯示，這些結果可以協助病理上對惡性黑色素瘤的認定，或更進一步應用於生物標記物用來診斷惡性黑色素瘤組織。;Our previous study demonstrated that over-expression of miR-524-5p suppresses cells proliferation, migration and tumor growth of melanoma in mice. In addition, miR-524-5p plays an important function in regulating MAPK/ERK signaling through directly suppressing BRAF and ERK2 in melanoma cells. In previous study, the expression of miR-524-5p is inversely associated with the activity of the MAPK/ERK pathway in melanoma cells. However, the expression level of miR-524-5p in melanoma patients is unknown and if there are similar inversely expression pattern between MAPK pathway activity and miR-524-5p in human tissue. We investigated the expression of miR-524-5p and MAPK activity using total 58 tissues from melanoma and nevus from Taiwan patients and total 99 melanoma and nevus tissues from commercial tissue array. The expression of miR-524-5p was detected by in situ hybridization (ISH) and the activity of MAPK/ERK pathway (assessed by phosph-MEK) and BRAF expression were measured by immunohistochemistry (IHC). These results could assist to determine the pathology for melanoma or further applied in the biomarker of melanoma diagnosis.
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