山慈菇介导VEGF-A血管生成抑制乳腺癌细胞上皮-间质转化的实验研究

Rhizoma Pleionis inhibits of breast cancer epithelial-mesenchymal transition by VEGF-A related angiogenesis

目的探讨山慈菇在乳腺癌细胞内介导血管内皮生长因子A(VEGF-A)相关血管生成抑制乳腺癌上皮-间质转化(EMT)。方法首先通过噻唑蓝(MTT)法检测不同浓度山慈菇(0. 2、0. 4、0. 6、0. 8、1. 0 g/ml)对乳腺癌细胞MDA-MB-231和4T1增殖活力的影响;然后选取特定浓度山慈菇,通过Transwell试验检测其对乳腺癌细胞转移及侵袭能力的影响,测定人脐静脉上皮细胞(HUVEC)毛细管壁形成以检测山慈菇对细胞血管生成的影响;最后Western blot法检测山慈菇对EMT相关蛋白及血管生成相关分子的表达,进行统计学分析。结果 MTT结果显示,山慈菇在MDA-MB-231和4T1细胞中剂量依赖性抑制细胞增殖,0. 8 g/ml和1. 0 g/ml山慈菇对细胞增殖影响与对照组比较差异有统计学意义(P <0. 05);Transwell检测提示山慈菇可抑制乳腺癌细胞的侵袭,抑制HUVECs细胞血管壁的形成(P <0. 05)。Western blot法证实山慈菇处理乳腺癌细胞后,增加EMT相关蛋白E-钙黏蛋白(E-cadherin)表达水平(P <0. 05),并降低N-钙黏蛋白(N-cadherin)、波形蛋白(Vimentin)和锌指转录因子(Snail)的表达水平(P <0. 05);山慈菇处理后血管生成相关分子缺氧诱导因子1α(HIF-1α)、VEGF-A和血管内皮生长因子受体2(VEGFR-2)的水平显著降低(P <0. 05)。结论山慈菇通过影响VEGF-A相关血管生成从而抑制乳腺癌上皮-间质转化,抑制乳腺癌的侵袭转移。

Objective To investigate the Rhizoma Pleionis mediating vascular endothelial growth factor A( VEGF-A)-related angiogenesis in breast cancer cells to inhibit epithelial-mesenchymal transition( EMT) in breast cancer. Methods Firstly,the proliferation activity of MDA-MB-231 and 4 T1 of breast cancer cells was detected by methyl thiazolyl tetrazolium( MTT) method at different concentrations( 0. 2 g/ml,0. 4 g/ml,0. 6 g/ml,0. 8 g/ml,1. 0 g/ml);secondly,the specific concentration of Rhizoma Pleionis was selected and tested by Transwell test for its effect on breast cancer cell metastasis and invasion. The formation of human umbilical vein endothelial cells( HUVEC) capillary wall was measured to detect the effect of Rhizoma Pleionis on cell angiogenesis. The EMT-related protein and angiogenesis-related molecules were detected by Western blot. Results MTT results showed that Rhizoma Pleionis of 0. 8 g/ml and 1. 0 g/ml can inhibit the MDA-MB-231 and 4 T1 cells proliferation in a dose-dependent manner,with statistically significant difference( P < 0. 05). Rhizoma Pleionis can inhibit the invasion of breast cancer cells and inhibit the formation of blood vessel walls in HUVECs. Western blot confirmed that the expression level of EMT-related proteins: E-cadherin was significantly increased and the expression level of N-cadherin,Vimentin and Snail was significantly decreased after treatment with Rhizoma Pleionis( P < 0. 05);and the level of the related molecule hypoxia inducible factor-1α( HIF-1α),VEGF-A and vascular endothelial growth factor receptor-2( VEGFR-2) was significantly reduced( P < 0. 05). Conclusions Rhizoma Pleionis inhibits the epithelial-mesenchymal transition of breast cancer by influencing the angiogenesis of VEGF-A,and inhibits the invasion and metastasis of breast cancer.