健脾解毒方对大肠癌细胞COX-2/β-catenin/MMP-7信号通路的影响  

Effect of Jianpi Jiedu Recipe on colorectal cancer cells via COX-2/β-catenin/MMP-7signal pathway

作　　者：刘宣[1] 周利红[1] 隋华[1] 柴妮[1] 江海丽 陈静[1] 孙筱婷 李琦[1] 

机构地区：[1]上海中医药大学附属曙光医院肿瘤科,上海201203

出　　处：《上海中医药杂志》2016年第4期65-69,共5页Shanghai Journal of Traditional Chinese Medicine

基　　金：国家自然科学基金资助项目(81303103;81473478);上海市科委科研基金资助项目(13ZR1461000;14140901402);上海市教委"晨光计划"资助项目(13CG47)

摘　　要：目的探讨健脾解毒方对大肠癌细胞转移能力的影响及可能的作用机制。方法采用CCK-8检测不同浓度健脾解毒方醇提物对Lo Vo细胞生长的抑制作用。分别采用健脾解毒方、过表达环氧合酶-2(COX-2)基因和(或)糖原合成激酶-3β(GSK-3β)选择性抑制剂SB-216763作用Lo Vo细胞24h,Transwell检测细胞转移能力;Western blottting检测细胞中COX-2和β-连环蛋白(β-catenin)的蛋白表达;ELISA检测细胞培养上清液中基质金属蛋白酶-7(MMP-7)表达。结果健脾解毒方能够以剂量和时间依赖性方式抑制Lo Vo细胞的生长。与空白对照组比较,健脾解毒方能够明显抑制Lo Vo细胞的转移能力(P<0.01),明显下调Lo Vo细胞中COX-2蛋白表达,下调β-catenin在细胞核内的积累及其下游靶基因MMP-7的表达(P<0.01)。COX-2基因能够增加β-catenin在细胞核内的积累及其MMP-7的表达,促进Lo Vo细胞转移(P<0.01);而上调COX-2表达能够逆转健脾解毒方对Lo Vo细胞中β-catenin在细胞核中的积累及其MMP-7的表达和转移的抑制作用,激活β-catenin信号通路能够逆转健脾解毒方对Lo Vo细胞转移的抑制作用。结论健脾解毒方能够抑制人肠癌细胞的转移能力,其作用机制可能与抑制COX-2/β-catenin/MMP-7信号通路有关。Objective To explore the effects and possible mechanism of Jian Pi Jie Du Recipe（ JPJD） on metastasis of colorectal cancer. Methods The effect of JPJD on the proliferation of colorectal cancer Lo Vo cells was tested by CCK-8 assay. Using JPJD,over-expression of COX-2 Gene and / or GSK-3β selective inhibitor SB-216763 in Lo Vo cells for 24 h,the migration of Lo Vo cells was analyzed by Transwell assay; the expression of COX-2 and β-catenin protein was detected by western blot; MMP-7 expression in cell culture supernatant was assayed by ELISA. Results JPJD alcohol extracts had obvious inhibition on the proliferation of Lo Vo cells in dose and time-dependent manner. JPJD could down-regulate the migratory ability of Lo Vo cells compared with the control group（ P 0. 01）. JPJD could down-regulated the expression of COX-2,reduced the accumulation of β-catenin in the nuclear of Lo Vo cells,following by the decrease of the β-catenin targeted MMP-7 expression（ P 0. 01）. COX-2 could increase the accumulation of β-catenin in the nuclear and promote the expression of β-catenin targeted MMP-7,leading to increased metastatic potential of Lo Vo cells（ P 0. 01）. We further showed that an increase of COX-2 expression offset the repressive activity of JPJD on β-catenin and MMP-7 expression; activating of β-catenin signal pathway rescued Lo Vo cells from suppressed migration by JPJD.Conclusion Jian Pi Jie Du Recipe inhibited CRC metastasis may via down-regulation of COX-2 / β-catenin / MMP-7 signaling pathway.

关 键 词：健脾解毒方 大肠癌 环氧合酶-2 Β-连环蛋白 基质金属蛋白酶-7 体外实验 

分 类 号：R285[医药卫生—中药学]