目的:关于游离脂肪酸受体2(FFAR2)的蛋白质表达的信息很少,尤其是在肿瘤中。因此,本研究的目的是使用免疫组织化学方法全面表征FFAR2在大量人类正常组织和肿瘤组织中的表达谱,从而为进一步深入研究其潜在的诊断或治疗重要性提供基础.
方法:我们开发了一种新型兔多克隆抗FFAR2抗体,0524,针对人FFAR2的C末端区域。使用表达FFAR2的细胞系BON-1和FFAR2特异性小干扰RNA以及天然和FFAR2转染的HEK-293细胞,通过蛋白质印迹分析和免疫细胞化学证实抗体特异性。然后将抗体用于各种福尔马林固定的免疫组织化学分析,正常和肿瘤人体组织的石蜡包埋标本。
结果:在正常组织中,FFAR2主要存在于大脑皮层的不同细胞群体中,甲状腺的滤泡细胞和C细胞,心脏的心肌细胞,支气管上皮和腺体,肝细胞和肝脏的胆管上皮,胆囊上皮,内分泌胰腺的外分泌和β细胞,肾小球系膜细胞和足细胞以及肾脏的集合管,肠粘膜(特别是肠内分泌细胞),前列腺上皮,睾丸的精细管,和胎盘合胞体滋养层。在肿瘤组织中,FFAR2在甲状腺乳头状癌中特别普遍,甲状旁腺腺瘤,胃,结肠,胰腺,肝细胞,胆管细胞,膀胱,乳房,子宫颈,和卵巢癌。
结论:我们产生并表征了一种新的兔多克隆抗人FFAR2抗体,该抗体非常适合于观察人常规病理组织中的FFAR2表达。该抗体也适用于Western印迹和免疫细胞化学实验。据我们所知,该抗体在各种正常和肿瘤性人体组织中实现了第一个广泛的FFAR2蛋白表达谱.
OBJECTIVE: Little information is available concerning protein expression of the free fatty acid receptor 2 (FFAR2), especially in
tumours. Therefore, the aim of the present study was to comprehensively characterise the expression profile of FFAR2 in a large series of human normal and neoplastic tissues using immunohistochemistry thus providing a basis for further in-depth investigations into its potential diagnostic or therapeutic importance.
METHODS: We developed a novel rabbit polyclonal anti-FFAR2 antibody, 0524, directed against the C-terminal region of human FFAR2. Antibody specificity was confirmed via Western blot analyses and immunocytochemistry using the FFAR2-expressing cell line BON-1 and FFAR2-specific small interfering RNA as well as native and FFAR2-transfected HEK-293 cells. The antibody was then used for immunohistochemical analyses of various formalin-fixed, paraffin-embedded specimens of normal and neoplastic human tissues.
RESULTS: In normal tissues, FFAR2 was mainly present in distinct cell populations of the cerebral cortex, follicular cells and C cells of the thyroid, cardiomyocytes of the heart, bronchial epithelia and glands, hepatocytes and bile duct epithelia of the liver, gall bladder epithelium, exocrine and β-cells of the endocrine pancreas, glomerular mesangial cells and podocytes as well as collecting ducts of the kidney, intestinal mucosa (particularly enteroendocrine cells), prostate epithelium, seminiferous tubules of the testicles, and placental syncytiotrophoblasts. In neoplastic tissues, FFAR2 was particularly prevalent in papillary thyroid carcinomas, parathyroid adenomas, and gastric, colon, pancreatic, hepatocellular, cholangiocellular, urinary bladder, breast, cervical, and ovarian carcinomas.
CONCLUSIONS: We generated and characterised a novel rabbit polyclonal anti-human FFAR2 antibody that is well-suited for visualising FFAR2 expression in human routine pathology tissues. This antibody is also suitable for Western blot and immunocytochemistry experiments. To our knowledge, this antibody enabled the first broad FFAR2 protein expression profile in various normal and neoplastic human tissues.