Effect on cell proliferation and proteolytic enzyme of human gastric carcinoma by MUC2 antisense oligodeoxynucleotide in vitro / 第三军医大学学报

ABSTRACT

Objective To investigate the inhibition effect in vitro of mucin gene MUC2 antisense oligodeoxynucleotide (ASODN) on its gene expression and proteolytic enzyme on gastric cancer cell line. Methods Phosphorothioated MUC2 ASODN were synthesized and transfected to SGC7901 cells mediated by lipofectin. MUC2 mRNA and MUC2 protein expressed on SGC7901 cells was detected by RT-PCR and immunohistochemical method. The inhibitory effects of ASODN on proteolytic enzyme, such as cathepsin D, MMP-2 and MMP-9 protein were detected by immunohistochemical staining. The cell cycle and apoptosis were determined by flow cytometry (FCM). Results The inhibition effects peaked at 48th hour after transfection, and the inhibition rate reached 55% when the concentration of ASODN was 0.5 ?mol/L. The number of the cells treated with MUC2 ASODN was decreased as compared to the control cells. MUC2 ASODN transfection could inhibit the transition period of S phase to G2/M phase, and G2/M did not change much. The apoptosis rate was about 4.38%. As compared with blank control group, MUC2 ASODN could significantly inhibit MUC2 mRNA expression of SGC7901 cells. ASODN could downregulate the expression levels of MUC2 protein, MMP-2 and cathepsin D protein. Conclusion MUC2 ASODN transfection could specifically inhibit SGC7901 cells by downregulating the expression levels of proteolytic enzyme.