乳岩宁方联合依西美坦对MCF-7乳腺癌细胞周期及ERα、p-Akt表达的影响

Effects of Ruyanning Prescription and Exemestane on Cell Cycle and the Expressions of ERα and p-Akt in MCF-7 Breast Cancer Cells

目的:观察中药复方乳岩宁联合依西美坦对MCF-7(michigan cancer foundation-7,MCF-7)乳腺癌细胞周期及ERα(estrogen receptorα,ERα)、p-Akt(phosphorylated-protein kinase B,p-Akt)蛋白表达的影响,探讨中药复方乳岩宁联合依西美坦抑制MCF-7乳腺癌细胞增殖的信号转导通路。方法:将40只造模成功的绝经荷瘤裸鼠随机分为模型组、依西美坦组、乳岩宁组、结合组(乳岩宁+依西美坦)。连续给药21天后脱颈处死裸鼠,完整剥离出瘤体,切开标本取材。采用碘化丙啶(propidium iodide,PI)单染法测乳腺癌细胞周期,蛋白免疫印迹法(Western blot)检测乳腺癌组织中p-Akt、ERα蛋白表达。结果:各用药组G_(0)/G_(1)(zero/first gap)期细胞比例增多,S期比例减少,与模型组比较差异有统计学意义(P<0.01);结合组G_(0)/G_(1)期细胞比例最多,S(synthesis phase)期细胞比例最少,与依西美坦组、乳岩宁组比较差异有统计学意义(P<0.05);乳岩宁组与依西美坦组比较G_(0)/G_(1)期、S期的细胞比例差异均无统计学意义(P>0.05)。模型组裸鼠瘤组织中ERα、p-Akt蛋白高表达,与各用药组相比差异有统计学意义(P<0.01);各用药组均可下调ERα、p-Akt蛋白表达,其中结合组作用更显著,与乳岩宁组、依西美坦组比较差异有统计学意义(P<0.01)。结论:乳岩宁联合依西美坦可能通过抑制组织细胞周期进程,下调癌组织中p-Akt、ERα蛋白表达来抑制PI3K-Akt信号通路的活性,实现抑制乳腺癌细胞增殖,同时中药乳岩宁对依西美坦有协同作用。

Objective:To observe the effects of herbal compound Ruyanning and exemestane on cell cycle distribution and the expressions of ERα and p-Akt protein in MCF-7 cells and investigate the signal transduction pathway of both medicine inhibiting the proliferation of MCF-7 breast cancer cells.Methods:Forty postmenopausal tumor-bearing nude mice after successful modeling were randomized into model group,exemestane group,Ruyanning group and the combination group (Ruyanning+exemestane).Nude mice were sacrificed by necking after 21 days of consecutive medication,the tumor was stripped completely,and the samples were cut to draw the materials.PI single dyeing method was used to detect cell cycle of breast cancer cells,Western blot method was adopted to measure the expressions of ERα and p-Akt protein in breast cancer tissue.Results:The proportion of cells in G_(0)/G_(1) phase increased and the proportion of cells in S phase decreased in different medication groups,when they were compared with model group,the difference had statistical meaning (P<0.01);in the combination group,the proportion of cells in G_(0)/G_(1) phase was the most,while the proportion of cells in S phase was the least,the difference was statistically significant when compared with exemestane group and Ruyanning group (P<0.05);the difference had no statistical meaning when Ruyanning group was compared with exemestane group in the proportion of cells in G_(0)/G_(1) phase and S phase (P>0.05).ERα and p-Akt protein were highly expressed in model group,and the difference was statistically significant when model group was compared with different medication groups (P<0.01);different treatment groups could down regulate the expressions of ERαand p-Akt protein,among them,the effects of the combination group were more notable,the difference had statistical meaning when the combination group was compared with Ruyanning group and exemestane group (P<0.01).Conclusion:Ruyanning and exemestane could inhibit the proliferation of breast cancer cells possibly through restraining the progression of tissue cell cycles,downregulating the expressions of ERα and p-Akt protein,therefore to restrain the activity of PI3K-Akt signaling pathway.Meanwhile,herbs Ruyanning has synergistic effect on exemestane.