Functional Study On CKAP2L Promoting The Progression Of Osteosarcoma And Cisplatin Resistance

Objective: To explore the expression level of CKAP2 L in osteosarcoma and its influence on proliferation,migration,invasion and apoptosis,and further explore the expression level of CKAP2 L in cisplatin-resistant osteosarcoma cell lines and its influence on cell function.Methods:1.GEPIA and UALCAN databases were used to analyze the expression of CKAP2 L in pancancer;Use GEPIA and Kaplan-meier plotter database to explore the prognosis and survival analysis of CKAP2 L in sarcoma;The proteins related to CKAP2 L were selected by Linked Omics database and enriched and analyzed.Finally,GO and KEGG were visualized by R.2.The expression level of CKAP2 L in osteosarcoma and adjacent tissues was detected by immunohistochemistry;The expression of CKAP2 L in osteosarcoma cells and mesenchymal stem cells(MSCs)was detected by Western blot.3.Small interfering RNA(si RNA)knocks out CKAP2 L in U-2OS,and Western blot test is used to detect the knockout.4.The proliferation,migration,invasion and apoptosis abilities of si-U-2OS and U-2OS were compared through CCK-8 cell proliferation test,scratch healing test,Transwell cell invasion test,and flow cytometry detection of apoptosis.5.A drug-resistant osteosarcoma cell line U-2OS/DDP was constructed by the combination of high-dose impact of cisplatin(DDP)and low-dose maintenance.The expression of CKAP2 L in U-2OS group and U-2OS/DDP group was compared by Western blot.6.Small interfering RNA(si RNA)knocks out the CKAP2 L of U-2OS/DDP,and Western blot test is used to detect the knockout.7.The proliferation,migration,invasion and apoptosis abilities of si-U-2OS/DDP group and U-2OS/DDP group were compared through CCK-8 cell proliferation test,scratch healing test,Transwell cell invasion test,and flow cytometry detection of apoptosis.Results:1.Through GEPIA,Kaplan-meier plotter and UALCAN database,it was found that CKAP2 L was highly expressed in various tumors,and patients with high expression of CKAP2 L had poor long-term prognosis;Through the Linked Omics database and the GO and KEGG analysis of R,it was found that CKAP2 L was mainly enriched in cell cycle-related proteins.2.Western blot and immunohistochemistry results showed that CKAP2 L was low expressed in mesenchymal stem cells(MSCs)and adjacent tissues,but high expressed in osteosarcoma cells and osteosarcoma tissues.The osteosarcoma cell line U-2OS with high expression level of CKAP2 L was selected as the follow-up experiment.3.After si RNA knocks out CKAP2 L in U-2OS,its proliferation,migration and invasion ability are inhibited,and the level of apoptosis is increased.4.The expression of CKAP2 L in U-2OS/DDP is higher than that in U-2OS.5.After knocking out CKAP2 L,the proliferation,migration and invasion of U-2OS/DDP were inhibited,and its apoptosis level increased.Conclusion: This study proved that the expression of CKAP2 L in osteosarcoma cells and tissues is higher than that in normal cells and tissues.Knockout of CKAP2 L can inhibit the proliferation,migration,invasion and promote apoptosis of osteosarcoma cells.Further research found that the expression of CKAP2 L increased after the induction of U-2OS cisplatin resistance,and the knockout of CKAP2 L could inhibit the proliferation,migration and invasion of cisplatin-resistant osteosarcoma cells,and promote apoptosis.