| Background:Gastric cancer is one of the most common gastrointestinal tumours in China and the world,with the highest incidence and mortality rate among all tumours.Due to the insidious onset of stomach cancer,it is difficult to achieve early detection,early diagnosis and early treatment.Most of the patients are already at an advanced stage when they are discovered,which deprives them of the best time for surgical treatment and affects their survival time.Extensive literature study revealed that USP1 is abnormally expressed in various tumor tissues,including breast cancer,osteosarcoma and hepatocellular carcinoma,and it is involved in the proliferation and migration of tumors through various signaling pathways.Although there are a few reports on the role of USP1 in gastric cancer,the molecular mechanism remains unclear.This study aimed to investigate the role and mechanism of USP1 in the proliferation and migration of gastric cancer cells and identify new targets for the treatment of gastric cancer.Objectives:1.To explore the expression and biological functions of USP1 in gastric cancer.2.To examine the potential molecular mechanism of USP1 and provide a theoretical basis for targeted therapies of gastric cancer.Methods:1.Bioinformatics analysis was performed,which involved obtaining data related to tissues and cells in TCGA and CCLE databases.Gene knockdown screening of different cell lines of gastric cancer by gene effect score..2.A plasmid vector for USP1 was constructed,and then transfected into gastric cancer cell line(AGS)using a cell counting kit(CCK-8),the migration ability and the clone formation were assessed to determine the role of USP1 in the proliferation and metastasis of gastric cancer.3.The transcriptome of knockdown USP1 gastric cancer cell line was sequenced and the relevance of USP1 to gastric cancer was revealed by differential gene expression,differential gene function and signaling pathways.4.Expression of c-JUN at mRNA and protein level in the USP1 knockdown and inhibitor groups were detected by using q PCR and Western blot.5.The rescue experiment was validated by overexpressing c-JUN in USP1 knockdown AGS gastric cancer cells and observing the effect of USP1 on proliferation and migration of gastric cancer cells.c-JUN was also overexpressed in the inhibitor group to observe the ability of USP1 on the proliferation and migration of gastric cancer cells.Results:1.Bioconductivity analysis showed that USP1 was significantly expressed in gastric cancer tissues,and screened gastric cancer cells for subsequent experiments.2.USP1 knockdown inhibited the proliferation and migration of gastric cancer cells.3.The transcriptome of USP1 knockdown cells was sequenced,and the transcriptional data were analyzed by differential gene expression,pathway analysis and clustering analysis.USP1 was found to be significantly correlated with MAPK/cJUN.4.USP1 and c-JUN mRNA and protein levels were significantly decreased in AGS with knockdown of USP1.After the addition of inhibitors,USP1 and c-JUN mRNA and protein levels were decreased further.5.The proliferation and migration ability of USP1 knockdown gastric cancer cells were reduced.However,overexpression of c-JUN in the USP1 knockdown gastric cancer cell line was able to reverse this effect.Overexpression of c-JUN in the suppressor group similarly reversed the proliferation and migration abilities.Conclusion:USP1 was associated with the proliferation and migration of gastric cancer cells,and its expression was upregulated in gastric cancer tissues.USP1 knockdown significantly inhibited the proliferation and migration of gastric cancer cells.Morever,transcriptome sequencing that USP1 interacted with MAPK/c-JUN to regulate the proliferation and migration of gastric cancer cells.USP1 may serve as a predictor of gastric cancer proliferation and migration,and may also be investigated as a potential therapeutic target for gastric cancer,which has certain translational applications and deserves further validation. |
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