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目的:探讨中药复方芪玉三龙汤(Qinyu Sanlong Decoction)对Wnt(Wingless,Intl的合称)信号通路拮抗基因分泌型Frizzled相关蛋白(SFRP)-2的影响。方法:基于前期研究基础,采用LLC细胞培养移植法,构建小鼠移植肺癌模型,将荷瘤小鼠随机分为模型组,芪玉三龙汤组(低、中、高剂量组),化疗组,联合组,芪玉三龙汤组小鼠分别按20.12,40.24,80.48g·kg~(-1)·d(-1)灌胃给药,化疗组以0.4mL顺铂腹腔注射给药(1次/周),联合组以芪玉三龙汤80.48 g·kg~(-1)·d~(-1)灌胃和腹腔注射顺铂联合给药,模型组以等体积0.9%氯化钠溶液灌胃,1次/d,连续给药21d;称取瘤质量,计算抑瘤率;透射电镜观察肿瘤细胞超微结构;蛋白免疫印迹法(Western blot)检测肿瘤组织β-链蛋白(β-catenin),SFRP-2蛋白表达。结果:与模型组相比,各用药组均可下调β-catenin蛋内表达(P<0.01);与苗玉三龙汤20.12 g·kg~(-1)·d~(-1)组比较,其他各用药组亦可下调0-catenin蛋白表达(P<0.01);与化疗组比较,芪玉三龙汤40.24,80.48 g·kg~(-1)d~(-1)组、联合组下调较明显(P<0.05<0.01)。与模型组相比,芪玉三龙汤40.24,80.48 g·kg~(-1)·d~(-1)组均可促进SFRP-2蛋白表达(P<0.05,P<0.01);与芪玉三龙汤20.12g·kg~(-1)d~(-1)组比较,其他各用药组亦可上调SFRP-2蛋白表达水平(P<0.01);与化疗组比较,芪玉三龙汤40.24,80.48g·kg~(-1)·d~(-1)组均可促进SFRP-2蛋白表达(P<0.05,P<0.01)。用药组电镜下可见凋亡现象,以芪玉三龙汤80.48g·kg~(-1)·d~(-1)组、化疗组及联合组明显。结论:芪玉三龙汤可一定程度抑制肺癌生长,诱导其镜下凋亡,促进SFRP-2表达,从而抑制Wnt/β-catenin信号通路。
OBJECTIVE: To investigate the effect of Qinyu Sanlong Decoction on the antagonist secreted Frizzled-related protein (SFRP) -2 of Wnt (the joint name of Wingless and Intl). Methods: Based on the previous research, LLC cell culture transplantation was used to establish a mouse lung cancer model. The tumor-bearing mice were randomly divided into model group, Qiyuandanlongtang group (low, middle and high dose), chemotherapy group , And the mice in the combination group and the stilbene group were treated by intragastric administration of 20.12, 40.24 and 80.48 g · kg -1 · d -1, respectively. The mice in the chemotherapy group were intraperitoneally injected with 0.4 mL cisplatin Once a week). The combination group was administered with 80.48 g · kg -1 · d -1 of stilbene trioxide and intraperitoneal injection of cisplatin. The model group was treated with 0.9% chlorinated The tumor volume was calculated and the tumor inhibition rate was calculated. The ultrastructure of tumor cells was observed by transmission electron microscopy. The expression of β-catenin in tumor tissue was detected by Western blot β-catenin), SFRP-2 protein expression. Results: Compared with the model group, the expression of β-catenin in each group was decreased (P <0.01), compared with 20.12 g · kg -1 · d -1 group (P <0.01). Compared with the chemotherapy group, Qiyu San Long Decoction 40.24 and 80.48 g · kg -1 d -1 group, the combination group Down significantly (P <0.05 <0.01). Compared with model group, SFQ-2 protein expression was promoted in 40.24,80.48 g · kg -1 · d -1 group (P <0.05, P <0.01) Compared with the chemotherapy group, the expression of SFRP-2 protein was also up-regulated in the other groups (20.12g · kg -1 d -1) (P <0.01) Soup 40.24, 80.48g · kg -1 d -1 could promote the expression of SFRP-2 protein (P <0.05, P <0.01). Apoptosis was observed under the electron microscope in the treatment group, which was obvious in the chemotherapy group and the combined group with 80.48g · kg -1 (-1) group. Conclusion: SJT can inhibit the growth of lung cancer to a certain extent, induce the apoptosis of the lung cancer, promote the expression of SFRP-2 and inhibit the Wnt / β-catenin signaling pathway.