慢病毒介导shRNA抑制IL1R2基因对人骨肉瘤细胞U-2 OS增殖功能的影响

IObjectiv: This study aimed to construct a lentivirus harboring shRNA against IL1R2 and investigate its effect on the proliferation of human osteosarcoma cell line U-2 OS. Methods: Two pairs of shRNA oligos (S1 and S2) and a non-coding sequence (shCon) based on IL1R2 gene were constructed and identified, then were transferred into 293T cell to measure the titer. After the best MOI and the better target were tested, the proliferations of U-2 OS cells before and after the infection of lentiviral vectors were compared by the growth curve, plate clony formation assay and flow cytometry. Results: The lentiviruses were identified as designed. The titer of lentivirus was 1×107 TU/ml, and the best MOI was 20. The S1 had a higher efficiency than S2.Significant differences were discovered that both in growth curve measurement and plate clony formation assay, cells infected with S1 had lower proliferation than the cells infected with shCon. In flow cytometry, cells infected with S1 were significantly less in phase G0/ G1 and more in phase S and G2 /M compared to the cells infected with shCon. Conclusion: The proliferation and clonality of human osteosarcoma cell line U-2 OS are significantly down-regulated as infected with lentiviruses harboring shRNA against IL1R2. Il1R2 may have oncogenic potential, therefore it is possible to become a new target for genetic therapy against osteosarcoma.