α-硫辛酸增强吡咯烷二硫代氨基甲酸对 Hep-2细胞增殖的抑制作用

摘要

目的：探讨α-硫辛酸（α-LA）对吡咯烷二硫代氨基甲酸（PDTC）抑制 Hep-2细胞增殖的影响。方法体外培养人喉癌细胞系 Hep-2细胞，加入α-LA、PDTC 或α-LA 联合 PDTC 作用12，24和48 h 后，用 WST-1法和软琼脂集落形成法检测细胞增殖，Hoechst33258染色法和流式细胞仪检测细胞凋亡和细胞周期。结果α-LA 5~500μmol·L-1对 Hep-2细胞增殖无抑制作用，PDTC 5~50μmol·L-1能明显抑制Hep-2细胞增殖（P﹤0.05，P﹤0.01），PDTC 5μmol·L-1联合α-LA 5，10和20μmol·L-1对 Hep-2细胞增殖的抑制率明显高于对照组（P﹤0.01）和 PDTC 单独处理组（P﹤0.05）。PDTC 5μmol·L-1与α-LA 5μmol·L-1联合用药，在12~48 h 内对 Hep-2细胞增殖的抑制率呈时间依赖性（r ＝0.987，P﹤0.05），24 h 后软琼脂细胞集落形成数与 PDTC 单独处理（P﹤0.05）和α-LA 单独处理组相比明显减少（P﹤0.01）。Hoechst33258染色和流式细胞仪检测结果表明，与 PDTC 和α-LA 单独处理组比较，PDTC 5μmol·L-1与α-LA 5μmol·L-1联合用药24 h 凋亡细胞增加，G2/ M 期细胞百分率增加（P﹤0.05）。结论α-LA 可增强 PDTC 对 Hep-2细胞增殖的抑制作用并诱导 Hep-2细胞凋亡。%OBJECTIVE To investigate whether α-lipoic acid(α-LA)can promote the anti-prolifera-tion effect of pyrrolidine dithiocarbamic acid(PDTC)in Hep-2 cells. METHODS The laryngeal carcino-ma Hep-2 cells were cultured with α-LA,PDTC or α-LA+PDTC respectively for 12,24 and 48 h. The proliferation of Hep-2 cells was detected by WST-1 assay and soft agar colony formation while apoptosis and cell cycle were analyzed by Hoechst33258 staining and flow cytometry. RESULTS α-LA 5 -500 μmol·L-1 could not inhibit Hep-2 cell proliferation,but PDTC 5 -50 μmol·L-1 could( P ﹤0.05,P ﹤0.01). The cell proliferation inhibitory rate of PDTC 5 μmol·L-1 combined with α-LA 5,10 and 20 μmol·L-1 groups was much higher than in control group(P﹤0.01)and PDTC alone group(P﹤0.05). When α-LA 5 μmol·L-1 was used in combination with PDTC 5 μmol·L-1 for 12-48 h,the cell proliferation was inhibited in a time-dependent manner(r=0.987,P﹤0.05). When the cells were treated for 24 h,the number of soft agar colony formations in combined group was significantly smaller than that of both α-LA alone group(P﹤0.01)and PDTC alone group(P﹤0.05). The result of Hoechst33258 staining and flow cytometry indicated that after combined treatment with PDTC 5 μmol·L-1 and α-LA 5 μmol·L-1 for 24 h,the level of apoptosis and the percentage of cells in G2 / M stage were significantly increased compared with PDTC alone or α-LA alone treatment. CONCLUSION α-LA can enhance the anti-proliferation and pro-apoptosis effect of PDTC in Hep-2 cells.