双硫仑对人脂肪肉瘤细胞系SW872细胞的抑制作用及其机制

Suppressive effect of disulfiram on growth of liposarcoma cell line SW872 and its mechanisms

目的观察双硫仑(DSF)对人脂肪肉瘤细胞SW872的抑制作用,并探讨其相关分子机制。方法体外培养SW872细胞,设立正常细胞对照组及DSF 1,2.5,5和10μmol·L-1组,采用CCK-8法检测药物处理24 h后对细胞增殖的抑制作用;光镜下观察DSF处理后SW872细胞形态变化;流式细胞术检测细胞凋亡;克隆形成实验检测DSF对SW872细胞克隆形成能力的影响;Western印迹法检测A20的表达;CCK-8法检测铁离子螯合剂Fer-1及炎症小体NLRP3抑制剂MCC950能否逆转DSF对SW872细胞的抑制作用;RT-PCR法检测DSF对SW872细胞中A20和醛脱氢酶(ALDH1)的mRNA水平。结果 DSF对SW872细胞增殖有显著抑制作用(P<0.01);DSF处理后使SW872细胞发生皱缩变圆且细胞间隙增大;DSF作用于SW872细胞24 h后,与正常细胞对照组相比,DSF 1μmol·L-1组细胞早期凋亡率明显增加〔(32.6±1.82)%vs(3.50±0.64)%,P<0.05〕;DSF 0.1μmol·L-1组明显抑制SW872细胞的克隆形成〔(16.7±7.02)%vs(129±5.29)%,P<0.05〕;DSF处理能够明显提高SW872细胞中炎症调节分子A20的表达;铁离子鳌合剂Fer-1不能逆转DSF导致的细胞生长抑制,而炎症小体抑制剂MCC950可以部分逆转DSF对SW872细胞增殖的抑制作用。结论 DSF能抑制SW872细胞增殖并诱导其凋亡,其机制可能通过细胞炎症小体介导,细胞炎症调节分子A20参与DSF对SW872细胞的抑制作用。

OBJECTIVE To observe the inhibitory effect of disulfiram(DSF)on proliferation of human liposarcoma cell line SW872 and to explore its related molecular mechanisms.METHODS SW872 cells were cultured in DSF at different concentrations.CCK-8 assay was used to detect the inhibitory effect of DSF on cell proliferation.Apoptosis was detected by flow cytometry.A20 was detected by RT-PCR and Western blotting.CCK-8 assay was used to find out whether Fer-1 or MCC950 could reverse the inhibitory effect of DSF on SW872 cell growth.ALDH1A1-3 mRNA of SW872 cells was detected by RT-PCR.RESULTS DSF significantly inhibited the proliferation of SW872 cells(P<0.01).DSF treatment caused SW872 cells to shrink and turn round while increasing the intercellular space.The apoptosis rate of SW872 cells treated by DSF was significantly increased compared with control group.DSF inhibited the colony formation ability of SW872 cells.Furthermore,A20 was significantly increased in SW872 cells treated with DSF.MCC950 could reverse the inhibitory effect of DSF on SW872 cell growth,but Fer-1 could not,suggesting that NLRP3 inflammasome be involved in DSF induced growth inhibition.DSF treatment induced the ALDH1A1-3 expression which reached the highest level at the concentration of 5μmol·L-1.CONCLUSION DSF can inhibit the proliferation and induce apoptosis of human liposarcoma SW872 cells.The mechanism may be revealed that A20 is involved in DSFinduced growth inhibition which is not through ferroptosis.