The Research Of KPNA2’s Expression In Osteosarcoma Tissue And It’s Effect On Cellular Proliferation In Osteosarcoma Cell Line

Background:Karyopherin α2,belongs to the kar yopherin α family,plays a critical role in tumorigenesis and tumor progression.KPNA2 has been reported in many cancers;however,nothing is currently known about the effects of KPNA2 on osteosarcomas.Objective:This study aimed to investigate the differential KPNA2 protein and mRNA expression in human osteosarcoma tumor cells and normal bone tissue.We also want to determine if KPNA2 can influence the proliferation and apoptosis of the Saos-2osteosarcoma cell line.Methods:Immunohistochemistry(IHC)was used to investigate KPNA2 protein expression in mormal bone tissues and human osteosarcoma tissues.Real-time quantitative PCR(qPCR)was used to detect the expression of KPNA2 mRNA levels in four osteosarcoma cell lines,and lentivirus-mediated short hairpin RNAs(shRNAs)were used to knock down KPNA2 expression in Saos-2 cells.MTT assay and Multiparametric high-content screening(HCS)were used to measure cell proliferation and growth,respectively.Flow cytometry was conducted to detect cell cycle distribution and apoptosis.Results:The results revealed that KPNA2 expression levels were significantly higher in osteosarcoma tissues were remarkably higher compared with the levels in normal bone tissues;furthermore,KPNA2 m RNA was also highly expressed among fourosteosarcoma cell lines.After transducing Saos-2 cells with KPNA2-shRNA lentivirus,the proliferative rate was evidently decreased compared to the negative control(NC)lentivirus group(P<0.05).The flow cytometry results indicated that compared with control group,the S phase and G2/M phase cells were apparently increased after transducing Saos-2 cells were transduced with KPNA2-shRNA lentivirus,suggesting that KPNA2 may arrest cell cycle progression and regulate the growth of Saos-2 cells.The apoptosis results indicated that KPNA2-shRNA cells apoptotic rate was 13.38±0.48%,which was higher than the rate of cells in the control group(5.13 ±0.33%).Conclusion:This study showed that KPNA2 is higher exepression in osteosarcoma tissues and that reduced KPNA2 mRNA levels inhibit proliferation and promote apoptosis in an osteosarcoma cell line.