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目的神经母细胞瘤(neuroblastoma,NB)是儿童常见的实体瘤,三氧化二砷(Arsenic troxide,As_2O_3)现已广泛应用于治疗实体肿瘤,为了探讨As_2O_3治疗NB的可行性,本研究检测As_2O_3对人SK-N-SH细胞周期及Trk基因家族表达的影响。方法通过流式细胞术检测4μmol/L浓度的As_2O_3作用48h后SK-N-SH细胞细胞周期的变化;qRT-PCR法检测As_2O_3在1、2和4μmol/L浓度下对SK-N-SH细胞分别作用24和48h后,对SK-N-SH细胞内TrkA、TrkB和TrkC基因表达的影响。结果用4μmol/L As_2O_3作用48h后,SK-N-SH细胞的G2/M期细胞比例为(30.93±0.34)%,而对照组G_2/M期细胞比例为(5.29±0.23)%,两组间差异有统计学意义,P<0.01。1μmol/L As_2O_3作用SK-N-SH细胞24h的TrkA比值为1.43±0.17,48h为3.88±0.26,两者比较差异有明显统计学意义,P<0.01,但随着As_2O_3浓度继续增大至4μmol/L TrkA反而下调。SK-N-SH细胞的TrkB及TrkC基因表达均随着As_2O_3浓度增加而上调,4μmol/L浓度下与对照组比较差异有统计学意义(P<0.05),48h比24h上调更明显。结论初步研究结果表明,As_2O_3不仅对NB的SK-N-SH细胞有明显杀伤作用,使其阻滞在G_2/M期,还能使与NB预后密切相关的Trk基因的表达发生变化,As_2O_3有用于NB治疗的潜在临床价值。
ObjectiveNoblastoma (NB) is a common solid tumor in children. Arsenic trioxide (As 2 O 3) has been widely used in the treatment of solid tumors. To investigate the feasibility of As 2 O 3 in the treatment of NB, this study examined the effect of As 2 O 3 on human SK- N-SH cell cycle and Trk gene family expression. Methods The cell cycle of SK-N-SH cells was detected by flow cytometry after treated with 4μmol / L As_2O_3 for 48h. The effect of As_2O_3 on the proliferation of SK-N-SH cells was detected by qRT-PCR at 1, 2 and 4μmol / The effects of TrkA, TrkB and TrkC on the expression of SK-N-SH cells were observed after 24 and 48 hours respectively. Results The percentage of cells in G2 / M phase of SK-N-SH cells was (30.93 ± 0.34)% after treated with 4μmol / L As 2 O 3 for 48h, while the proportion of cells in control group was (5.29 ± 0.23) There was significant difference between the two groups (P <0.01.1μmol / L As_2O_3), the TrkA ratio of SK-N-SH cells was 1.43 ± 0.17 at 24h and 3.88 ± 0.26 at 48h, the difference was statistically significant, P <0.01 , But down-regulated with increasing As 2 O 3 concentration to 4 μmol / L TrkA. The expression of TrkB and TrkC in SK-N-SH cells were up-regulated with the increase of As 2 O 3 concentration, and the difference was significant at 4 μmol / L compared with the control group (P <0.05). Conclusion Preliminary results show that As_2O_3 not only has a significant cytotoxic effect on NB-SK-N-SH cells, but also blocks the G_2 / M phase. It also changes the expression of Trk gene, which is closely related to the prognosis of NB, The potential clinical value of NB treatment.