食道通结方对裸鼠食管癌移植瘤中TE-1细胞增殖及凋亡的影响

Influence of “Shidao Tongjie Decoction”on the growth and apoptosis of TE-1 cells in nude mice transplanted with esophagus cancer

目的通过体内实验观察食道通结方对食管癌细胞株TE-1细胞增殖和凋亡的影响,以进一步明确该方治疗食管癌的作用机制。方法裸鼠皮下接种食管癌TE-1瘤株,将荷瘤裸鼠随机分成模型组、中药组、顺铂组、中药+顺铂组,每组10只,其中中药选取食道通结方。各组均灌胃2周后,处死裸鼠,分离瘤块。称取瘤重,计算肿瘤抑制率;HE染色观察TE-1细胞形态的变化、细胞凋亡的变化;免疫组化检测肿瘤组织内B淋巴细胞瘤-2基因(B-cell lymphoma-2,Bcl-2)、含半胱氨酸的天冬氨酸蛋白水解酶(cysteinyl aspartate specific proteinase 3,caspase 3)蛋白表达的变化。结果中药组、顺铂组和中药+顺铂组平均瘤重均低于模型组,差异有统计学意义(P<0.01),抑瘤率分别为45.95%、48.65%、56.76%;顺铂组平均瘤重与中药组相比差异无统计学意义(P>0.05),中药+顺铂组平均瘤重低于中药组,差异有统计学意义(P<0.01)。在形态变化上,中药组、顺铂组和中药+顺铂组细胞部分形态呈圆形或卵圆形,细胞核浆比例减小,其中中药+顺铂组变化最为显著;免疫组化结果显示,中药组、顺铂组和中药+顺铂组Bcl-2 OD值明显低于模型组,其中以中药+顺铂组最低,差异有统计学意义(P<0.01);中药组与顺铂组比较,差异无统计学意义(P>0.05),中药组Bcl-2 OD值高于中药+顺铂组,差异有统计学意义(P<0.01)。中药组、顺铂组和中药+顺铂组caspase 3的OD值明显高于模型组,其中以中药+顺铂组最高,差异有统计学意义(P<0.01);中药组与顺铂组相比差异无统计学意义(P>0.05),中药组caspase 3的OD值低于中药+顺铂组,差异有统计学意义(P<0.01)。结论食道通结方具有抑制食管癌细胞株TE-1增殖、促进其凋亡的作用,该作用可能与下调食管癌细胞株TE-1中Bcl-2的表达,上调caspase3的表达相关。

Objective To observe the effects of ＂Shidao Tongjie Decoction＂ （SDTJ） on cell proliferation and apoptosis of esophagus cancer cell line （ TE- 1 ） in vivo, and further investigate the mechanism of action of SDTJ in the treatment of esophagus cancer. Methods TE- 1 cells were inoculated subcutaneously into the nude mice. The nude mice bearing tumor were then randomly divided into the following 4 groups ： model group, SDTJ group, cisplatin group and combination group （SDTJ combined with cisplatin）, with 10 mice in each group： The mice in each group were administrated by gastric infusion for 2 weeks. Then the mice were executed, the tumor weight and the inhibition rate were calculated. The change of TE-1 cells morphology and apoptosis were observed by HE staining; Protein expressions of Bcl-2 and caspase3 were detected by immunohistochemical method. Results The tumor weight of the three treatment groups was all lower than that of the model group （P 〈 0.01 ）, the tumor inhibition rate was 49.95 %, 48.65 % and 56.76% respectively; there＇ s no statistical significance （P = 0. 346 ） in tumor weight between SDTJ group and cisplatin group; the tumor weight of the combination group was lower than SDTJ group（ P 〈0.01 ）. In the form changing aspect, the SDTJ group and combination group had the lower cell density, the shape of the cells was roundness or elliptical, the atypical epithelial hyperplasia hepatolithiasis was reduced, the combination group had the most prominent change. Immunohistochemical result showed that the expression of Bcl-2 in SDTJ group, cisplatin group and combination group was obviously lower than that in the model group, with the combination group having the lowest expression （P 〈 0.01 ）, and with no statistical significance （P = 0. 581 ） between SDTJ group and cisplatin group. The caspase 3 expression in SDTJ group, cisplatin group and combination group was obviously higher than that in model group, with combination group having the highest expression （ P 〈0.01 ）, and with no significant difference between SDTJ group and cisplatin group. Conclusion Shi Dao Tong Jie formula could restrain the proliferation and promote the apoptosis of TE- 1 esophagus cancer cell line, which may be related to the down-regulation of the Bcl-2 expression and up-regulation of the caspase 3 expression in the esophagus cancer cell line.