褪黑素促进Burkitt淋巴瘤细胞对阿霉素的化疗敏感性

Melatonin promotes chemotherapy sensitivity of Burkitt lymphoma cells to doxorubicin

目的探讨褪黑素(Melatonin,MEL)增强阿霉素(Adriamycin,ADM)抑制Burkitt淋巴瘤(Burkitt lymphoma,BL)细胞CA46的作用机制。方法使用不同浓度MEL(分为不加药组,0.5 mM组,1 mM组,2 mM组,4 mM组)分别处理CA466 h、12 h、24 h,使用ADM(分为不加药组,0.1μg/L组,0.2μg/L组,0.4μg/L组,0.8μg/L组,1.6μg/L组)处理CA4624 h,采用CCK8法检测细胞存活率并计算半数抑制率(half inhibition concentration,IC50)。同时,单用0.1μg/L ADM(ADM组),联用1 mM MEL和0.1μg/L ADM(AAM组)处理CA4624 h后,检测IC50的变化。使用蛋白印迹(Western Blotting,WB)实验检测不同浓度的MEL(分别为不加药组,0.5 mM组,1 mM组,2 mM组)处理CA4624 h后对NF-κB蛋白表达量的影响。WB法检测1 mM MEL处理CA4624 h(MEL组),ADM组,AAM组以及不加药组(Control组),WB法检测Nrf2,LC3B和p62蛋白的表达水平。结果MEL对CA46的抑制作用存在时间依赖性和浓度依赖性(P<0.05),同时降低了NF-κB蛋白的表达(P<0.05);同样ADM对CA46的抑制作用存在浓度依赖性(P<0.05);1 mM MEL处理CA466 h、12 h、24 h的IC50分别为(6.170±0.648)mM、(2.205±0.093)mM、(1.297±0.197)mM。ADM组和AAM组中ADM的IC50分别是(1.256±0.080)μg/L和(0.503±0.026)μg/L。AAM组相对于ADM组降低了Nrf2和p62的相对表达量(P<0.05),增强了LC3Ⅱ/LC3Ⅰ比值(P<0.05)。结论MEL增强ADM对CA46的抑制作用,其作用机制与抑制Nrf2/p62信号通路及增强自噬有关。

Objective To explore the mechanism by which melatonin(MEL)enhances the inhibition of CA46 in Burkitt lymphoma(BL)cells by adriamycin(ADM).Methods Different concentrations of MEL(no MEL treatment,0.5 mM MEL,1 mM MEL,2 mM MEL,4 mM MEL)were used to treat CA46 for 6 h,12 h,and 24 h,respectively.And ADM(no ADM treatment,0.1μg/L ADM,0.2μg/L ADM,0.4μg/L ADM,0.8μg/L ADM,1.6μg/L ADM)were treated with CA46 for 24 hours.The CCK8 method was used to detect the cell viability rate and calculate the half inhibition concentration(IC50).Meanwhile,CA46 was treated with 0.1μg/L ADM alone(ADM group)or 1 mM MEL combinded with 0.1μg/L ADM(AAM group)for 24 hours,and the IC50 was detected.Western Blotting(WB)analysis was used to detect the expression of NF-κB after different concentrations of MEL(no MEL treatment,0.5 mM MEL,1 mM MEL,2 mM MEL)treated on CA46 for 24 hours.WB was used to detect the expression levels of Nrf2,LC3 and p62 after 1 mM MEL treatment with CA46 for 24h(MEL group),ADM group,AAM group and Control group.Results The inhibitory effect of MEL on CA46 was time-dependent and concentration-dependent(P<0.05),and it also reduced the expression of NF-κB(P<0.05);similarly,the inhibitory effect of ADM on CA46 was concentration-dependent(P<0.05);the IC50 of CA46 treated with 1 mM MEL for 6 h,12 h and 24 h were(6.170±0.648)mM,(2.205±0.093)mM,and(1.297±0.197)mM,respectively.The IC50 of ADM in the ADM group and AAM group were(1.256±0.080)μg/L and(0.503±0.026)μg/L,respectively.Compared with the ADM group,the AAM group reduced the relative expression of Nrf2 and p62(P<0.05),and enhanced the LC3II/LC3I ratio(P<0.05).Conclusion MEL enhances the inhibitory effect of ADM on CA46,and its mechanism is related to the inhibiting of Nrf2/p62 signaling pathway and enhancing autophagy.