目的探讨穿心莲内酯(Andro)对人乳腺癌细胞MCF-7的增殖及细胞周期的影响,为其应用于乳腺癌的治疗提供理论基础。方法将实验分为对照组及Andro组,Andro组加入终浓度为10、20、40、60、80、100μmol/L的Andro[Andro粉末先用DMSO溶解,再用培养基稀释至细胞处理浓度,控制DMSO的含量为0.1%(v/v)],对照组仅用0.1%DMSO溶剂。采用CCK8检测Andro不同浓度及不同处理时间(24、48、72 h)对MCF-7细胞增殖的影响;应用流式细胞术检测两组细胞周期分布情况;应用预胶化淀粉细胞块制作方法,将两组细胞制作成石蜡切片,应用免疫组织化学方法检测切片中MCF-7细胞增殖相关的核抗原Ki67及细胞周期相关抗原周期素D1(Cyclin D1)及周期素依赖性激酶4(CDK4)的表达。结果 CCK8检测结果显示Andro浓度为10、20、40、60、80、100μmol/L显著抑制MCF-7细胞的增殖,IC_(50)值为25μmol/L,并且在IC_(50)浓度下24、48、72 h Andro均显著抑制MCF-7细胞的增殖,并随着作用时间的延长,抑制作用越明显。细胞周期检测结果 Andro处理后G_0/G_1期细胞数量达(75.51±1.56)%,与对照组(49.16±1.35)%比较,显著升高(P<0.05),而S期的细胞数量仅为(12.69±0.87)%,与对照组(44.07±0.98)%比较,显著降低(P<0.05)。细胞块免疫组织化学结果显示对照组与Andro组Ki67的表达分别为(91.77±2.75)%、(78.05±2.61)%,显著降低(P<0.05),Cyclin D1的IOD值分别为(2.14±0.17)×10~7、(1.27±0.22)×10~7,显著降低(P<0.05),CDK4的IOD值分别为(1.14±0.014)×10~7、(1.16±0.024)×10~7,无明显差异。结论 Andro可通过抑制MCF-7细胞的增殖及抑制Cyclin D1的表达从而阻断MCF-7细胞的细胞周期进程从而发挥抗肿瘤作用,Andro不能通过影响CDK4的表达发挥阻断MCF-7细胞周期。 Objective To investigate the effects of Andrographolide on the proliferation and cell cycle of human breast cancer cell line MCF-7 and to provide a theoretical basis for its application in the treatment of breast cancer. Methods The experiment was divided into control group and Andro group, Andro group was added with 10, 20, 40, 60, 80 and 100μmol / L of Andro [Andro powder was first dissolved in DMSO and then diluted to medium concentration, The content of DMSO was controlled to be 0.1% (v / v)], while the control group only contained 0.1% DMSO. CCK8 was used to detect the effect of different concentrations of Andro on the proliferation of MCF-7 cells at different concentrations (24, 48 and 72 h). Flow cytometry was used to detect the distribution of cell cycle in both groups. The pregelatinized starch cell mass The two groups of cells were made into paraffin sections, and the expression of Ki67, cyclin D1 and cyclin-dependent kinase 4 (CDK4), a nuclear antigen associated with proliferation of MCF-7 cells, were detected by immunohistochemistry expression. Results The results of CCK8 showed that the concentration of Andro was 10, 20, 40, 60, 80 and 100 μmol / L significantly inhibited the proliferation of MCF-7 cells. The IC 50 value was 25 μmol / L and the IC 50 values ​​were 24, At 48 and 72 h, Andro significantly inhibited the proliferation of MCF-7 cells, and the more obvious the inhibitory effect was with the prolongation of action time. Cell cycle test results The cell number in G0 / G1 phase after treatment with Andro reached 75.51 ± 1.56%, which was significantly higher than that in control group (49.16 ± 1.35)% (P <0.05) 12.69 ± 0.87)%, which was significantly lower than that of the control group (44.07 ± 0.98)% (P <0.05). The results of immunohistochemistry showed that the expression of Ki67 in control group and Andro group were (91.77 ± 2.75)% and (78.05 ± 2.61)%, respectively (P <0.05), and the IOD values ​​of Cyclin D1 were (2.14 ± 0.17 (1.14 ± 0.014) × 10 ~ (7), (1.16 ± 0.024) × 10 ~ 7, respectively, which were significantly lower than those in control group (P <0.05) No significant difference. Conclusions Andro can inhibit the proliferation of MCF-7 cells and the expression of Cyclin D1, thereby blocking the cell cycle progression of MCF-7 cells to exert antitumor effects. Andro can not block the MCF-7 cell cycle by affecting the expression of CDK4.