Experimental Study Of OPN Gene Silencing Mediated By SiRNA In Laryngeal Squamous Cell Carcinoma

objective :We try to look for a new approach of gene therapy of LSCC by synthe -sizing and filtrating effective OPN-siRNA in vitro, and checking the inhibitory effect of gene silencing mediated by OPN-siRNA for LSCC in vivo. Methods: emi-quantita -tive analysis of the level of OPN-mRNA expression was performed using the RT PCR technique. The level of OPN protein expression was analyzed with Western - Bl -oting technique, the proliferation index (PI) was calculated. The cell growth and vig -our was detected by MTT Assay .The models of subcutaneous LSCC tumor in nude mi -ce were established with Hep-2 cells. The OPN-siRNA lentivirus have been discontin -ueously injected directly into the established tumor, The changes of weight and size of the tumors and the mice body weight during the therapy were calculated respective-ly. Results: The content of protein of OPN-1 in the OPN-siRNA lentivirus transfect -ed Hep-2 cells was obviously reduced compared with blank control cells.The content of mRNA of OPN in the OPN-siRNA lentivirus transfected Hep-2 cells was obvious -ly reduced compared with blank control cells.PI staining in the OPN-siRNA lentivir -us transfected cell showed 30%-35% of apoptotic cells in Hep-2 cells.MTT Assay res -ult display, Hep-2 cells' vigour decrease obviously after be transfected by OPN-siR -NA lentivirus. The growth of Tumors in the OPN-siRNA lentivirus injected nude mi -ce being inhibitted obviously through tumor growth curve. The weight and volume of transplant tumors in the OPN-siRNA lentivirus injected nude mice were significantly reduced compare with negative siRNA control nude mice and blank control nude mic -e. The weight of transplant tumors was reduced 65.37% compared with blank contro -l.Conclusion:The OPN protein remarkably up regulated in the Hep-2 cells may invol -ve in LSCC carcinogenesis.Eligible 21nt siRNAs synthesized in vivo with lentiviral vector mediated can be highly effectively transfected into LSCC cell, and induced po -st-transcriptional gene silencing.OPN gene silencing mediated by siRNA significant ly suppresses the growth of Hep-2 cell arrest in vitro,30%-35% of apoptotic cells in Hep-2 cells ;and significantly achieve anti-tumor effect in subcutaneous transplant LSCC tumor in nude mice in vivo.