Signal transduction mechanism of phospholipase C?1 in colorectal cancer cells / 解放军医学杂志

ABSTRACT

Objective To investigate signal transduction mechanism of phospholipase C?1 (PLC?1) in colorectal cancer cells. Methods Gel electrophoresis mobility shift assay (EMSA), immunocytochemistry, zymography and RT-PCR were performed to investigate the function of PLC?1 on nuclear factor-Kappa B (NF-?B), matrix metalloproteinase-2 (MMP-2) and tissue inhibitor of metalloproteinase-2 (TIMP-2) in LoVo cell. Results Compared with control group, nuclear positive rate of cells treated with epidermal growth factor (EGF) increased significantly (from 26.91%?2.84% to 40.83%?4.36%), while that of cells treated with 2.5mol/L U73122 decreased to 12.20%?1.89%. Meanwhile, pretreatment with 2.5mol/L U73122 before EGF treatment decreased nuclear positive rate of cells from 40.83%?4.36% to 18.21%?1.34%. The results of EMSA further verified that PLC?1 can regulate the activity of NF-?B. RT-PCR results showed that EGF, PLC?1 or NF-?B had no significant effect on the expression of MMP-2, TIMP-2 at mRNA level. Furthermore, zymography indicated that the activity of MMP-2 did not change dramatically by EGF, PLC?1 or NF-?B. Conclusion These data suggested that EGF-PLC?1-NF-?B signaling pathway was operative in LoVo cell, but MMP-2 and TIMP-2 may not be regulated by EGFR-PLC?1-NF-?B signaling pathway.