Effects of DNAJB 11 gene-silencing on proliferation, cell cycle and apoptosis of human hepatocellular carcinoma SMMC7721 cells / 肿瘤

ABSTRACT

Objective: To investigate the effects of DNAJ homolog subfamily B member 11 (DNAJB 11) gene-silencing on proliferation, cell cycle and apoptosis of human hepatocellular carcinoma cell line SMMC7721. Methods: The recombinant lentiviral vector pCDH-Puro/DNAJB11-shRNA carrying the specific shRNA targeting DNAJB 11 gene was established. The SMMC7721 cells were infected with high infective lentivirus pCDH-Puro/DNAJB11-shRNA. Then the proliferation of SMMC7721 cells was detected by CCK-8 method. The expression levels of DNAJB11, proliferating cell nuclear antigen (PCNA) and caspase-3 mRNAs and proteins in SMMC7721 cells were detected by real-time fluorescent quantitative PCR and Western blotting, respectively. The cell cycle distribution and the apoptosis rate of SMMC7721 cells were analyzed by FCM. Results: The pCDH-Puro/DNAJB11-shRNA was constructed successfully. The proliferation of SMMC7721 cells was significantly inhibited after infection with pCDH-Puro/DNAJB11- shRNA (P<0.05). In SMMC7721 cells infected with pCDH-Puro/DNAJB11-shRNA, the expressions of DNAJB11 mRNA and protein were silenced effectively (both P<0.05). After DNAJB 11 gene-silencing, the expressions of caspase-3 mRNA and protein in SMMC7721 cells were up-regulated (both P<0.05), while the expressions of PCNA mRNA and protein were down-regulated (both P<0.05). Furthermore, the cell cycle was arrested in G1 phase (P<0.01), and the apoptosis rate was significantly increased (P<0.01). Conclusion: The DNAJB 11 gene-silencing can effectively suppress the proliferation of SMMC7721 cells, and promote their apoptosis. These effects may be related to downregulation of PCNA expression and upregulation of caspase-3 expression in SMMC7721 cells.