Effects of secreted frizzled-related proteins 2 and β-catenin on proliferation, migration and invasion of colorectal cancer cells

Abstract

Objective To observe the effects of secreted frizzled-related proteins 2 (sFRP2) and β-catenin on proliferation, migration and invasion of colorectal cancer cells. Methods The pcDNA3.1 plasmid containing sFRP2 gene sequence was constructed to overexpress sFRP2 in HCT116 by transfection. The expression level of sFRP2 in HCT116 cells after transfection was detected by Western blotting. Cell counting kit-8 (CCK-8) assay was used to detect the effect of sFRP2 upregulation on the proliferation of colorectal cancer cells; Wound-healing assay was used to detect the effect of sFRP2 up-regulation on the migration of colorectal cancer cells; Transwell assay was used to detect the effect of sFRP2 up-regulation on the invasion ability of colorectal cancer cells. Results After plasmid transfection, the expression of sFRP2 and β-catenin significantly increased. The ratio of sFRP2 protein to β-actin was 0.62±0.03 vs. 0.21±0.02 (t=25.430, P=0.001), and the ratio of β-catenin to β-actin was 0.32±0.02 vs. 0.17±0.01 (t=15.000, P=0.001), and the difference was statistically significant. The absorbance values at 12, 24, 36, 48 and 60 h in sFRP2 transfection group, control group and empty plasmid group were measured to be 0.213±0.015 vs. 0.215±0.018 vs. 0.218±0.012, 0.345±0.053 vs. 0.351±0.041 vs. 0.362±0.038, 0.382±0.058 vs. 0.413±0.051 vs. 0.433±0.048, 0.441±0.079 vs. 0.503±0.033 vs. 0.534±0.053, and 0.482±0.034 vs. 0.594±0.045 vs. 0.628±0.062. As compared with the control group and the empty plasmid group, the cell proliferation rate in sFRP2 transfection group was significantly decreased (12 h: t=0.016, P=0.988 and t=0.041, P=0.970; 24 h: t=0.155, P=0.884 and t=0.452, P=0.675; 36 h: t=0.695, P=0.525 and t=1.173, P=0.306; 48 h: t=1.254, P=0.278 and t=1.693, P=0.166; 60 h: t=3.440, P=0.026 and t=3.576, P=0.023). The difference in the distances between the both sides of scratches at 0 h and 48 h in the blank group was used as control. The distance in the blank group, empty plasmid group, and sFRP2 transfection group was 1.00±0.00, 0.91±0.06, and 0.61±0.04, respectively. The distance in sFRP2 transfection group was farther than blank group and empty plasmid group, and the migration speed was significantly slower (t=16.890, P=0.001 and t=7.206, P=0.002); The number of transmembrane cells in the sFRP2 transfection group was (75.69±7.19) cells, significantly less than that in the control group [(195.39±8.68) cells, t=25.459, P=0.001], indicating the invasive ability of the cells was significantly decreased in the sFRP2 transfection group. Conclusion The up regulation of sFRP2 significantly inhibited the proliferation, migration and invasion of colorectal cancer cell line HCT116. Key words: Colorectal cancer; Secreted Frizzled-related proteins 2; β-catenin; Proliferation; Migration; Invasion