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目的:研究全反式维甲酸(ATRA)联合替莫唑胺(TMZ)对胶质瘤细胞株U251增殖及凋亡的影响。方法:体外培养胶质瘤细胞株U251,分为ATRA组、TMZ组、ATRA+TMZ组和空白对照4组,应用MTT法测定U251细胞生长抑制率,流式细胞仪检测细胞周期分布和细胞凋亡率,western blot法检测细胞维甲酸受体β(RARβ)蛋白和凋亡相关蛋白Caspase-3蛋白的表达情况。结果:对照组、ATRA组、TMZ组及ATRA+TMZ组的细胞生长抑制率分别为(1.72±0.12)%、(9.87±0.87)%、(23.87±1.32)%及(35.74±1.44)%,ATRA+TMZ组的细胞生长抑制率显著高于单独应用TMZ(P<0.01)。对照组、ATRA组、TMZ组及ATRA+TMZ组的细胞凋亡率分别为(1.32±0.11)%、(4.16±0.35)%、(8.44±0.49)%、(15.27±1.03)%,ATRA+TMZ组的凋亡率显著高于单独应用TMZ有更高的凋亡率(P<0.01)。对照组、ATRA组、TMZ组及ATRA+TMZ组RARβ蛋白相对表达量分别0.452±0.054、0.837±0.068、0.195±0.021、0.376±0.039,ATRA+TMZ组RARβ蛋白相对表达量显著高于TMZ组(P<0.01)。ATRA+TMZ组Caspase-的3蛋白表达相对水平为(0.832±0.059),明显高于ATRA组(0.334±0.041)及TMZ组(0.521±0.032),差异具有统计学意义(P<0.01)。结论:全反式维甲酸联合替莫唑胺能更有效抑制U251细胞的增殖,增加其凋亡率,这可能与其增加RARβ和Caspase-3蛋白的表达抑制U251细胞增殖、诱导细胞凋亡有关。
Objective: To investigate the effects of all-trans retinoic acid (ATRA) combined with temozolomide (TMZ) on the proliferation and apoptosis of glioma cell line U251. Methods: Glioma cell line U251 was cultured in vitro. The cells were divided into four groups: ATRA group, TMZ group, ATRA + TMZ group and blank control group. The growth inhibition rate of U251 cells was determined by MTT assay. Cell cycle distribution and apoptosis were detected by flow cytometry The protein expression of RARβ and Caspase-3 were detected by Western blot. RESULTS: The cell growth inhibition rates in control, ATRA, TMZ and ATRA + TMZ groups were (1.72 ± 0.12)%, (9.87 ± 0.87)%, (23.87 ± 1.32)% and (35.74 ± 1.44)%, The cell growth inhibition rate of ATRA + TMZ group was significantly higher than TMZ alone (P <0.01). The apoptotic rates of ATRA group, ATRA group, ATRA + TMZ group were (1.32 ± 0.11)%, (4.16 ± 0.35)%, (8.44 ± 0.49)%, (15.27 ± 1.03)%, The apoptosis rate of TMZ group was significantly higher than that of TMZ alone (P <0.01). The relative expression of RARβ protein in ATRA group, TMZ group and ATRA + TMZ group were 0.452 ± 0.054,0.837 ± 0.068,0.195 ± 0.021,0.376 ± 0.039, respectively. The relative expression of RARβ protein in ATRA + TMZ group was significantly higher than that in TMZ group P <0.01). The relative expression of Caspase-3 in ATRA + TMZ group was (0.832 ± 0.059), which was significantly higher than that in ATRA group (0.334 ± 0.041) and TMZ group (0.521 ± 0.032) (P <0.01). CONCLUSION: All-trans retinoic acid combined with temozolomide can effectively inhibit the proliferation and increase the apoptosis rate of U251 cells, which may be related to the increase of the expression of RARβ and Caspase-3 protein, and the inhibition of U251 cell proliferation and apoptosis induction.