Objective: To investigate the effect of siRNA-induced silencing of carcinoembryonic antigen-related cell adhesion molecule 1 (CEACAM1) on proliferation and apoptosis in human glioma SHG44 cells.
Methods: Three pairs of specific siRNA targeting CEACAM1 were designed and synthesized, and then transiently transfected into SHG44 cells via cationic liposome transfection method. Transfection efficiency was examined by flow cytometry (FCM). Real-time quantitative PCR (qRT-PCR) and Western blotting were respectively used to detect CEACAM1 expression at mRNA and protein levels, and the proliferation ability and apoptosis of SHG44 cells after transfection were assessed by CCK-8 assay and FCM in combination with annexin V-FITC/PI staining, respectively. The expression levels of cleaved caspase-3 and cleaved poly(ADP-ribose)polymerase (PARP) proteins were detected by Western blotting.
Results: The efficiency of CEACAM1 siRNA transfection into human SHG44 cells were 85%. Forty-eight hours after the three pairs of specific CEACAM1 siRNA were transfected into SHG44 cells, qRT-PCR and Western blot results showed that the expression of CEACAM1 mRNA and protein were significantly inhibited compared with those in the blank control and the negative control groups, and the most significant interference effect was CEACAM1-siRNA3. The proliferation of SHG44 cells was inhibited and the apoptosis rate was raised by the CEACAM1-siRNA transfection. The expression levels of cleaved caspase-3 and cleaved PARP proteins were up-regulated after silencing of CEACAM1.
Conclusion: The siRNA-mediated CEACAM1 silencing can inhibit the proliferation and promote the apoptosis in human glioma SHG44 cells. CEACAM1 gene may be used as a potential therapeutic target of glioma.