腺苷对葡萄膜黑色素细胞和葡萄膜黑色素瘤细胞增殖的影响

目的探讨腺苷受体在正常人葡萄膜黑色素细胞和葡萄膜黑色素瘤细胞中的表达以及腺苷对其细胞增殖能力的影响。方法体外培养正常人葡萄膜黑色素细胞（U95）和葡萄膜黑色素瘤细胞（$65），采用Real—timePCR检测4种不同亚型腺苷受体（、A1、A2A、A2B、A3）mRNA在U95和S65中的表达情况。以0、0．1、1、10和100μmol／L的腺苷分别作用U95和S65细胞24h和48h后，采用MTT法检测细胞光密度值和抑制率，分析不同时间、不同浓度的腺苷对U95和S65增殖能力的影响。对上述所得数据进行单因素方差分析或独立样本t检验。结果U95和S65细胞中4种类型腺苷受体A1、A2A、A3的mRNA均有表达。在U95细胞中，A1、A孙A2B、A3受体mRNA表达值分别为1．32±0．47、0．28±0．05、1．03±0．10、0．20±0．07，四者之间差异有统计学意义（F=15．90，P〈0．01），A1和A2B的mRNA表达水平相似，且明显高于A2A和凡的表达；在$65细胞中，A1、A2A、A3气受体mRNA表达值分别为1．03±0．27、2．23±0．35、127．34±18．69、0．17±0．07，四者之间差异有统计学意义（F=163．20，P〈0．01），A2HmRNA表达水平明显高于其他3种腺苷受体亚型。A2A和A。在S65中的表达明显高于其在U95中的表达量（A2A：t=9．35，P〈0．0l；A2B2Bt=11．43，P〈0．01）。作用时间24h，0．1、1、10和100μmol／L腺苷对S65细胞的抑制率分别为3．47％、14．93％、19．79％和26．04％：作用时间48h，各浓度腺苷对S65细胞的抑制率分别为11．43％、23．10％、34．76％和46．67％；随着腺苷浓度的增高，S65细胞的增殖受到明显的抑制，呈现明显的浓度效应依赖关系。而腺苷对U95细胞的增殖无明显影响。结论腺苷能够明显抑制葡萄膜黑色素瘤细胞的增殖，但不影响正常葡萄膜黑色素细胞的生长。腺苷和／或腺苷受体激动剂也许可以作为一种新的药物用于葡萄膜黑色素瘤的治疗。

Effect of adenosine on cell proliferation in uveal melanocytes and uveal melanoma cells

Objective To investigate expression of adenosine receptors in uveal melanocytes and uveal melanoma cell, and the effect of adenosine on cell proliferation on these two types of cells. Methods Adenosine receptors (A1, A2A, A2B, A3) mRNA expression were detected in cultured uveal melanocytes cell U95 and uveal melanoma cell S65 by Real-time PCR. After U95 and S65 cells were treated with 0, 0.1, 1, 10 or 100 μLmol/L adenosine for 24 h or 48 h, cell viability of each group was measured by MTT. Results Four subtypes of adenosine receptors (A1, A2A, A2B and A3) mRNA were expressed in U95 and S65 cells. The expression of adenosine receptors A1 and A2B is similar to each other but higher than that of A2A and A3 in U95 cells (A1: 1.32±0.47; A2A: 0.28± 0.05; A2B: 1.03±0.10; A3: 0.20±0.07; F=15.90, P<0.01). The mRNA expression of adenosine receptor A2B was much higher than the three other subtypes of adenosine receptors in S65 cells (A1: 1.03±0.27; A2A: 2.23±0.35; A2B: 127.34±18.69; A3: 0.17±0.07; F:163.20, P<0.01). Adenosine receptor A2B and A2A mRNA expression was higher in S65 cells than in U95 cells (A2A: t=9.35, P< 0.01; A2B: t=11.43, P<0.01). After treated with 0, 0.1, 1, 10 and 100 μmol/L adenosine for 24 h and 48 h, S65 cells growth inhibition ratio were 3.47%, 14.93%, 19.79%, 26.04% at 24 h and 11.43%, 23.10%, 34.76%, 46.67% at 48 h. However different concentrations of adenosine had no effect on the proliferation of U95 cells. Conclusion The adenosine receptor subtypes express differently in uveal melanocytes and uveal melanoma cells. Adenosine is able to inhibit uveal melanoma cells proliferation, but has no effect on uveal melanocytes cells, which may be because of different adenosine receptor subtypes expression in these two types of cells.