黄芪—莪术—重楼配伍对结肠癌原位移植瘤模型裸鼠肿瘤及癌旁组织中侵袭性伪足相关蛋白表达的影响

Effect of Huangqi-Ezhu-Chonglou(黄苗-莪术-重楼) Combination on the Expression of Invadopodia-related Proteins in the Tumors and Paracancerous Tissues of Nude Mice with Colon Cancer-derived Orthotopic Transplant Model

目的探讨黄芪-莪术-重楼配伍抗结肠癌转移的可能作用机制。方法38只裸鼠采用随机数字表法分为空白组8只及模型组、5-氟尿嘧啶(5-Fu)组和黄芪-莪术-重楼组各10只。除空白组外,其余各组裸鼠采用人结肠癌HCT116细胞原位移植法构建结肠癌原位移植瘤模型。造模24h后,5-Fu组给予注射用5-Fu 25mg/(kg·d)腹腔注射,隔日1次;黄芪-莪术-重楼组给予5.85g/(kg·d)黄芪-莪术-重楼药液灌胃,每日1次;空白组、模型组分别以10ml/(kg·d)生理盐水灌胃,每日1次。连续给药21天后计算瘤重及抑瘤率;检测各组裸鼠肿瘤及癌旁组织中酪氨酸激酶底物5(Tks5)、皮层肌动蛋白(Cortactin)、磷酸化皮层肌动蛋白(p-Cortactin)、细胞分裂周期蛋白(Cdc42)、衔接蛋白1(Nck1)、丝状肌动蛋白(F-actin)及膜型基质金属蛋白酶1(MT1-MMP)蛋白表达。结果5-Fu组、黄芪-莪术-重楼组的抑瘤率分别为58.32%、45.77%。与模型组比较,黄芪-莪术-重楼组、5-Fu组平均瘤重均降低(P<0.01);5-Fu组与黄芪-莪术-重楼组平均瘤重比较差异无统计学意义(P>0.05)。与空白组比较,模型组、黄芪-莪术-重楼组及5-Fu组肿瘤组织及癌旁组织中侵袭性伪足前体核心蛋白Tks5、p-Cortactin,侵袭性伪足成熟相关蛋白Cdc42、Nck1、F-actin、MT1-MMP蛋白表达均升高(P<0.05或P<0.01)。与模型组比较,黄芪-莪术-重楼组及5-Fu组肿瘤组织及癌旁组织中侵袭性伪足前体核心蛋白Tks5、p-Cortactin,侵袭性伪足成熟相关蛋白Cdc42、Nck1、F-actin、MT1-MMP蛋白表达均降低(P<0.01)。结论黄芪-莪术-重楼配伍可通过抑制肿瘤侵袭性伪足的形成及成熟,从而抑制结肠癌的生长与转移。

Objective To explore the possible mechanism of Huangqi-Ezhu-Chonglou(黄芪-莪术-重楼,Qi-Zhu-Lou)combination on metastatic colon cancer.Methods Thirty-eight nude mice were divided into blank group(n=8),model group(n=10),5-fluorouracil(5-Fu)group(n=10)and Qi-Zhu-Lou group(n=10)by using a random number table.The nude mice in all groups but the blank group were transplanted with human colon cancer HCT116 cells following the orthotopic transplantation method to develop the colon cancer-derived orthotopic transplant model.Twenty-four hours after modeling,the 5-Fu group was given intraperitoneal injection of 25 mg/(kg·d)5-Fu,once every other day;the Qi-Zhu-Lou group was administered with Qi-Zhu-Lou liquid of 5.85 g/(kg·d)by gavage,once daily;both the blank group and the model group were given 10 ml/(kg·d)normal saline by gavage,once daily.After 21 days of continuous administration,all nude mice were killed to calculate tumor weight and tumor inhibition rate;the expression of tyrosine kinase substrate 5(Tks5),cortical actin(Cortactin),phosphorylated cortical actin(p-Cortactin),cell division cyclin(Cdc42),adaptor protein 1(Nck1),filamentous actin(F-actin)and membrane-type matrix metalloproteinase 1(MT1-MMP)protein in tumors and paracancerous tissues of nude mice was detected in each group.Results The tumor inhibition rate of the 5-Fu group and the Qi-Zhu-Lou group was 58.32% and 45.77%,respectively,significantly lower than that of the model group(P<0.01),and with no significant difference between the two groups(P>0.05).The protein expressions of invadopodia core protein precursor Tks5 and p-Cortactin,as well as invadopodia maturation-associated protein Cdc42,Nck1,F-actin and MT1-MMP in the tumor and paracancerous tissues of the mice in the model group,Qi-Zhu-Lou group and 5-FU group were significantly higher than those in the blank group(P<0.05 or P<0.01),while lower than those in the model group(P<0.01).Conclusion Qi-Zhu-Lou combination may inhibit the growth and metastasis of colon cancer cells by inhibiting the formation and maturation of invadopodia.