Effects of recombinant adenovirus Ad-PERK siRNA on the proliferation and apoptosis of human chondrosarcoma SW1353 cells in endoplasmic reticulum stress / 肿瘤

ABSTRACT

Objective: To investigate the effects of small interfering RNA (siRNA) targeting protein kinase-like endoplasmic reticulum kinase (PERK) gene recombinant adenovirus on the proliferation and apoptosis of human chondrosarcoma SW1353 cells in endoplasmic reticulum stress (ERS). Methods: Recombinant adenovirus Ad-PERK siRNA targeting PERK gene was constructed, and then it was infected into SW1353 cells. The expression levels of PERK mRNA and protein in SW1353 cells after infection with Ad-PERK siRNA were detected by reverse transcription-PCR (RT-PCR) and Western blotting, respectively. ERS model was induced by tunicamycin (TM). Under the ERS condition, the proliferation of SW1353 cells after infection with Ad-PERK siRNA was determined by MTT assay, the cell cycle distribution and apoptosis were measured by flow cytometry (FCM), the change of the microscopic structure was observed under a transmission electron microscope, and the expression levels of cleaved caspase 3, caspase 12, CCAAT/enhancer-binding protein homologous protein (CHOP) and phospho-c-Jun-N-terminal kinase (p-JNK) proteins were detected by Western blotting. Results: The recombinant adenovirus Ad-PERK siRNA was successfully constructed. The expression levels of PERK mRNA and protein in SW1353 cells after infection with Ad-PERK siRNA were decreased (P < 0.05). Under the ERS condition, the proliferation of SW1353 cells after infection with Ad-PERK siRNA was promoted (P < 0.05). The ratio of S phase in Ad-PERK siRNA infection group was higher than those in negative control adenovirus Ad-RFP infection group (as a negative control) and non-infection group (as a blank control) (both P < 0.05). The apoptotic rate of SW1353 cells after infection with Ad-PERK siRNA was lower than those of the negative control and the blank control groups (P < 0.05). The apoptosis structures of SW1353 cells in the negative control and the blank control groups were clearly observed under a transmission electron microscope. The expression levels of cleaved caspase 3, caspase 12, CHOP and p-JNK proteins in SW1353 cells of Ad-PERK siRNA infection group were lower than those of the negative control and the blank control groups (P < 0.05). Conclusion: The recombinant adenovirus Ad-PERK siRNA can promote the proliferation of SW1353 cells and inhibit the apoptosis under the ERS condition.