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目的研究芳姜黄酮衍生物(ATD)对人皮肤黑色素瘤WM35细胞增殖及凋亡的影响,并探讨其作用机制。方法不同浓度(5~80μmol/L)ATD体外作用WM35细胞。CCK-8法检测增殖抑制率;AO/EB染色、倒置显微镜观察细胞凋亡形态;DNA片段化检测细胞凋亡;比色法检测Caspase-3酶活性;流式细胞术检测细胞凋亡;Western blotting检测Bax及Bcl-2蛋白表达。结果 ATD对WM35细胞有增殖抑制作用,呈时间-剂量依赖性(P<0.05)。ATD诱导WM35细胞凋亡,呈剂量依赖性(P<0.05),Caspase-3酶活性随药物浓度增加而增强(P<0.05)。随药物浓度增加,Bax/Bcl-2比值逐渐升高。结论 ATD对WM35细胞有抑制增殖及促凋亡作用,其机制是使凋亡相关蛋白Bax表达上调、Bcl-2表达下调,激活细胞凋亡途径关键酶Caspase-3,进而抑制肿瘤细胞分化与增殖。
Objective To study the effect of arginin derivative (ATD) on the proliferation and apoptosis of human cutaneous melanoma WM35 cells and to explore its mechanism. Methods WM35 cells were treated with ATD at different concentrations (5 ~ 80μmol / L) in vitro. The proliferation inhibition rate was detected by CCK-8 method. The apoptosis morphology was observed by AO / EB staining and inverted microscope. Apoptosis was detected by DNA fragmentation assay. Caspase-3 activity was detected by colorimetric assay. Apoptosis was detected by flow cytometry. blotting detection of Bax and Bcl-2 protein expression. Results ATD inhibited the proliferation of WM35 cells in a time-and dose-dependent manner (P <0.05). ATD induced WM35 cell apoptosis in a dose-dependent manner (P <0.05). The activity of Caspase-3 increased with the increase of drug concentration (P <0.05). With the increase of drug concentration, Bax / Bcl-2 ratio gradually increased. Conclusion ATD can inhibit the proliferation and induce the apoptosis of WM35 cells by down-regulating the expression of Bax, down-regulating the expression of Bcl-2 and activating the key enzyme Caspase-3 in the apoptosis pathway, thereby inhibiting the differentiation and proliferation of the tumor cells .