Effect of ART 1 gene silencing by RNA interference on the proliferation of mouse colon carcinoma cells and its possible mechanism
10.3781/j.issn.1000-7431.2012.12.001
- Author:
Jian-Xia XU
1
Author Information
1. Department of Pathology
- Publication Type:Journal Article
- Keywords:
ART1;
Cell proliferation;
Colonic neoplasms;
Glycosyltransferases;
Rho factor;
RNA interference
- From:
Tumor
2012;32(12):949-954
- CountryChina
- Language:Chinese
-
Abstract:
Objective: To investigate the effect of ART 1 (arginine-specific adenosinediphosphateribosyltransferase 1) gene silencing by shRNA (short hairpin RNA) interference on the proliferation ability of mouse colon cancer CT26 cells, and to explore its possible mechanism. Methods: It was confirmed that ART1 expression existed in CT26 cells by immunofluorescence assay. Lentivirus of ART1-shRNA was infected into mouse colon carcinoma CT26 cells. The CT26 cells uninfected or infected with a negative NC-shRNA (control-shRNA) served as the controls. The expression of ART1 mRNA was detected by RTPCR, and the expressions of ART1, RhoA, c-myc, and c-fos proteins were examined by Western blotting. The cell proliferation in each group was measured by CCK8 (cell counting kit-8) assay. Results: It was determined that ART1 expression existed in the CT26 cells. Lentivirus of ART1-shRNA or NC-shRNA was infected into CT26 cells successfully, and the CT26 cell line with stable low-expression of ART1 was successfully established. Compared with CT26 cells infected with NC-shRNA lentivirus or those were un-infected, the expression of ART1 mRNA was significantly reduced in CT26 cells infected with ART1- shRNA lentivirus (P < 0.01), and the protein expression levels of ART1, RhoA, c-myc, and c-fos were all obviously decreased (P < 0.01). The inhibition rate of cell proliferation of CT26 cells infected with ART1-shRNA lentivirus was markedly increased compared with the control groups (P < 0.01). Conclusion: RNA interference targeting ART 1 gene can inhibit the proliferation ability of mouse colon carcinoma CT26 cells. This effect probably associates with the down-regulation of the expressions of RhoA and its downstream effectors c-myc and c-fos after silencing the expression of ART 1 gene. Copyright © 2012 by TUMOR.
