【作者】 刘宣； 陈静； 孙筱婷； 李琦；

【Author】 LIU Xuan;CHEN Jing;SUN Xiaoting;LI Qi;Department of Medical Oncology,Shuguang Hospital,Shanghai University of Traditional Chinese Medicine;

【机构】 上海中医药大学附属曙光医院肿瘤科；

【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机制。方法:采用CCK-8检测不同浓度健脾解毒方醇提物对LoVo细胞生长的抑制作用。分别采用健脾解毒方、过表达COX-2基因和/或GSK-3β选择性抑制剂SB-216763作用LoVo细胞24h,Transwell检测细胞转移能力;Western blot检测细胞中COX-2和β-catenin蛋白表达;ELISA检测细胞培养上清液中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LoVo细胞的生长。与对照组比较,健脾解毒方能够明显抑制LoVo细胞的转移能力(P<0.01)。与对照组比较,健脾解毒方能够明显下调LoVo细胞中COX-2蛋白表达,以及下调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LoVo细胞转移(P<0.01)。上调COX-2表达能够逆转健脾解毒方对LoVo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LoVo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。更多还原

【Abstract】 Objective To explore the effects and possible mechanism of JianPi JieDu Recipe(JPJD) on metastasis of colorectal cancer.Methods The effect of JPJD on the proliferation of colorectal cancer LoVo cells was tested by CCK-8 assay.Using JPJD,over-expression of COX-2 Gene and/or GSK-3β selective inhibitor SB-216763 in LoVo cells for 24 h,the migration of LoVo 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 LoVo cells in dose and timedependent manner.JPJD could down-regulate the migratory ability of LoVo cells compared with the control group(P<0.01).JPJD could down-regulated the expression of COX-2,reduced the accumulation of P-catenin in the nuclear of LoVo cells,following by the decrease of the p-catenin targeted MMP-7 expression(P<0.01).COX-2 could increase the accumulation of P-catenin in the nuclear and promote the expression of β-catenin targeted MMP-7,leading to increased metastatic potential of LoVo 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 LoVo cells from suppressed migration by JPJD.Conclusion JPJD inhibited CRC metastasis via down-regulation of COX-2/β-catenin/MMP-7 signaling pathway.更多还原