芪玉三龙方调节Wnt/β-catenin通路下游靶基因表达抑制荷瘤小鼠肺癌生长

Research on Qiyu Sanlong Formula inhibiting growth of tumor-bearing mice of lung cancer by regulating downstream target genes experession levels of Wnt/β-catenin signaling pathway

目的:探讨芪玉三龙方对荷肺癌小鼠的抑瘤效果和对Wnt/β-catenin信号通路下游靶基因c-myc、cyclin D1、Axin2表达的影响。方法:采用Lewis肺癌细胞株培养移植法建立肺癌荷瘤小鼠模型,随机分为模型组,芪玉三龙方低、中、高剂量组,化疗组,联合组,每组8只;造模第2天开始予以芪玉三龙方低、中、高剂量组小鼠分别按20.12、40.24、80.48g·kg-1·d-1灌胃给药,化疗组按照0.4m L顺铂腹腔注射给药,联合组以高剂量中药灌胃和顺铂腹腔注射联合给药,模型组按照等量0.9%氯化钠溶液给药,连续给药21d;称取瘤质量,计算抑瘤率;RT-q PCR检测肿瘤组织c-myc、cyclin D1、Axin2 m RNA的转录水平。结果:与模型组和低剂量组比较,芪玉三龙方中、高剂量组和化疗组c-myc m RNA表达显著降低(P<0.01)。与模型组比较,各用药组cyclin D1 m RNA表达显著降低(P<0.05,P<0.01);与低剂量组比较,中剂量组、化疗组表达显著降低(P<0.01)。与模型组比较,各用药组Axin2 m RNA表达显著降低(P<0.01);与低剂量组、化疗组比较,芪玉三龙方中、高剂量组表达显著降低(P<0.01)。结论:芪玉三龙方对肺癌移植瘤具有一定的抑制作用,其抑瘤机制可能与芪玉三龙方调节肺癌组织c-myc m RNA、cyclin D1 m RNA、Axin2 m RNA的转录水平有关。芪玉三龙方高剂量抑瘤效应较优,量效关系明显,其与化疗药合用具有协同效应。

Objective： To explore the antitumor effects of Qiyu Sanlong Formula on lung cancer mice. And the effects of TCM compounds Qiyu Sanlong Formula on the expression levels of the downstream target genes（c-myc, cyclin D1, Axin2） of Wnt/β-catenin signaling pathway were explored. Methods： Tumor-bearing mice model of lung cancer was established by using Lewis lung cancer cell line Then, mice were randomly divided into model group, chemotherapy group and Qiyu Sanlong Formula of high, middle, low dose group, and joint group, eight mice in each group. From the second day after molding, Qiyu Sanlong Formula of low, medium and high dose group were given 20.12, 40.24, 80.48 g·kg-1·d-1 by gavage respectively. Mice in chemotherapy group were given intraperitoneal injection of 0.4 m L cisplatin, mice in joint group were given high dose of TCM by gavage and intraperitoneal injection. Mice in model group were given the same amount of normal saline. These drugs were administrated for successive 21 days. The tumor mass and the tumor inhibition rate were calculated. The m RNA transcription levels of c-myc, cyclin D1, Axin2 in tumor tissues were detected by RT-q PCR. Results： Compared with model group and Qiyu Sanlong Formula of low dose group, Qiyu Sanlong Formula of middle, high dose group and chemotherapy group decreased significantly in c-myc m RNA（P〈0.01）. Compared with model group, treatment groups decreased significantly in cyclin D1 m RNA（P〈0.05, P〈0.01）. Compared with Qiyu Sanlong Formula of low dose group, middle dose group and chemotherapy group decreased significantly in cyclin D1 m RNA（P〈0.01）. Compared with model group, the treatment groups decreased significantly in Axin2 m RNA（P〈0.01）. Compared with low dose group and chemotherapy group, Qiyu Sanlong Formula of middle, high dose group decreased significantly in Axin2 m RNA（P〈0.01）. Conclusion： Qiyu Sanlong Formula has certain inhibitory effect on transplantable lung cancer, and the anti-tumor mechanism might be related to the effect of Qiyu Sanlong Formula on regulating m RNA transcriptional levels of c-myc, cyclin D1, and Axin2 in lung cancer tissues. The high dose of Qiyu Sanlong Formula has a better antitumor effect and clear dose-effect relationship, and the combination treatment of Qiyu Sanlong Formula and chemotheraputic has synergistic effects.