Greig Aina V H, Linge Claire, Healy Vourneen, Lim Philip, Clayton Elizabeth, Rustin Malcolm H A, McGrouther D Angus, Burnstock Geoffrey
Autonomic Neuroscience Institute, Royal Free and University College Medical School, Royal Free Campus, London, UK.
J Invest Dermatol. 2003 Aug;121(2):315-27. doi: 10.1046/j.1523-1747.2003.12379.x.
We investigated the use of purinergic receptors as a new treatment modality for nonmelanoma skin cancers. Purinergic receptors, which bind adenosine 5'-tri-phosphate, are expressed on human cutaneous keratinocytes. Previous work in rat and human epidermis suggested functional roles for purinergic receptors in the regulation of proliferation, differentiation, and apoptosis. Immunohistochemical analysis of frozen sections in human basal cell carcinomas and squamous cell carcinomas for P2X5, P2X7, P2Y1, P2Y2, and P2Y4 receptors was performed, accompanied by detailed analysis of archive material of tumor subtypes in paraffin sections. Functional studies were performed using a human cutaneous squamous cell carcinoma cell line (A431), where purinergic receptor subtype agonists were applied to cells and changes in cell number were quantified via a colorimetric assay. Immunostaining in paraffin sections was essentially the same as that in frozen sections, although more detail of the subcellular composition was visible. P2X5 and P2Y2 receptors were heavily expressed in basal cell carcinomas and squamous cell carcinomas. P2X7 receptors were expressed in the necrotic center of nodular basal cell carcinomas and in apoptotic cells in superficial multifocal and infiltrative basal cell carcinomas, and squamous cell carcinomas. P2Y1 receptors were only expressed in the stroma surrounding tumors. P2Y4 receptors were found in basal cell carcinomas but not in squamous cell carcinomas. P2X5 receptors appear to be associated with differentiation. The P2X7 receptor agonist benzoylbenzoyl-adenosine 5'-triphosphate and high concentrations of adenosine 5'-triphosphate (1000-5000 microM) caused a significant reduction in A431 cell number (p<0.001), whereas the P2Y2 receptor agonist uridine 5'-triphosphate caused a significant amount of proliferation (p<0.001). We have demonstrated that non-melanoma skin cancers express functional purinergic receptors and that P2X7 receptor agonists significantly reduce cell numbers in vitro.
我们研究了嘌呤能受体作为非黑素瘤皮肤癌新治疗方式的应用。与5'-三磷酸腺苷结合的嘌呤能受体在人皮肤角质形成细胞上表达。先前在大鼠和人表皮中的研究表明嘌呤能受体在增殖、分化和凋亡调节中具有功能作用。我们对人基底细胞癌和鳞状细胞癌的冰冻切片进行了P2X5、P2X7、P2Y1、P2Y2和P2Y4受体的免疫组织化学分析，并对石蜡切片中肿瘤亚型的存档材料进行了详细分析。使用人皮肤鳞状细胞癌细胞系（A431）进行功能研究，将嘌呤能受体亚型激动剂应用于细胞，并通过比色法对细胞数量变化进行定量。石蜡切片中的免疫染色与冰冻切片基本相同，尽管亚细胞组成的细节更清晰可见。P2X5和P2Y2受体在基底细胞癌和鳞状细胞癌中大量表达。P2X7受体在结节性基底细胞癌的坏死中心以及浅表多灶性和浸润性基底细胞癌及鳞状细胞癌的凋亡细胞中表达。P2Y1受体仅在肿瘤周围的基质中表达。P2Y4受体在基底细胞癌中发现，但在鳞状细胞癌中未发现。P2X5受体似乎与分化有关。P2X7受体激动剂苯甲酰苯甲酰-5'-三磷酸腺苷和高浓度的5'-三磷酸腺苷（1000 - 5000 microM）导致A431细胞数量显著减少（p<0.001），而P2Y2受体激动剂5'-三磷酸尿苷则导致大量增殖（p<0.001）。我们已经证明非黑素瘤皮肤癌表达功能性嘌呤能受体，并且P2X7受体激动剂在体外可显著减少细胞数量。
