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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 investigate the effect of Jianpi Jiedu Formula on the metastasis of colorectal cancer cells and its possible mechanism. Methods CCK-8 was used to detect the inhibitory effect of different concentrations of Jianpi Jiedu Fang alcohol extract on LoVo cell growth. Transfection of LoVo cells was induced by Jianpi Jiedu decoction, COX-2 gene overexpression and SB-216763, a selective inhibitor of glycogen synthesis kinase-3β (GSK-3β) The protein expression of COX-2 and β-catenin in the cells was detected by Western blottting. The expression of MMP-7 in the cell culture supernatant was detected by ELISA. Results Jianpijiedu Decoction could inhibit the growth of LoVo cells in a dose-and time-dependent manner. Compared with the blank control group, Jianpi Jiedu Decoction could significantly inhibit the metastasis ability of LoVo cells (P <0.01), down-regulate the expression of COX-2 protein and down-regulate the accumulation of beta-catenin in LoVo cells and its downstream target Gene MMP-7 expression (P <0.01). COX-2 gene could increase the accumulation of β-catenin in the nucleus and the expression of MMP-7, and promote the metastasis of LoVo cells (P <0.01). Up-regulation of COX-2 could reverse the effect of Jianpi Jiedu Decoction on LoVo cells β-catenin accumulation in the nucleus and the inhibition of MMP-7 expression and metastasis, and activation of β-catenin signaling pathway can reverse the inhibition of Jianpijieduifu on LoVo cell metastasis. Conclusion Jianpi Jiedu Decoction can inhibit the metastasis of human colon cancer cells, and its mechanism may be related to the inhibition of COX-2 / β-catenin / MMP-7 signaling pathway.