柴胡龙牡汤联合卡培他滨对三阴性乳腺癌荷瘤裸鼠抑瘤作用及具体机制

Antitumor effect of chaihu longmu prescription combined with capecitabine on triple negative breast cancer mice and its mechanism

目的探讨柴胡龙牡汤联合卡培他滨抑制三阴性乳腺癌(TNBC)荷瘤裸鼠肿瘤生长的影响,并阐明在转化生长因子(TGF)通路中可能的作用机制。方法40只雌性BALB/c-nu裸鼠随机分为5组:空白对照组、模型对照组、柴胡龙牡汤组、卡培他滨组及联合组(柴胡龙牡汤+卡培他滨),每组8只,于裸鼠胸骨前乳垫处接种MDA-MB-231细胞,构建TNBC模型。灌胃给药按人鼠体表面积换算后,模型组等剂量生理盐水灌胃,灌胃周期21 d。结束取出各组瘤体称重,计算抑瘤率。HE染色、马松染色检测肿瘤组织病理形态学改变,实时荧光定量PCR(RT-qPCR)检测各组肿瘤组织中TGF-β1 mRNA、Smad3 mRNA、Smad7 mRNA表达水平。结果与模型组相比,柴胡龙牡汤组、卡培他滨组及联合组裸鼠肿瘤组织质量均有所下降,其中联合组下降最明显(F=85.710,P=0.002)。柴胡龙牡汤组、卡培他滨组及联合组裸鼠肿瘤组织凋亡区域明显增加,肿瘤组织纤维化程度明显降低。与模型组相比,联合组、卡培他滨组、柴胡龙牡汤组裸鼠肿瘤组织中TGF-β1 mRNA、Smad3 mRNA表达水平均降低,Smad7 mRNA表达均增加(F=43.210,P<0.001;F=49.240,P<0.001;F=20.350,P<0.001)。其中联合组中TGF-β1 mRNA、Smad3 mRNA基因表达下调最明显,Smad7 mRNA表达上调最明显。结论柴胡龙牡汤联合卡培他滨有抑制TNBC荷瘤裸鼠肿瘤生长的作用,其可能通过降低TGF-β1 mRNA、Smad3 mRNA表达,升高Smad7 mRNA表达促进肿瘤细胞凋亡,并可能通过调控TGF信号通路来干预TNBC的疾病进展。

Objective To investigate the effect of chaihu longmu prescription combined with capecitabine on tumor growth in triple-negative breast cancer(TNBC)mice,and to clarify the possible mechanism of transforming growth factor(TGF)pathway.Methods Forty female BALB/c-nu nude mice were randomly divided into five groups:the blank control group,the model control group,the chaihu longmu prescription group,the capecitabine group and the combined group(chaihu longmu prescription+capecitabine),8 mice in every group.TNBC model was constructed by inoculating MDA-MB-231 cells in the breast pad of nude mice.Gavage was administered according to the body surface area of rats and humans.The model group was given equal dose of normal saline.The gavage cycle was 21 d.Finally,the tumor body of each group was taken out and weighed to calculate the tumor inhibition rate.HE staining and Masson staining were used to detect the pathological changes of tumor tissue,and the expressions of TGF-β1 mRNA,Smad3 mRNA and Smad7 mRNA in tumor tissues of each group were detected by real-time fluorescence quantitative PCR(RT-qPCR).Results Compared with the model group,the weight of tumor tissue in the capecitabine group,the chaihu longmu prescription group and the combination group were decreased,and the weight of tumor tissue in the combination group was significantly decreased(F=85.710,P=0.002).The apoptotic area of tumor tissue increased significantly and the degree of fibrosis of tumor tissue decreased significantly in the capecitabine group,the chaihu longmu prescription group and the combination group.Compared with the model group,the expression of TGF-β1 mRNA and Smad3 mRNA in nude mice in the capecitabine group,the chaihu longmu prescription group and the combination group were decreased,while the expression of Smad7 mRNA was increased(F=43.210,P<0.001;F=49.240,P<0.001;F=20.350,P<0.001).In the combination group,TGF-β1 mRNA and Smad3mRNA were most significantly down-regulated,while Smad7 mRNA was most significantly up-regulated.Conclusion Combined with capecitabine,chaihu longmu prescription can inhibit the tumor growth of TNBC in nude mice with tumor bearing tumor.The mechanism may reduce the expression of TGF-β1 mRNA and SMad3 mRNA,increase the expression of SMad7 mRNA and promote the apoptosis of tumor cells,and may affect TNBC by regulating the TGF signaling pathway.