| Objective:To investigate the inhibitory effect on proliferation by grape procyanidins (GPC) and the regulation of estrogen receptor(ER) in human breast cancer cells.Methods:1.MCF-7 cells were cultured with GPC of different doses.The experiment was divided into tumor control group(GPC 0μg/mL) and five doses of GPC groups(GPC 10, 50,100,150 and 200μg/mL).The proliferation activity of MCF-7 cells was determined by MTT assay at different time(6h,12h,24h,48h and 72h).2.The experiment was divided into tumor control group(GPC 0μg/mL) and three doses of GPC groups(GPC 50,100 and 200μg/mL).After cultured with GPC of different doses for 24h,the expression levels of the proliferating cell nuclear antigen(PCNA),Bcl-2, Bax and ER protein in MCF-7 cells were measured by immunohistochemistry method.3.MCF-7 cells were cultured with GPC and estrogen antagonist ICI182.780(ICI).The experiment was divided into four groups:tumor control group(GPC 0μg/mL and ICI 0 mol/L),ICI control group(ICI 10-6mol/L),dose of GPC group(GPC 200μg/mL) and GPC +ICI group(GPC 200μg/mL and ICI 10-6mol/L).After cultured for 24h,the proliferation activity was determined by MTT assay;the expression levels of PCNA,Bcl-2 and Bax protein were measured by immunohistochemistry method.Results:1.At the same time,with the increasing dose of GPC,the proliferation activity levels of 10,50,100,150 and 200μg/mL GPC groups decreased gradually(P＜0.05).At the same dose of GPC,with the increasing time,the proliferation activity levels of 100,150 and 200μg/mLGPC groups decreased gradually(P＜0.05).All above the results showed that GPC could inhibit proliferation activity of human breast cancer cells in a significant dose-and time-dependent manner.2.①Compared with tumor control group,the expressions of PCNA in 50,100 and 200μg/mL GPC groups were significantly decreased(P＜0.05).There was significant difference between 50,100 and 200μg/mL GPC groups(P＜0.05).②The expression of Bcl-2 protein in 200μg/mL GPC group was significant lower than that of tumor control group(P＜0.05),but the expression of Bax protein in 200μg/mL GPC group was significant higher than that of tumor control group(P＜0.05).③The expressions of ER protein in 100 and 200μg/mL GPC groups were (86.22±5.86)%and(79.34±4.79)%respectively.Compared with tumor control group (93.90±6.07)%,all the results mentioned were significantly decreased(P＜0.05).The expression of ER protein in 200μg/mL GPC group was significant lower than that of 100μg/mL GPC group(P＜0.05).3.①The levels of proliferation activity in ICI control group and dose of GPC group were 0.346±0.015 and 0.234±0.011 respectively.Compared with tumor control group (0.373±0.009),all the results mentioned were significantly decreased(P＜0.05).The level of proliferation activity of GPC+ICI group was 0.293±0.008,which significantly increased by 0.059 compared with dose of GPC group and significantly decreased by 0.053 compared with ICI control group(P＜0.05).Compared with tumor control group,the expressions of PCNA in ICI control group and dose of GPC group were significantly decreased(P＜0.05).The expression of PCNA in GPC+ICI group was significant higher than that of dose of GPC group and significant lower than that of ICI control group(P＜0.05).③Compared with dose of GPC group,the expression of Bcl-2 protein in GPC+ICI group was significant increased and the expression of Bax protein was significant decreased(P＜0.05).Conclusion:1 GPC can inhibit the proliferation and promote the apoptosis of human breast cancer cells effectively.2 The regulation of ER is closely related to the inhibitory effect of GPC on human breast cancer cells.3 The probable mechanism is that GPC can reduce the expression of ER protein and block the combination of estrogen and ER to inhibit the growth of breast cancer cells.4 There are other pathways of the inhibitory effect of GPC on human breast cancer cells except the ER pathway. |
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