RegIV potentiates colorectal carcinoma cell migration and invasion via its CRD domain

The RegIV gene is differentially expressed in cancer and noncancerous cells. In this study, we investigated the role of RegIV and its CRD domain in the invasion and migration of human colorectal carcinoma LoVo cells. Recombinant plasmids, pcDNA3.1-RegIV and pcDNA3.1-RegIV CRD, were constructed and transfected into LoVo cells. The in vivo RegIV and RegIV CRD mRNA and protein levels were then analyzed by reverse transcriptase polymerase chain reaction and inununocytochemistry, respectively. Cell proliferation was measured by MTT (3-(4,5-dimethylthiazol-2-y1)-2,5-diphenyltetrazolium bromide) and flat plate colony-forming assays, and apoptosis was measured by flow cytometry. Cell migration and invasion were assessed with a Transwell chamber. A high level of RegIV and RegIV carbohydrate-recognition domain (CRD) expression was demonstrated in RegIV and RegIV CRD transfected cells. There were no differences in cell proliferation, colony formation, and apoptosis between the LoVo/RegIV and untreated LoVo cells (P > 0.05). However, the LoVo/RegIV cells, but not LoVo/RegIV CRD cells, displayed greater migration and invasion abilities compared to the empty vector and untreated LoVo cells. The migrated cell numbers of the LoVo/RegIV, LoVo/RegIV CRD, LoVo/empty vector, and control LoVo groups were 247.00 +/- 10.61, 146.27 +/- 6.81, 151.16 +/- 7.18, and 149.65 +/- 6.53 cells per field, respectively (P < 0.05). The invasion cell numbers of the four groups were 57.00 +/- 3.00, 31.53 +/- 2.72, 30.3 +/- 2.07, and 32.16 +/- 2.30 cells per field, respectively. RegIV enhances LoVo cell migration and invasion, and its CRD domain is critical for these effects. (C) 2010 Elsevier Inc. All rights reserved.