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[目的]研究赖氨匹林(aspisol)对人恶性黑色素瘤A375细胞凋亡的诱导作用,并初步探讨其诱导A375细胞凋亡的可能机制。[方法]采用四甲基偶氮唑蓝(MTT)比色法检测赖氨匹林对A375细胞增殖的抑制作用;采用吖啶橙(AO)/溴化乙啶(EB)双染法进行A375细胞形态学观察;流式细胞仪(FCM)检测不同浓度的赖氨匹林诱导A375细胞的早期凋亡率;应用Westernblot法检测凋亡相关蛋白Bcl-2、Bax的表达。[结果]⑴不同浓度的赖氨匹林对A375细胞均有明显的生长抑制作用,且其作用具有时间和浓度相关性(P<0.01)。⑵AO/EB双染后可见:给药组早期凋亡细胞多于对照组,并且随着赖氨匹林浓度的增加,早期凋亡细胞和晚期凋亡细胞均增多。⑶不同浓度的赖氨匹林对A375细胞作用24h后,均可诱导其发生早期凋亡,并且呈浓度相关性。⑷赖氨匹林可上调Bax蛋白的表达,下调Bcl-2蛋白的表达。[结论]赖氨匹林可以抑制A375细胞的增殖,诱导其凋亡,此作用可能与调节Bax/Bcl-2蛋白的表达水平有关。
[Objective] To investigate the induction effect of aspisol on the apoptosis of human melanoma A375 cells and to explore its possible mechanism of inducing A375 cell apoptosis. [Method] MTT assay was used to determine the inhibitory effect of lysipirin on the proliferation of A375 cells. A375 was stained with acridine orange (AO) / ethidium bromide (EB) Cell morphology was observed. Flow cytometry (FCM) was used to detect the early apoptotic rates of A375 cells treated with different concentrations of 17, and Western blot was used to detect the expression of Bcl-2 and Bax. [Results] (1) Different concentration of meticlopram significantly inhibited the growth of A375 cells, and the effect was correlated with time and concentration (P <0.01). ⑵ AO / EB double staining shows: the early apoptotic cells in the treatment group more than the control group, and with the increase of the concentration of lysiplatin, early apoptotic cells and late apoptotic cells were increased. (3) Different concentration of methamidophos on A375 cells after 24h, can induce its early apoptosis, and showed a concentration-related. Â ’£ lysicillin can up-regulate the expression of Bax protein, down-regulate Bcl-2 protein expression. [Conclusion] L-arginine can inhibit the proliferation and induce the apoptosis of A375 cells, which may be related to the regulation of Bax / Bcl-2 protein expression.