阳和化岩汤对HER-2高表达型乳腺癌细胞PI3K/Akt通路HER-2,PI3K,p-Akt表达的机制

Effect of Yanghe Huayan Tang Combined with PI3K Inhibitor on Expressions of HER-2,PI3K and p-Akt in PI3K/Akt Pathway of High-expression HER-2 Breast Cancer Cell Lines

目的:观察阳和化岩汤与磷脂酰肌醇-3羟基激酶(PI3K)抑制剂LY294002对乳腺癌细胞株SK-BR-3[人表皮生长因子受体-2(HER-2)高表达型]PI3K/蛋白激酶B(Akt)信号通路中HER-2,PI3K,磷酸化蛋白激酶B(p-Akt)表达的影响,研究阳和化岩汤的干预机制。方法:制备阳和化岩汤肠吸收液,将乳腺癌细胞株SK-BR-3分为空白组,阳和化岩汤组,LY294002组,LY294002+阳和化岩汤组,阳和化岩汤(125 g·L-1),LY294002 (0. 05 g·L-1),阳和化岩汤+LY294002 (阳和化岩汤125 g·L-1,LY294002 0. 05 g·L-1),用药干预24 h后通过免疫组化、蛋白免疫印迹法(Western blot)检测HER-2,PI3K,p-Akt蛋白表达,通过逆转录聚合酶链式反应(RT-PCR)检测HER-2,PI3K,p-Akt mRNA表达。结果:与空白组比较,LY294002组,LY294002+阳和化岩汤组均能抑制乳腺癌细胞株HER-2,PI3K,p-Akt蛋白及mRNA表达(P <0. 05);阳和化岩汤组能抑制乳腺癌细胞株HER-2,p-Akt蛋白及mRNA表达(P <0. 05),但对PI3K的抑制作用不明显;联用LY294002较单用LY294002具有进一步抑制PI3K表达的作用(P <0. 05)。与阳和化岩汤组,LY294002组比较,LY294002+阳和化岩汤组能够明显抑制HER-2,PI3K,p-Akt蛋白及mRNA表达(P <0. 05)。结论:阳和化岩汤能够抑制乳腺癌脉管生成和侵袭,主要作用机制为通过干预PI3K/Akt通路,降低通路中HER-2,PI3K,p-Akt表达。

Objective: To investigate effect of Yanghe Huayan Tang and phosphatidylinositol 3-kinase(PI3 K) inhibitor LY294002 in intervening expressions of human epidermal growth factor receptor human epidermal growth factor receptor-2(HER-2),PI3 K and phosphorylated serine/threonine kinase(p-Akt) in PI3 K/Akt pathway of breast cancer cell line SK-BR-3(HER-2 high expression) in vitro,and study intervention mechanism of Yanghe Huayan Tang. Method: Yanghe Huayan Tang intestinal absorption fluid was prepared,breast cancer cell strain SK-BR-3 was divided into blank group,Yanghe Huayan Tang group,LY294002 group,LY294002 + Yanghe Huayan Tang group. The concentration was respectively 125 g·L-1 for Yanghe Huayan Tang,0. 05 g·L-1 for LY294002,125 g·L-1 for Yanghe Huayan Tang + LY294002. The expressions of HER-2,PI3 K and p-Akt protein were detected by immunohistochemistry and Western blot after 24 h. The most appropriate and effective concentration was obtained through extensive experiments. The expressions of HER-2,PI3 K and p-Akt were detected by reverse transcription polymerase chain reaction(RT-PCR). Result: Compared with blank group,LY294002 group,and LY294002 + Yanghe Huayan Tang group could inhibit protein and mRNA expressions of HER-2,PI3 K, p-Akt in breast cancer cell lines(P < 0. 05). Yanghe Huayan Tang group could inhibit expressions of HER-2,p-Akt and mRNA(P < 0. 05) in breast cancer cell lines,but inhibitory effect on PI3 K was not obvious;combination with LY294002 could further inhibit PI3 K expression compared with single administration with LY294002(P < 0. 05). Compared with LY294002 group,LY294002 + Yanghe Huayan Tang group had a more obvious inhibitory effect. The combined administration of Yanghe Huayan Tang and LY294002 could inhibit protein and mRNA expressions of HER-2,PI3 K,p-Akt significantly(P < 0. 05). Conclusion: Yanghe Huayan Tang can inhibit angiogenesis and invasion of breast cancer. Its main mechanism is expressed by intervening PI3 K/Akt pathway,reducing expressions of HER-2,PI3 K and p-Akt in pathway.