解毒祛瘀方对MCF-7人乳腺癌细胞侵袭能力的影响及机制探讨

Role and Mechanism of Jiedu Quyu Decoction on Invasion of Breast Cancer Cell

目的:研究解毒祛瘀方对人乳腺癌细胞侵袭能力的影响及机制。方法:以人乳腺癌MCF-7细胞为研究对象,预培养后分为空白组,解毒祛瘀方低、中、高剂量组(0.625,1.25,2.50 g·L-1),阳性药组(5-氟尿嘧啶,10 mg·L-1),分别用0.625,1.25,2.50 g·L-1的解毒祛瘀方及5-氟尿嘧啶处理人乳腺癌MCF-7细胞24 h后,MTT法检测细胞黏附率,Transwell小室实验检测细胞侵袭能力,RT-PCR法检测肿瘤细胞CD44,E-钙黏蛋白及N-钙黏蛋白基因的表达,Western blot法检测肿瘤细胞CD44,E-钙黏蛋白,N-钙黏蛋白,磷酸化细胞外信号调节激酶(p-ERK)及磷酸化蛋白激酶B(p-Akt)蛋白的表达。结果:与空白组比较,解毒祛瘀方低、中、高剂量组可明显抑制人乳腺癌MCF-7细胞的黏附和侵袭能力;且使CD44及E-钙黏蛋白基因及蛋白表达上调,N-钙黏蛋白表达下调,并可以抑制PI3K/Akt及MEK/ERK信号通路的激活(P<0.05,P<0.01)。结论:解毒祛瘀方可抑制人乳腺癌MCF-7细胞体外侵袭能力,其机制可能与抑制相关异质黏附蛋白的表达、抑制上皮间质转化及调节肿瘤相关信号通路有关。

Objective： To research Jiedu Quyu decoction effects on human breast cancer cell of invasion ability and mechanism. Method： Respectively,0. 625,1. 25,2. 50 g·L- 1Jiedu Quyu decoction were treated with human breast cancer MCF-7 cell,adhesion assay and transwell assay was performed to test the adhesion ability and invasion ability. RT-PCR and Western blot were used to analyze the expression changes of genes and proteins,including E-cadherin,N-cadherin,CD44,extracellular signal-regulated kinase（ ERK）,protein kinase B（ Akt）.Result： Compared with the blank group,treated with 0. 625,1. 25,2. 50 g·L- 1Jiedu Quyu decoction,the adhesion and invasion ability of MCF-7 can be inhibited obviously. The expression of CD44 and E-cadherin were up-regulated,while the N-cadherin were down-regulated. And the signaling pathway of PI3 K / Akt and MEK / ERK were inhibited（ P〈0. 05,P〈0. 01）. Conclusion： Jiedu Quyu decoction could inhibit the invasion of MCF-7. Its mechanism may be correlated with inhibition of heterogeneous adhesion protein expression,and inhibition of epithelial-mesenchymal transition and regulation of signaling pathway.