达肝素钠抑制人肺腺癌细胞增殖作用的实验观察
Experimental study of the role of dalteparin sodium in inhibiting the proliferation of human lung adenocarcinoma cell
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摘要: 目的:探讨达肝素钠抑制人肺腺癌细胞增殖的作用机制。方法:采用流式细胞术检测达肝素钠处理后A549细胞的细胞周期变化。建立肺腺癌鼠模型后,将12只荷瘤鼠随机分为达肝素钠组和对照组,分别给予达肝素钠1 500 IU/kg(0.2 ml)皮下注射和0.9%氯化钠注射液0.2 ml腹腔注射,1天1次,连续用药35 d。定期测量瘤体最大及最小直径,计算瘤体积、绘制肿瘤生长曲线。第35天用戊苯巴比妥钠全身麻醉处死裸鼠,剥取瘤体称重、计算抑瘤率,并用免疫组织化学方法测瘤组织中细胞周期蛋白E(Cyclin E)的表达。结果:肺腺癌细胞株A549处理48 h后,与对照组比较,达肝素钠组G0/G1期细胞比例升高,S期细胞比例降低(P0.01);与对照组比较,达肝素钠组瘤重明显减小(P0.05);达肝素钠组抑瘤率为0.000%和64.081%;对照组及达肝素钠组瘤组织中Cyclin E的表达阳性率为6/6、3/6,差异无统计学意义(P=0.182);达肝素钠组主要表现为低表达(5/6),对照组为高表达(5/6),差异无统计学意义(P=0.081)。结论:达肝素钠可阻滞人肺腺癌A549细胞于G0/G1期,使进入S期和M期的细胞减少,减少DNA的合成,抑制细胞增殖,此作用可能与其抑制细胞周期调控因子Cyclin E的表达有关。Abstract: Objective:To investigate the mechanism of dalteparin in inhibiting the human lung adenocarcinoma cell proliferation． Methods:The cycle changes of A549 cells after dalteparin treatment were detected by flow cytometry． After the mouse models of lung cancer were established，the 12 tumor-bearing mice were randomly divided into dalteparin group and control group;they were given 1 500 IU/kg(0． 2 ml) subcutaneously or 0． 9% chloride sodium intraperitoneally(0． 2 ml) once a day for 35 days． The maximum diameter and the minimum diameter of the tumor were measured regularly;the tumor volume was calculated and the tumor growth curve was made． Thirty-five days later，the mice were executed after anesthetized with amyl phenobarbital sodium． The tumor was stripped and weighed，the inhibitory rate was calculated and the expression of Cyclin E in tumor tissue was measured using immunohistochemical method． Results:Forty-eight hours after the lung adenocarcinoma cell line A549 was treated，the proportion of cells in G0/G1phase were increased and S phase were decreased in the dalteparin group compared with that of the control group(P 0． 01);the tumor weight in the dalteparin group was significantly reduced compared with that in the control group(P 0． 05);the tumor inhibition rate in the dalteparin group was 64． 081%． The positive expression of Cyclin E was 6/6 and 3/6 respectively in the dalteparin group and the control group;the difference was not statistically significant(P = 0． 182)． The dalteparin group mainly showed low expression(5/6)， while the control group showed high expression(5/6);the difference was not statistically significant(P = 0． 081)． Conclusions: Dalteparin can block human lung adenocarcinoma A549 cells in G0/G1phase，the cell entry into S phase and M phase is reduced，the DNA synthesis is decreased and the cell proliferation is inhibited，which might be related to the expression of inhibiting cell cycle regulator Cyclin E．
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Key words:
- lung neoplasms /
- adenocarcinoma /
- daltepain /
- cell proliferation /
- Cyclin E /
- nude mice
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