半枝莲抑制结肠癌细胞迁移和侵袭实验研究

Experimental study on inhibition of migration and invasion of colon cancer cells by scutellaria barbata

目的:探讨半枝莲对结肠癌细胞迁移和侵袭的抑制作用及相关机制。方法:体外培养结肠癌SW620、CCL-228细胞系,用浓度为10μmol/L半枝莲进行干预。将SW620、CCL-228细胞分为对照组(不做任何处理)、10μmol/L半枝莲组、Rac1L61组(Rac1L61过表达质粒转染细胞)、联合组(Rac1L61过表达质粒转染细胞后加入10μmol/L半枝莲)。实时细胞分析(RTCA)系统评估半枝莲对细胞的抑制作用,MTT法检测细胞活力,划痕实验检测细胞迁移,Transwell实验检测细胞侵袭,Western blotting法检测Rac1、p-PAK1、MMP2、MMP9蛋白表达水平。结果:10μmol/L半枝莲组SW620、CCL-228细胞中Rac1、p-PAK1蛋白表达均低于对照组,差异有统计学意义(均P<0.05)。转染过表达Rac1质粒后,SW620、CCL-228细胞Rac1、p-PAK1蛋白表达均升高,而用10μmol/L半枝莲干预可逆转这种效应。10μmol/L半枝莲组SW620、CCL-228细胞侵袭和迁移能力均低于对照组,差异有统计学意义(均P<0.05)。转染过表达Rac1质粒后,SW620、CCL-228细胞侵袭和迁移明显增强,而用10μmol/L半枝莲干预可逆转这种效应。10μmol/L半枝莲组MMP2、MMP9蛋白表达低于对照组(均P<0.05)。转染过表达Rac1质粒后,SW620、CCL-228细胞MMP2、MMP9蛋白表达均升高,而用10μmol/L半枝莲干预可逆转这种效应。结论:半枝莲通过调节Rac1/PAK1信号通路活性,减少MMP9、MMP2表达而降低结肠癌细胞迁移和侵袭能力。

Objective:To investigate inhibitory effect of scutellaria barbata on migration and invasion of colon cancer cells and its mechanism.Methods:Colon cancer SW620,CCL-228 cell lines were cultured in vitro with 10μmol/L scutellaria barbata was intervened.SW620 and CCL-228 cells were divided into control group(without any treatment),10μmol/L scutellaria barbata group(add 10μmol/L scutellaria barbata),Rac1L61 group(transfected with Rac1L61 overexpression plasmid),combine group(after transfection with Rac1L61 overexpression plasmid add 10μmol/L scutellaria barbata).Real time cell analysis(RTCA)system was used to evaluate inhibitory effect of scutellaria barbata on cells,cell viability detected by MTT assay,cell migration detected by scratch test,transwell assay used to detect cell invasion.The protein expression levels of Rac1,p-PAK1,MMP2 and MMP9 were detected by Western blotting.Results:Expression of Rac1 and p-PAK1 protein in SW620 and CCL-228 cells of 10μmol/L scutellaria barbata group were lower than those in the control group,difference statistically significant(all P<0.05).After transfection with Rac1 plasmid,the expression levels of Rac1 and p-PAK1 protein in SW620 and CCL-228 cells were significantly increased,while the expression levels of 10μmol/L scutellaria barbata intervention cells can reverse this effect.The invasion and migration of SW620 and CCL-228 cells in 10μmol/L scutellaria barbata group were lower than those in the control group,difference statistically significant(all P<0.05).After transfection with Rac1 plasmid,the invasion and migration of SW620 and CCL-228 cells were significantly enhanced,but 10μmol/L scutellaria barbata intervention cells can reverse this effect.The expression level of MMP2 and MMP9 protein in 10μmol/L scutellaria barbata group were lower than those in the control group(all P<0.05).After transfection with Rac1 plasmid,the expression level of MMP2 and MMP9 protein in SW620 and CCL-228 cells increased,while the expression level of 10μmol/L scutellaria barbata intervention cells can reverse this effect.Conclusion:Scutellaria barbata can reduce the ability of migration and invasion of colon cancer cells by regulating the activity of Rac1/PAK1 signal pathway and reducing the expression of MMP9 and MMP2.