阿达帕林对皮肤T细胞淋巴瘤细胞增殖、凋亡及周期的影响

Effects of adapalene on the proliferation,apoptosis and cell cycle distribution of cutaneous T-cell lymphoma cells

目的探讨阿达帕林对人皮肤T细胞淋巴瘤(CTCL)Hut78细胞增殖、凋亡及周期的影响。方法采用CCK-8法检测不同浓度(0、0.01、0.1、1、2.5、5、10、25、50和100μmol/L)阿达帕林对人CTCL Hut78、Myla和HH细胞的增殖抑制作用,并分别计算IC_(50)。采用膜联蛋白(Annexin)Ⅴ/碘化丙啶(PI)双染法和PI单染法分别检测阿达帕林0、2、4和6μmol/L对Hut78细胞凋亡和周期分布的影响,Western blot法检测阿达帕林对Hut78细胞DNA损伤标志蛋白γ-H2AX表达的影响。结果阿达帕林对Hut78、Myla和HH细胞增殖的抑制作用具有浓度依赖性,Hut78细胞的IC_(50)为(1.45±0.47)μmol/L,HH细胞的IC_(50)为(2.89±0.22)μmol/L,Myla细胞的IC_(50)为(30.51±2.12)μmol/L。与阿达帕林0μmol/L组相比,阿达帕林2、4和6μmol/L组Hut78细胞的凋亡率升高[(10.82±0.46)%、(55.51±2.88)%、(73.26±2.21)%vs.(5.79±0.90)%](P<0.01),S期细胞比例升高[(72.51±1.89)%、(69.84±1.89)%、(91.68±10.84)%vs.(45.99±0.48)%](P<0.01),阿达帕林4和6μmol/L组细胞中γ-H2AX表达增加(P<0.05)。结论阿达帕林可选择性抑制CTCL中Hut78细胞的增殖,并诱导Hut78细胞凋亡和S期阻滞,其机制可能与阿达帕林诱导的DNA损伤有关。

Objective To investigate the effects of adapalene on the proliferation,apoptosis and cell cycle distribution of human cutaneous T-cell lymphoma Hut78 cells.Methods After cutaneous T cell lymphoma cells(Hut78,Myla and HH cells)were treated with adapalene in different concentrations of 0,0.01,0.1,1,2.5,5,10,25,50 and 100μmol/L for 72 hours,CCK-8 assay was performed to detect the inhibitory effects of adapalene on the cell proliferation and the IC50 was calculated.Subsequently,AnnexinⅤ/PI double staining and PI single staining were used to detect the effects of adapalene 0,2,4,6μmol/L(groups of A,B,C,D)on the apoptosis induction and cell cycle distribution in Hut78 cells,respectively.The expression of DNA damage biomarkerγ-H2 AX influenced by adapalene was evaluated by Western blot analysis.Results Adapalene inhibited the proliferation of human cutaneous T-cell lymphoma cells in concentration-dependent manner,and the IC50 of adapalene in Hut78,HH and Myla cells was(1.45±0.47)μmol/L,(2.89±0.22)μmol/L and(30.51±2.12)μmol/L,respectively.Compared with group A,the apoptosis rate of Hut78 cells and proportion of S-phase cells in groups of B,C and D were higher[(10.82±0.46)%,(55.51±2.88)%,(73.26±2.21)%vs.(5.79±0.90)%and(72.51±1.89)%,(69.84±1.89)%,(91.68±10.84)%vs.(45.99±0.48)%](P<0.01),and the expression ofγ-H2 AX in groups of C and D was increased(P<0.05).Conclusion Adapalene selectively inhibits the proliferation of Hut78 cells and triggers cell apoptosis and S phase arrest,which may be related to the DNA damage triggered by adapalene treatment.