Study On Anti-Breast Cancer Activity And Mechanism Of A Novel Ruthenium Complex

Breast Cancer(BC),a clinically common tumor,occurs in the epithelial tissue of human Breast,which seriously threatens human health and even life safety.Ruthenium complexes have good anticancer activities due to their unique structural characteristics,such as high selectivity,easy to be absorbed by cancer cells,less toxicity than platinum and significant anticancer effect.In this study,the constructed breast cancer MCF-7 cell line with high expression of Sigma-1 receptor(MCF-7-sg1 for short)and breast cancer MCF-7 and MDA-MB-231 cells were used to explore the anti-breast cancer activity and the mechanism of the new Ruthenium complex.The main results are listed as follows:(1)MCF-7 cells with high expression of sigma-1 receptor were constructed.MCF-7 was transfected with pc DNA3.1-Sigma-1 vector plasmid,and genomic DNA was extracted from the surviving cells after screening.After PCR amplification,agarose electrophoresis was performed,and proteins were prepared for western-blot.Finally,MCF-7 cell line of breast cancer with high expression of Sigma-1 receptor was successfully constructed,named MCF-7-sg1.(2)Ruthenium complex can inhibit the proliferation of breast cancer cells.MCF-7,MCF-7-sg1 and MDA-MB-231 cells were treated with Ruthenium complex at different concentrations(0.0,1.0,5.0 and 10.0 ?mol/L).The results showed that Ruthenium complex inhibited the proliferation of three breast cancer cells in a dose-dependent manner.Ruthenium complex significantly inhibited MDA-MB-231 cells more than MCF-7 line,and significantly inhibited MCF-7-sg1 more than MCF-7cells(groups treated with 5.0 and 10.0 ?mol/L).(3)Ruthenium complex can inhibit the migration ability of breast cancer cells.MCF-7,MCF-7-sg1 and MDA-MB-231 cells were treated with Ruthenium complex at different concentrations(0.0,1.0,5.0,and 10.0 ?mol/L)for 24,48,and 72 h.The results showed that Ruthenium complex could inhibit the migration of three types of breast cancer cells in a concentration dependent manner.Compared with the control group,Ruthenium complex at 3 concentrations(1.0,5.0,10.0 ?mol/L)significantly inhibited the migration of MCF-7-sg1 and MDA-MB-231 cells at 3 time points(24,48,and 72 h),while MCF-7 cells were significantly inhibited only at 10.0 ?mol/L.The inhibition of MCF-7-sg1 and MDA-MB-231 migration was found to be significantly stronger than that of MCF-7 cells only at 72 h after treatment with 1?mol/L Ruthenium complex.(4)Ruthenium complex can inhibit the invasion ability of MCF-7 and MCF-7-sg1 cells of breast cancer.Breast cancer cell lines MCF-7,MCF-7-sg1 and MDA-MB-231 were treated with Ruthenium complex at different concentrations(1.0,5.0,10.0 ?mol/L)for 24 h,and Transwell invasion assay was used to detect the invasion ability of these three cell lines.The results showed that Ruthenium complex could inhibit the invasion ability of MCF-7 and MCF-7-sg1 cells in a concentration-dependent manner,but had little effect on the invasion ability of MDA-MB-231 cells.Ruthenium complex significantly inhibited the invasion of MCF-7-sg1 cells more than MCF-7 cells(treated with 5.0 and 10.0 ?mol/L).(5)Ruthenium complex had no significant effect on the expression of Matrix Metalloproteinases(pro-MMP-9 and MMP-2)in MCF-7 and MCF-7-sg1 cells of breast cancer.The expression of matrix metalloproteinases in MCF-7,MCF-7-sg1 and MDA-MB-231 cells was detected by gelatinase assay after 24 h treatment with Ruthenium complex at different concentrations(1.0,5.0,10.0 ?mol/L).Two Matrix Metalloproteinases were detected,namely pro-MMP-9(92 k Da)and MMP-2(66 k Da).The results showed that the expression of Matrix Metalloproteinases(pro-MMP-9 and MMP-2)in MCF-7 and MCF-7-sg1 cells did not change significantly with the increase of Ruthenium complex treatment concentration,while only weak expression of MMP-2 was detected in MDA-MB-231 cells.In conclusion,the Ruthenium complex can inhibit the activity,migration and invasion ability of breast cancer cells MCF-7,MCF-7-sg1 and MDA-MB-231(among which,it has little effect on the invasion ability of MDA-MB-231 cells).The Ruthenium complex inhibit the migration and invasion of breast cancer cells not by affecting the expression of Matrix Metalloproteinases(pro-MMP-9 and MMP-2).In addition,MCF-7 cells transfected with Sigma-1 receptor can improve the effect of the Ruthenium complex.