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目的:研究松油烯-4-醇(T4O)对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T4O体外作用WM35细胞。CCK-8测抑制率;AO/EB染色后倒置显微镜观察细胞形态;比色法测Caspase-3酶活性;流式细胞术测凋亡及周期;JC-1测线粒体膜电位(MMP);蛋白印迹测Bax及Bcl-2表达。结果:T4O以时间-剂量依赖性方式抑制WM35细胞增殖。T4O诱导WM35细胞凋亡,呈剂量依赖性,随药物浓度增加,Caspase-3酶活性逐渐增强,MMP逐渐降低,Bax/Bcl-2比值升高,周期阻滞在G2/M期。结论:T4O对WM35细胞有抑制增殖及促凋亡作用,其机制是通过线粒体途径使凋亡相关蛋白Bax表达上调、Bcl-2下调,激活凋亡途径关键酶Caspase-3,使细胞周期阻滞在G2/M期。
Objective: To study the effect of terpinen-4-ol (T4O) on the proliferation and apoptosis of WM35 cells and to explore its mechanism. Methods: Different concentrations of T4O in vitro effects of WM35 cells. CCK-8 measured inhibition rate; AO / EB staining inverted microscope to observe the cell morphology; Caspase-3 activity by colorimetric assay; Flowcytometry apoptosis and cycle; JC-1 measured mitochondrial membrane potential (MMP) The expression of Bax and Bcl-2 were detected by blotting. Results: T4O inhibited WM35 cell proliferation in a time-and dose-dependent manner. T4O induced WM35 cell apoptosis in a dose-dependent manner. With the increase of drug concentration, the activity of Caspase-3 gradually increased, the MMP decreased, the ratio of Bax / Bcl-2 increased, and the cycle arrest was at G2 / M phase. Conclusion: T4O can inhibit the proliferation and induce the apoptosis of WM35 cells through up-regulating the expression of Bax, down-regulating Bcl-2 and activating the key enzyme Caspase-3 in the apoptosis pathway through mitochondrial pathway, In G2 / M period.