复方二氯乙酸二异丙胺对食管癌的抗肿瘤活性及药理学机制研究

Antitumor Activity and Pharmacological Mechanism of Compound Dichloroacetic Acid Diisopropylamine on Esophageal Cancer

目的探讨复方二氯乙酸二异丙胺(DADA)对食管癌的抗肿瘤活性及药理学机制。方法购买食管鳞癌细胞株Eca-109和TE-13,制作大鼠皮下荷瘤模型,使用染色流式细胞技术及Western blot检测方法、活性氧(ROS)试剂检测盒检测食管癌的细胞株内γ-H2AX的表达、细胞周期、细胞凋亡及ROS水平。结果 DADA对Eca-109和TE-13细胞株的抗肿瘤活性高于二氯乙酸(DCA);经DADA处理后的Eca-109和TE-13细胞株通过小室膜的细胞数目大量减少,细胞内的Cleaved Caspase-3蛋白含量显著增加(P<0.05);DADA联合放射治疗(简称放疗)较单纯放疗食管鳞癌Eca-109和TE-13细胞株敏感度分别增加1.38倍和1.05倍;DADA联合放疗与单用DADA、单用放疗相比,Eca-109和TE-13细胞株中γ-H2AX蛋白水平显著升高,G2/M周期的细胞数量显著升高,ROS水平显著增加(P<0.05);大鼠体内验证结果显示,DADA联合放疗治疗大鼠肿瘤体积及质量均显著低于单用DADA及单用放疗大鼠(P<0.05)。结论 DADA可抑制食管癌肿瘤细胞生长,促进凋亡并抑制其迁移,与放疗联用可通过增加癌细胞中ROS水平而增加食管癌放疗的敏感性,从而更好地发挥其抗肿瘤活性。

Objective To investigate the antitumor activity and pharmacological mechanism of compound dichloroacetic acid diisopropylamine（DADA） on esophageal cancer. Methods The esophageal squamous carcinoma cell lines Eca- 109 and TE- 13 were purchased for the subcutaneous tumor- bearing models of rats. The expression of γ-H2AX,cell cycle,apoptosis and ROS levels in cell lines of esophageal cancer were detected by staining and flow cytometry, Western blot detection method, and reactive oxygen species（ROS） detection kit. Results The antitumor activity of DADA on Eca- 109 and TE- 13 cell lines was higher than that of DCA. The number of cells passing through the ventricular membranes of DADA-treated Eca- 109 and TE- 13 cell lines were significantly reduced,the content of Cleaved Caspase-3 protein in cells were increased significantly（P 〈 0.05）. Compared with radiotherapy alone,the sensitivity of esophageal squamous cell carcinoma Eca- 109 and TE- 13 cell lines increased by 1.38 times and 1.05 times respectively afterthat the levels of γ- H2AX protein in the Eca DADA group and radiotherapy group,DADA combined with radiotherapy group showed 109 and TE- 13 cell lines were significantly increased,the number of cells in G2/M cycle was significantly increased and the level of ROS was significantly increased（P 〈 0. 05）. The results of in vivo validation showed that the tumor volume and weight of DADA combined with radiotherapy were significantly lower than those of DADA or radiotheraW alone（P 〈 0.05）. Conclusion DADA can inhibit the growth of esophageal cancer cell, promote apoptosis and inhibit its migration, combined with radiotherapy can increase the ROS level in cancer cells,and increase the sensitivity of esophageal cancer to radiotherapy, so as to better exert its antitumor activity.