低氘水对肺癌细胞增殖的抑制作用及其可能的机制

Inhibitory effect of deuterium-depleted water on proliferation of lung carcinoma cells and the possible mechanism

目的:初步探讨低氘水(deuterium-depleted water,DDW)在体内外对人肺癌细胞增殖的抑制作用及其可能机制。方法:MTT法检测DDW对肺癌A549细胞和正常人胚肺成纤维细胞HLF-1增殖的抑制作用,TUNEL法检测A549细胞凋亡情况,流式细胞仪检测细胞周期的变化。建立BALB/c裸鼠人肺癌H460细胞移植瘤模型,低氘水饮用60d后观察裸鼠移植瘤的生长情况。结果:与对照组比较,培养10h时体积分数为0.0025%、0.0050%和0.0105%的DDW对A549细胞增殖具有显著抑制作用(P<0.05),随后抑制作用消失,48h后又逐渐出现抑制现象,72h抑制作用显著(P<0.05);相同条件下不同体积分数的DDW对正常人胚肺成纤维细胞HLF-1无显著抑制作用。TUNEL染色显示,0.005%DDW作用后A549细胞出现凋亡,凋亡率显著高于对照组细胞[(25.38±3.90)%vs(10.87±1.11)%),P<0.05]。流式细胞仪检测显示,DDW作用后A549细胞S期细胞显著增加(P<0.05)。人肺癌细胞H460移植瘤裸鼠饮用DDW后,能够明显提高裸鼠的生活质量,抑瘤率达30.08%。结论:DDW对肺癌细胞增殖的抑制作用仅限于一定的剂量范围,且具有波动性和时段性的特点;其机制可能与诱导肺癌细胞S期阻滞和细胞凋亡有关。

Objective：To explore the in vitro and in vivo inhibitory effects of deuteriumdepleted water （DDW） on the proliferation of human lung carcinoma cells, and to explore the possible mechanism. Methods： The inhibitory effect of DDW on the proliferation of human lung carcinoma A549 cells and human embryonic lung fibroblast HLF1 cells was examined by MTT assay; apoptosis of A549 cells was examined by TUNEL; and cell cycle was analyzed by flow cytometry. Mouse model of lung carcinoma was established by inoculating human lung carcinoma H460 cells into BALB/c nude mice, and the growth of implanted tumors was observed after DDW treatment for 60 d. Results： Compared with control group, A549 cells treated with 0.0025%, 0.0050% or 0.0105% DDW showed significantly decreased proliferation 10 h after treatment （P〈0.01）. Then the inhibitory effects of DDW gradually disappeared, but appeared 48 h later again, with the inhibitory effects at 72 h being significant （P〈0.05）. DDW at the same dosages showed no inhibition on the proliferation of HLF1 cells （P〉0.05）. TUNEL assay verified the apoptosis of DDWtreated A549 cells, and the apoptosis rate of DDWtreated A549 cells was significantly higher than that of control group（[45.30±4.21]% vs [22.25±0.30]%, P〈0.01）. Cells in S phase were significantly increased in DDWtreated A549 cells compared with those in the control group （P〈0.05）. The life quality of H460 cellinoculated nude mice treated with DDW was greatly improved, with the tumor inhibition rate being 30.08%. Conclusion： DDW can inhibit the proliferation of lung cancer cells within a certain range of dosage and in a fluctuating pattern; its mechanism might be associated with induction of apoptosis and cell cycle arrest in S phase of tumor cells.