来氟米特对前列腺癌PC3细胞增殖与凋亡的影响

The effects of Leflunomide on cellular proliferation and apoptosis in PC3 cells

为了研究来氟米特(LEF)对前列腺癌细胞增殖和凋亡的影响,采用前列腺癌细胞株PC3与不同浓度的来氟米特(0,50,100,150μmol·L^(-1))共培养(24,48,72 h),通过CCK-8细胞活力检测实验和克隆形成实验,发现来氟米特抑制PC3细胞增殖,且存在明显的浓度时间依赖性;流式细胞检测表明,来氟米特使PC3细胞发生S期阻滞,并且促进其凋亡;当来氟米特浓度≥150μmol·L^(-1)时,Western Blot实验检测到Caspase-3蛋白发生剪切活化.来氟米特处理可诱导PC3细胞自噬,采用羟氯喹抑制自噬可促进细胞凋亡的发生,提示自噬增强了细胞抗凋亡能力.研究表明,来氟米特可抑制PC3细胞的增殖,诱导其凋亡,可能对前列腺癌有潜在的治疗作用.

Leflunomide （LEF）, an immunosuppressive drug, was generally used in the treatment of rheumatoid arthritis. Recently, increasing evidences indicated that LEF had an effective anti-proliferative effect on various tumors. To investigate the effect of LEF in prostatic cancer cells, prostatic cancer cell line PC3 was trea-ted with solutions of LEF （0, 50, 100 and 150 μmol·L^-1） and cultured in varied duration of time （24, 48 and 72 h）. CCK-8 and cell clone forming experiments showed that when the concentration was ≥ 150 μmol·L^-1 LEF significantly inhibited cell growth in a time- and dose-dependent manner. Annexin V-FITC/PI staining assay revealed that LEF induced S-phase cell cycle arrest, and enhanced cell apoptosis via modulation of Caspase-3 expression as shown by Western blotting experiments. Moreover, LEF could induce autophagy of PC3 cells, while hydroxychloroquine （HCQ） could enhance LEF-induced apoptosis by inhibiting autophagy. Taken together, LEF inhibited proliferation and promoted apoptosis of the PC3 cells.