Effects of C3G Knockout on Proliferation and Apoptosis in H9C2 Cardiomyocytes

Abstract: The NT CRISPR/Cas9 (non-target)， C3G CRISPR/Cas9 plasmids were constructed and packaged into lentiviruses respectively. H9C2 cardiomyocytes were infected with above lentiviruses respectively to investigate the effects of C3G [Crk SH3-domain-binding guanine nucleotide exchange factor] knockout on proliferation and apoptosis in H9C2 cardiomyocytes and their underlying mechanisms. The experiments were divided into NT CRISPR/Cas9， C3G CRISPR/Cas9， NT CRISPR/Cas9+Hypoxia and C3G CRISPR/Cas9+Hypoxia groups. C3G mRNA was detected by RT-PCR. C3G， p-ERK1/2， Bcl-2 and Bax proteins were tested by Western blot. Cell proliferative rate was examined by CCK-8. Apoptotic rate was determined by flow cytometry. The results showed that the expression of C3G mRNA and protein were absent in C3G CRISPR/Cas9 and C3G CRISPR/Cas9+Hypoxia groups. Compared with the NT CRISPR/Cas9 and NT CRISPR/Cas9+Hypoxia groups， the expression of p-ERK1/2 and Bcl-2 proteins and cell proliferative rate were decreased (P<0.05)， while the expression of Bax protein and the apoptotic rate were increased in the C3G CRISPR/Cas9 and C3G CRISPR/ Cas9+Hypoxia groups (P<0.05). Compared with the NT CRISPR/Cas9 group， the expression of C3G mRNA and protein (P<0.05)， p-ERK1/2 and Bcl-2 proteins and the proliferative rate were decreased in the NT CRISPR/ Cas9+Hypoxia group (P<0.05)， while the expression of Bax protein and cell apoptotic rate were increased (P<0.05). Compared with the C3G CRISPR/Cas9 group， the expression of p-ERK1/2 and Bcl-2 proteins and the proliferative rate were decreased in the C3G CRISPR/Cas9+Hypoxia (P<0.05)， while the expression of Bax protein and cell apoptotic rate were increased (P<0.05). The above results demonstrated that C3G knockout can inhibit the proliferation and promote the apoptosis of H9C2 cardiomyocytes through regulation of p-ERK1/2， Bcl-2 and Bax.

CSTR: 32200.14.cjcb.2018.06.0014