AS_2O_3抑制人结肠癌生长的体内外实验研究

Study of growth inhibition by arsenic trioxide in human colon carcinoma cell

目的:通过研究三氧化二砷(AS2O3)对人结肠癌细胞株LS-174T在体内外实验中增殖情况的影响,探讨As2O3治疗结肠癌的机制。方法:1、试验分组:体外细胞培养及体内动物实验均分三组:对照组,As2O3低剂量组,As2O3高剂量组。2、MTT法测定细胞增殖数的情况。3、丫啶橙/溴乙啶(AO/EB)双荧光染色检测细胞凋亡率。4、免疫组织化学法测定结肠癌细胞和裸鼠实体肿瘤中HIF-1α和VEGF的表达。5、测量各组裸鼠实体肿瘤的重量及体积。结果:体外试验中,由MTT法可以看出,As2O3治疗组细胞生存率明显低于对照组(P<0.01),其中AS2O3高剂量组明显低于低剂量组。双荧光染色结果As2O3高剂量组细胞凋亡百分率明显高于低剂量组和对照组(P<0.01)。免疫组化结果显示结肠癌细胞HIF-1α和VEGF在对照组的阳性表达率明显高于As2O3治疗组(P<0.01),低剂量组高于高剂量组(P<0.05)。体内实验中,治疗组平均瘤重和瘤体积均较对照组明显减低,(P<0.01)。HIF-1α及VEGF在裸鼠肿瘤组织中均有阳性表达,其中对照组较AS2O3治疗组高(P<0.01)。结论:As2O3能够抑制体内外人结肠癌细胞的生长,并能明显抑制HIF-1α和VEGF的表达,从而抑制肿瘤的生长。

Objective： To discuss the mechanism of action through the investigation about the influence of proliferation by arsenic trioxide in human colon carcinoma LS-174T cell in vivo and vitro. Methods： 1. experimentation grouping： cell cultivation planning in vitro and internal animal experiment planning： control group; As2O3 low-dose group; As203 high-dose group.2, surveying cell proliferation condition by MTT. 3. detecting cell apoptosis ratio by AO/EB double fluorescein stain. 4. examing the expression of HIF-1α and VEGF in colon carcinoma cell and athymic mouse tumor tissue. 5. measureing the weight and volume of the tumor tissue in each athymic mouse group. Results： The growth speed and proliferating capacity of colon carcinoma cell LS-174T after AS2O3 effect were lower than control group in vitro, then As2O3 high-dose group were obviously. Cell apoptosis ratio in As2O3 group was higher than control group. Positive expression of HIF-1α and VEGF in control group affected colon carcinoma cell was higher than As2O3 group, and the expression in low-dose group was higher than high-dose group, with both the significant difference. In the internal animal experiment, the average weight and volume of transplantation tumor tissue in As2O33 group was obviously lower than control group. Positive expression of HIF-1α and VEGF can be seen in athymic mouse＇s transplantation tumor tissue, and the expression in control group was higher than As2O3 group. Conclusion： As2O3 can inhibit the growth of colon carcinoma cell LS-174T, and obviously inhibit the expression of HIF-1α and VEGF, accordingly restrain the growth of colon carcinoma cell.