Influence of CDCA5-siRNA on proliferation and apoptosis of human hepatocarcinoma cell line HepG2
Shuzhe Chen
Abstract
Shuzhe Chen
Abstract
Objective To investigate influence of CDCA5 small interfering RNA(CDCA5-siRNA) on proliferation and apoptosis of human hepatocarcinoma cell line HepG2.Methods The HepG2 cells were divided into siRNA groups(A,B and C groups),universal scrambled negative control siRNA group(NC group) and Blank group.Three CDCA5-siRNAs with different sequences were transfected into HepG2 cells respectively.Western blotting method was used to measure expressions of CDCA5 protein levels at 72h after transfection.The cell proliferation was assessed by CCK-8 at 24,48,72,96 and 120h after transfection.The Annexin V-FITC/PI double-labeled flow cytometry was employed to measure the apoptosis at 48h after transfection.Results The expressions of CDCA5 protein significantly decreased in the CDCA5-siRNA groups(A,B and C groups) compared with NC group and Blank group(P0.05).Group B had the lowest expression rate and highest inhibition rate(89.3%).The relative proliferation rate of group B was significantly lower than those of NC group and Blank group since 48 hours after transfection(P0.05),and the lowest proliferation rate was(66.58±2.58) % at 72h in group B.The early and late apoptosis rates were(17.43±2.31) % and(22.37±2.21) % in B group,higher than those of other two groups(P0.05).Conclusion siRNA down-regulating the expression of CDCA5 gene in human hepatocarcinoma cell line HepG2 can inhibit cell proliferation and increase cell apoptosis.
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Objective To investigate influence of CDCA5 small interfering RNA(CDCA5-siRNA) on proliferation and apoptosis of human hepatocarcinoma cell line HepG2.Methods The HepG2 cells were divided into siRNA groups(A,B and C groups),universal scrambled negative control siRNA group(NC group) and Blank group.Three CDCA5-siRNAs with different sequences were transfected into HepG2 cells respectively.Western blotting method was used to measure expressions of CDCA5 protein levels at 72h after transfection.The cell proliferation was assessed by CCK-8 at 24,48,72,96 and 120h after transfection.The Annexin V-FITC/PI double-labeled flow cytometry was employed to measure the apoptosis at 48h after transfection.Results The expressions of CDCA5 protein significantly decreased in the CDCA5-siRNA groups(A,B and C groups) compared with NC group and Blank group(P0.05).Group B had the lowest expression rate and highest inhibition rate(89.3%).The relative proliferation rate of group B was significantly lower than those of NC group and Blank group since 48 hours after transfection(P0.05),and the lowest proliferation rate was(66.58±2.58) % at 72h in group B.The early and late apoptosis rates were(17.43±2.31) % and(22.37±2.21) % in B group,higher than those of other two groups(P0.05).Conclusion siRNA down-regulating the expression of CDCA5 gene in human hepatocarcinoma cell line HepG2 can inhibit cell proliferation and increase cell apoptosis.
Key concepts: Transfection, Apoptosis, Small interfering RNA, Cell growth, Annexin, Molecular biology, Flow cytometry, Cell culture