Effects of nephroblastoma overexpressed gene on proliferation, apoptosis and migration of human osteosarcoma 143B cells / 肿瘤

ABSTRACT

Objective: To investigate the effects of nephroblastoma overexpressed (NOV) gene on proliferation, apoptosis and migration of human osteosarcoma 143B cells in vitro, and to explore its underlying mechanism. Methods: The osteosarcoma cell line with low expression of NOV was screened out by semiquantitative reverse transcription-polymerase chain reaction (RT-PCR) and Western blotting. The 143B cells were infected with recombinant adenoviruses AdNOV and AdGFP (as a negative control) and cultured for 24 h. Then the infection efficiencies of recombinant adenoviruses AdNOV and AdGFP in 143B cells were observed under a fluorescence microscope. The expression levels of NOV mRNA and protein were detected by semi-quantitative RT-PCR and Western blotting, respectively. The effects of NOV overexpression on proliferation, apoptosis and migration of 143B cells were measured by MTT, flow cytometry (FCM) and Transwell migration test, respectively. The expression levels of NOV, B cell lymphoma-2 (Bcl-2) and Bcl-2-associated X protein (Bax) in 143B cells were examined by semi-quantitative RT-PCR and Western blotting. The expression levels of phospho-p38 (p-p38), total p38, phospho-c-Jun N-terminal kinase (p-JNK) and total JNK were detected by Western blotting. Results: The expression level of NOV was lowest in 143B cells (P 0/G1 phase (P < 0.05). As compared with the control group, NOV overexpression increased the expession level of Bax (P < 0.001) and down-regulated the expression level of Bcl-2 (P < 0.05) in 143B cells. At the same time, the expression levels of p-p38 and p-JNK were elevated significantly in 143B cells overexpressing NOV (P < 0.001). Conclusion: NOV overexpression induced by adenovirus can inhibit the proliferation of 143B cells and promote the apoptosis and migration of 143B cells in vitro. The p38/mitogenactivated protein kinase (MAPK) and JNK/MAPK signaling pathways may be involved in this process.