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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 effect of esophagus Tongjie recipe on the proliferation and apoptosis of esophageal cancer cell line TE-1 in vivo to further clarify the mechanism of Esophageal cancer treatment. Methods The nude mice were inoculated subcutaneously with esophageal cancer TE-1 tumor strain. The nude mice bearing tumor were randomly divided into model group, TCM group, Cisplatin group and TCM + Cisplatin group, with 10 rats in each group. After 2 weeks in each group, the nude mice were sacrificed and the tumor mass was separated. The tumor weight was weighed and the tumor inhibition rate was calculated. The morphological changes and apoptosis of TE-1 cells were observed by HE staining. B-cell lymphoma-2 (Bcl-2) -2), cysteinyl aspartate proteinase (caspase 3) protein expression changes. Results The average tumor weight of the traditional Chinese medicine group, the cisplatin group and the traditional Chinese medicine + cisplatin group were lower than that of the model group (P <0.01), and the tumor inhibition rates were 45.95%, 48.65% and 56.76% The mean tumor weight was not significantly different from that of traditional Chinese medicine group (P> 0.05). The mean tumor weight of traditional Chinese medicine + cisplatin group was lower than that of traditional Chinese medicine group (P <0.01). In morphological changes, the morphology of the cells in the traditional Chinese medicine group, the cisplatin group and the traditional Chinese medicine + cisplatin group was round or oval, and the proportion of the cytoplasm to cytoplasm was decreased. The most significant change was in the traditional Chinese medicine plus cisplatin group. The results of immunohistochemistry showed that, The OD value of Bcl-2 in Chinese medicine group, cisplatin group and traditional Chinese medicine + cisplatin group was significantly lower than that in model group, with the lowest in Chinese traditional medicine + cisplatin group (P <0.01); compared with cisplatin group , The difference was not statistically significant (P> 0.05). The OD value of Bcl-2 in traditional Chinese medicine group was higher than that in traditional Chinese medicine + cisplatin group (P <0.01). The OD values of caspase 3 in Chinese medicine group, cisplatin group and traditional Chinese medicine + cisplatin group were significantly higher than those in model group, with the highest in Chinese traditional medicine + cisplatin group (P <0.01) (P> 0.05). The OD value of caspase 3 in traditional Chinese medicine group was lower than that in traditional Chinese medicine + cisplatin group (P <0.01). Conclusion Esophageal Tonglian Recipe can inhibit the proliferation and promote the apoptosis of esophageal cancer cell line TE-1, which may be related to down-regulating the expression of Bcl-2 and up-regulating the expression of caspase3 in esophageal cancer cell line TE-1.