| With the increasing of morbidity and mortality,cancer becomes the leading cause of death in China.Lung cancer,as the first mortality rate and the first morbidity rate of malignant cancer,is widely researched.Non-small cell lung cancer(NSCLC)has the characteristics of operation difficulty,metastasis and recurrence,and unsensitivity to chemotherapy,which are difficult for traditional treatment.The research of NSCLC targeted drugs is becoming more and more significant to therapy of NSCLC,and the treatment of NSCLC is gradually explored through targeting deubiquiting enzymes.There are 5 families of DUB,of which USP is the largest.USP 13,a popular member of the USP family,was extensively studied in functional regulation from 2016 to 2018,and published 5 paper in Natrue Communication.Its main functions are to enhance the immunity against infection,to regulate DNA damage repair,to amplify abnormal expression and to regulate metabolic regulation of ovarian cancer,to control endoplasmic Reticulum related degradation,to regulate and control oncogene in carcinogenesis.Objective: This research analyzes the expression of m RNA of USP13 in normal tissues and tumors from the perspective of abnormal amplification of USP13 in different cancer,and explores the reason of abnormal amplification of USP13 in NSCLC.To analyze the relationship between the amplification and expression of USP13 gene in NSCLC by TCGA(The Cancer Genome Atlas)Database,to screen out proteins that may have potential relationship with USP13,and to explore the mechanisms of USP13 in NSCLC in vitro and in vivo.Methods: Through TCGA database,we explored the changes of genome-related CNV by the percentage of variation in the number of cancerous patient.The level of m RNA expression of USP13 in tumors and the possible associated genes of USP13 were analysed using the transformation from UCSC to a log2(RSEM + 1)as a level 3 data stored in cg Data.By means of molecular cloning,the relationship between USP 13 domain and location was investigated by using GFP fluorescence label on USP 13 in different domains.USP 13 was knocked down by lentivirus stable transfection for stabilizing gene silence(sh-RNA)to investigate the role of USP 13 in NSCLC cells.The role of USP13 in tumor formation in vivo was investigated by immunodeficient mice.Results: from the m RNA expression of USP13,it was found that the expression of m RNA in USP13 was up regulated in sarcoma,prostate and colon cancer,and down regulated in lung cancer,breast cancer,cervical cancer,liver cancer,uterine cancer,cholangiocarcinoma,thyroid cancer and kidney cancer.From the amplification of USP13 gene,the USP13 gene in the lung squamous cell carcinoma was the largest(CNV,copy number variation),and appeared the first amplification.By analyzing the amplification of TCGA lung squamous cell carcinoma and the expression of m RNA level,we found that the significant amplification of USP13 in lung squamous cell carcinoma was due to a significant amplification area of the q arm of No.3 staining,and the CNV and m RNA expression in the amplified region was positively correlated.From the protein level of clinical patients,the expression of USP13 in cancer is slightly higher than that in cancer.USP13 is located in the nucleus and cytoplasm.From the functional point of view,through the sequencing data of TCGA lung cancer patients,the genes with high correlation with USP13 were screened,and about 20 genes potentially related to USP13 were screened.The functional and structural interactions of these genes were analyzed by GO analysis,and the potential regulatory functions of USP13 were found(lung adenocarcinoma may be associated with splicing,and lung squamous cell carcinoma may be associated with phosphine).Transfecting different domains of USP13 in cells,we found that the Zn-Finger deletion structure in cells was point-like.In NSCLC cell lines,using gene silencing techniques to knock down USP13,we found that USP13 stabilize p AKT which slowed cell growth and weakened early tumorigenesis in mice. |
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