As_2O_3对体外肠癌细胞HCT116生长和增殖周期的影响

Effect of arsenic trioxide on growth and cell cycle progression in colon cancer cell line HCT116

目的:研究三氧化二砷(As2O3)对人体外肠癌细胞HCT116的生长抑制作用和增殖周期的影响.方法:肠癌细胞HCT116进行体外培养后,分成对照组,0.5μmol/L As2O3组、1.5μmol/L As2O3组、2.5μmol/L As2O3组,在不同时间段内,MTT法观察不同浓度As2O3对肠癌细胞的生长抑制情况,绘制细胞生长曲线;流式细胞技术分析细胞增殖的周期变化.结果:MTT结果显示0.5μmol/L的低剂量A s2O3抑制率与对照组无明显差别(P>0.05),1.5μmol/L As2O3组抑制率和2.5μmol/L As2O3组抑制率与对照组相比较,组间均有差异(P<0.05),且As2O3对肿瘤细胞的抑制作用均随时间延长而增强,并在第3天时抑制作用最明显(1.5μmol/L As2O3组抑制率为38.64%±0.16%,2.5μmol/L As2O3组抑制率为51.42%±0.53%,对照组是8.35%±0.76%),然后第4天有下降的趋势,但2.5μmol/L As2O3的抑制作用第4天未见明显下降趋势(抑制率为52.93%±1.53%);流式细胞术分析表明,As2O3为0.5μmol/L时,S期细胞分布35.58%±0.63%(对照组25.69%±1.46%),G2/M期细胞分布33.41%±0.73%(对照组30.44%±1.51%)两者比较没有统计学差异(P>0.05),当As2O3浓度为1.5μmol/L时,S期细胞占42.69%±2.64%,G2/M期细胞占22.46%±0.59%,与对照组比较,差异有统计学意义(P<0.05);药物提高到2.5μmol/L时,S期细胞占45.71%±1.53%,G2/M期细胞占14.66%±0.92%,与对照组比较差异有统计学意义(P<0.05).结论:As2O3对人肠癌细胞HCT116具有明显的抑制作用,主要抑制肿瘤细胞DNA合成的增殖期,且至少1.5μmol/L以上的As2O3浓度才能达到抑制肿瘤增长繁殖的效果,所以临床应用As2O3进行肠癌化疗时应掌握好有效的剂量浓度和化疗时间窗,从而提高化疗效果的同时最大减轻不良反应,延缓耐药性的发生.

AIM： To study the effect of arsenic trioxide （As203） on cell growth and cell cycle progression in human colon carcinoma cell line HCT116.METHODS： HCT116 cells cultured in vitro were divided into a control group, a 0.5 μmol/L AS203 group, a 1.5 μmol/L AS203 group, and a 2.5 pmol/L As203 group. After treatment with As203 for different durations, the effect of dif- ferent concentrations of arsenic trioxide on the growth of colon cancer cells was detected by MTT assay. Cell growth curve was plotted to observe the change in cell proliferation. Flow cy- tometry （FCM） was used to determine cell cycle progression.RESULTS： Low dose of As203 （0.5 ~mol/L） had no significant inhibitory effect on HCT116 cells compared with the control group （P 〉 0.05）, but 1.5 and 2.5 ~tmol/L of As203 showed a signifi- cant inhibitory effect on cell growth in a time- dependent manner （38.64% ± 0.16%, 51.42% ± 0.53% vs 8.35% ± 0.76%, P 〈 0.05 for both）. In the 1.5 ~tmol/L As203 group, the inhibitory ef- fect was most obvious on day 3, then gradually declined; however, such a downward trend was not observed in the 2.5 ~tmol/L As203 group （52.93% ± 1.53%）. FCM analysis showed that AS203 at a concentration of 0.5 μmol/L had no significant effect on the percentages of cells in S phase and G2/M phase （35.58% ± 0.63% vs 25.69% ± 1.46%; 33.41% ± 0.73% vs 30.44% ± 1.51%, P 〉 0.05 for both）. However, AS203 at concentrations of 1.5 ~mol/L and 2.5 i^mol/L significantly increased the percentages of cells in S phase but decreased the percentages of cells in G2/M phase （1.5 ~mol/L： 42.69% ± 2.64% and 22.46% ± 0.59%; 2.5 t^mol/L： 45.71% ± 1.53% and 14.66% ± 0.92%; P 〈 0.05 for all）. CONCLUSION： Arsenic trioxide has an obvious inhibitory effect on the proliferation of HCT116 cells, mainly by inhibiting the synthesis of DNA in the proliferation stage. The effective dose and treatment time are important in clinical applica- tion of As203 for cancer chemotherapy in order to improve the effect of chemotherapy, reduce toxicity reactions and delay the occurrence of drug resistance.