青蒿琥酯逆转裸鼠种植食管癌耐药作用研究

The resistance-reversal effect of artesunate on human esophageal cancer transplanted in nude mice

目的探讨三磷酸腺苷结合转运蛋白G2(ABCG2)与食管癌多药耐药的关系以及青蒿琥酯(Art)逆转食管癌多药耐药的作用机制。方法将食管癌多药耐药细胞Eca109/ABCG2接种于裸鼠皮下,建立荷瘤裸鼠动物模型。裸鼠皮下成瘤后用药,经腹腔给予Art和(或)阿霉素(ADM),分为Art高、低剂量组(50、25mg/kg Art),ADM组(1mg/kg ADM)和ADM+Art高、低剂量组(1mg/kg ADM+50、25mg/kg Art),对照组给予生理盐水。停药后第2天处死裸鼠,剥离瘤结节,网搓法制备单细胞悬液。采用流式细胞术检测皮下种植瘤细胞凋亡率,细胞ABCG2蛋白表达及细胞内ADM含量。结果成功建立Eca109/ABCG2种植瘤裸鼠模型,1周内成瘤率为100%。与对照组比较,ADM+Art高、低剂量组皮下种植瘤细胞凋亡率显著增加(P<0.05),细胞中ABCG2蛋白表达量显著降低(P<0.05),ADM含量显著增加(P<0.05)。结论 ABCG2参与了食管癌多药耐药的形成,Art可通过降低食管癌细胞中ABCG2的表达从而逆转食管癌耐药作用。

Objective To explore the relationship between ABCG2 and multidrug resistance of esophageal cancer and the mechanism of resistance-reversal effect by artesunate（Art）.Methods To establish the bearing cancer nude mice model by inoculating with Eca109/ABCG2 cells subcutaneously on the left subscapularis and study the resistance-reversal effect of artesunate on esophageal cancer using nude mice model.Injection drugs after subcutaneous tumor formation.Intraperitoneal injection with artesunate and adriamycin（ADM）.To set up Art high and low dose group（50,25mg/kg Art）,ADM group（1mg/kg ADM）,ADM＋ Art high and low dose group（1mg/kg ADM＋50,25mg/kg Art） and control group in the saline.The second day of the withdrawal of drugs,the nude mice were euthanized by breaking neck.The nodules of mice were stripped,then making into single cell suspension fluid with nets rub method.The apoptosis rate,ABCG2 protein expression level and ADM content in transplanted tumor cells were detected by flow cytometry.Results The model of esophageal cancer cell Eca109/ABCG2 xenograft in nude mice was successfully established,the tumorigenic rate was 100%.The growth inhibitory effect,ADM content in tumor cells and the apoptosis rate of esophageal cancer xenograft tumor cells treated with ADM in vivo were enhanced by combining artesunate without causing obvious side effects in treated mice.The ABCG2 protein expression level was significantly decreased in ADM＋ Art high and low dose group compared with ADM group（P0.05）.Conclusion ABCG2 participated in the development of esophageal cancer multidrug-resistance,artesunate could reverse the resistance of esophageal cancer by reducing the expression level of ABCG2.