USP11 Promotes Proliferation And Metastasis In Hepatocellular Carcinoma Via NF90

Purpose1.To explore the affection of USP11 on hepatocellular carcinoma proliferation,and metastasis.2.To explore the mechanism of USP11 on hepatocellular carcinoma proliferation,and metastasis.Methods1.In vitro,CCK-8 assay,clone formation assay were used to detect the effect of knockdown and overexpression of USP11 on proliferation of hepatocellular carcinoma cells.In vivo,the fuction of USP11 on proliferation of hepatocellular carcinoma cells was studied by establishing a subcutaneous tumorigenesis model in nude mice.2.In vitro,the relationship between USP11 and NF90 was detected by mass spectrometry,co-immunoprecipitation,western blot,and polymerase chain reaction.In subcutaneous tumors of nude mice,the protein expression of NF90 was detected by western blot assay.3.The CCK-8 assay,clone formation assay and Transwell assay were used to explore the effects of proliferation,migration and invasion ability after overexpression NF90 in hepatocellular carcinoma cells.4.The rescue experiment was performed by reintroducing NF90 into hepatocellular carcinoma cells expressing USP11-sh RNA to confirm the important fuction of NF90 in the regulation of proliferation,migration and invasion of hepatocellular carcinoma cells by USP11.5.The half-life experiment,proteasome inhibition experiment and ubiquitin detection experiment were conducted in the Control group and the USP11 knockout group to verify that USP11 regulates the expression of NF90 through deubiquitination.6.The specimen of 84 liver cancer tissues were collected and the immunohistochemical assay was used to compare the expression of USP11 and NF90.Results1.In vitro,the proliferation ability of hepatocellular carcinoma cells was decreased when USP11 knockdown.Conversely,it was increased when USP11 overexpressed.2.In vivo,the proliferation of hepatocellular carcinoma was also decreased after USP11 knockout in xenotransplantation experiments.3.The Co-IP experiments detected that USP11 and NF90 bind to each other.When USP11 knockout,the expression of NF90 was decreased in hepatocellular carcinoma cells.Conversely,the expression of NF90 was elevated when USP11 overexpressed.In addition,the expression of NF90 was also decreased in animal cancer tissues which formed by USP11 knockdown cells.4.The CCK-8 assay,clone formation assay and Transwell assay showed that NF90 overexpressed significantly promoted the proliferation,migration and invasion ability of hepatocellular carcinoma cells.5.Overexpression of NF90 can significantly rescue the decreased of proliferation,migration and invasion ability caused by USP11 knockdown.6.USP11 extended the half-life of NF90 in hepatocellular carcinomar cells and NF90 was degraded by ubiquitin-proteasome pathway.7.Statistical analysis indicated a positive correlation between USP11 and NF90 in clinical samples of hepatocellular carcinoma.Conclusion1.USP11 promotes the proliferation of hepatocellular carcinomar cells.2.NF90 promotes the proliferation and metastasis of hepatocellular carcinomar cells.3.USP11 promoting proliferation and metastasis of hepatocellular carcinoma depends on NF90.4.USP11 and NF90 can combine with each other and The former can regulate ubiquitination degradation of the latter.5.The expression of NF90 are positively associated with USP11 in clinical samples of liver cancer.