中药复方WCAP及拆方对人胰腺癌皮下移植瘤裸小鼠模型及IL⁃6/STAT3信号通路的影响

Effects of traditional Chinese medicine compound formula WCAP and its sub⁃formulas on subcutaneous transplanted tumor nude mouse model of human pancreatic cancer and IL⁃6/STAT3 signaling pathway

目的观察中药复方WCAP及其拆方对人胰腺癌BxPC-3细胞裸小鼠皮下移植瘤生长及对白介素-6/信号转导和转录激活因子3(IL-6/STAT3)信号通路的影响。方法建立人胰腺癌BxPC-3细胞裸小鼠皮下移植瘤模型,将30只SPF级裸小鼠随机分为6组,即空白对照组、5-氟尿嘧啶(5-FU)组、复方组、健脾组、清热解毒组和软坚散结组,每组5只,观察各组裸小鼠皮下移植瘤瘤质量及抑瘤率,实时荧光定量逆转录聚合酶链式反应(RT-qPCR)和Western blot法分别检测瘤组织中IL-6/STAT3通路及其下游靶基因c-myc癌基因(c-myc)、细胞周期蛋白D1(cyclin D1)、血管内皮生长因子(VEGF)、基质金属蛋白酶-2(MMP-2)的mRNA和蛋白表达水平。结果①与空白对照组比较,各干预组瘤质量均轻于空白对照组,5-FU组、复方组及各拆方组均可抑制裸小鼠皮下移植瘤的生长(P<0.05),复方组抑瘤率高于各拆方组(P<0.05)。②与空白对照组比较,5-FU组、复方组和软坚散结组IL-6/STAT3通路及其下游靶基因c-myc、cyclin D1、VEGF、MMP-2的mRNA表达均降低(P<0.05)。③与空白对照组比较,5-FU组、复方组IL-6/STAT3通路及其下游靶基因涉及的c-myc、cyclin D1、VEGF、MMP-2蛋白表达均降低(P<0.05),软坚散结组c-myc、MMP-2和IL-6蛋白表达均降低(P<0.05)。结论WCAP复方可抑制人胰腺癌BxPC-3细胞裸小鼠皮下移植瘤的生长,其机制与抑制IL-6/STAT3通路并调节其下游靶基因c-myc、cyclin D1、VEGF、MMP-2的表达水平有关;与各拆方相比,WCAP复方在抑瘤率、小鼠体质量及IL-6/STAT3通路及其下游靶基因调控方面作用更好。

Objective To study the effects of traditional Chinese medicine(TCM)compound formula WCAP and its sub-formulas on the tumor growth of subcutaneous transplanted human pancreatic cancer BxPC-3 cells in nude mice and on the interleukin-6/signal transducer and activator transcription 3(IL-6/STAT3)signaling pathway.Methods The subcutaneous transplanted tumor model of human pancreatic cancer BxPC-3 cells in nude mice was established.Totally 30 SPF-grade nude mice were randomly divided into 6 groups of 5 mice each,which were blank control group,5-fluorouracil(5-FU)group,compound group,spleen-invigorating group(SI group),heat-clearing and toxicity-removing group(HCTR group),and hardness-resolving group(HR group).The tumor quality and tumor growth inhibition rate of the subcutaneous transplanted tumor in each group of nude mice were recorded.The mRNA and protein expression levels of IL-6/STAT3 pathway and its downstream target genes c-myc,cyclin D1,VEGF and MMP-2 were detected in tumor tissues by real-time fluorescence quantitative reverse transcription polymerase chain reaction(RT-qPCR)and Western blot,respectively.Results①Each intervention group showed lower tumor weight than that of control group.The 5-FU group,compound group and each sub-formula group could inhibit the growth of subcutaneous transplanted tumor in nude mice(P<0.05),and the tumor growth inhibition rate of the compound group was higher than that of each sub-formula group(P<0.05).②The mRNA expression levels of IL-6/STAT3 pathway and its downstream target genes c-myc,cyclin D1,VEGF and MMP-2 were lowered in the 5-FU group,compound group and HR group than those in the control group(P<0.05).③Compared with those in the control group,the protein expression levels of IL-6/STAT3 pathway and its downstream target genes c-myc,cyclin D1,VEGF,and MMP-2 were lowered in the 5-FU group and compound group(P<0.05),and the protein expression levels of c-myc,MMP-2,and IL-6 were lowered in HR group(P<0.05).Conclusions WCAP compound formula could inhibit the tumor growth of subcutaneous transplanted human pancreatic cancer BxPC-3 cells in nude mice,and the mechanism was related to the inhibition of IL-6/STAT3 pathway and regulation of the expression levels of its downstream target genes c-myc,cyclin D1,VEGF,and MMP-2.WCAP compound formula showed a better effect on the tumor growth inhibition rate,mouse weight,and IL-6/STAT3 pathway and its downstream target gene regulation than each sub-formula group.