The Inhibitory Effect Of Carnosine On The Proliferation?migration And Invasion Of MCF-7 Human Breast Cancer Cells

Objectives The purpose of this study is to investigate the effect and mechanism of carnosine on the cell viability,proliferation? migration and invasion of the cultured human breast cancer MCF-7 cells.Methods The MCF-7 cells viability cultured on wells were detected by MTT reduction assay,the proliferation ability detected by plate colony formation assay,the migration and invasion ability detected by transwell migration/invasion assay,the genes expression detected by real-time PCR Assay and western blot analysis.At last,the results were statistically analyzed on the Windows platform using Graph Pad PRISM 6.0 software and SPSS 23.0 software.Results 1 The result of MTT assay showed that the viability of breast cancer cells decreased with increasing concentration after treatment with different concentrations of carnosine(12.5m M,25 m M,50 m M and 100 m M)for 24,48 and 72 hours(F=4.852,P=0.004 <0.05),and showed a concentration-dependent,but not significant in a timedependent(F=1.825,P=0.179>0.05),and there was no interaction effect between time and concentration of carnosine by statistical analysis(F=1.829,P=0.110>0.05).2 The result of plate colony formation assay showed that the number of colony formation of MCF-7 cells treated with different concentration carnosine was significantly lower than that in control group(F= 746.5,P<0.001),The number of colony formation in the 25 m M group was 135.00 ± 6.40,in the 50 m M group was 62.00 ± 7.97,in the 100 m M group was 7.4 ± 1.34 and in the control group was 223.60 ± 11.35.3 The results of Transwell migration assay showed that the average number of cells passing through the membrane in the control group and the experimental group(25,50 and 100 m M)was 313.00 ± 23.35,220.60 ± 21.54,132.40 ± 7.77 and 90.80 ± 5.50,respectively.Compared with the control group,the average cell number was decreased significantly(F=177.2,P<0.05).4 The results of Transwell invasion assay showed that the average number of cells in the blank control group and experimental group(25,50 and 100 m M)passed through the cell membrane were 258.60 ± 21.93,178.40 ± 11.26,123.40 ± 13.72 and 63.60 ± 13.50,respectively.According to statistical analysis,compared with the control group,the number of transmembrane cells in the experimental group was significantly decreased(F= 140.6,P<0.05),and the number of transmembrane cells were increased with the increase of carnosine concentration(P<0.05).5 Cell wound healing assay results showed that the healing rates in blank control group,25 m M group,50 m M group and 100 m M group were 0.46 ± 0.02,0.36 ± 0.02,0.27 ± 0.05 and 0.03 ± 0.02,respectively.And the statistical analysis showed that carnosine can significantly inhibited the wound healing rate of breast cancer cells(F= 99.20,P<0.05).6 Real-time quantitative PCR results showed that the transcriptional level of PTBP1,PKM2 and MTA1 m RNA in breast cancer cells decreased significantly(P all<0.05),however,the relative transcription level of PKM1 was significantly higher than that of the control group(P <0.05).7 Western blot results showed that the protein expression of PKM1 in the experimental group increased significantly and the expression of PTBP1,PKM2,and MTA1 decreased significantly in the experimental group(P all <0.05).Conclusions 1 Carnosine inhibited the proliferation of MCF-7 cells.The inhibitory effect of carnosine on the proliferation of MCF-7 cells might be related to down-regulation of PTBP1,PKM2 protein and up-regulation of PKM1 protein.2 Carnosine inhibited themigration and invasion of MCF-7 cells,which might be related to the down-regulation of MTA1 protein.