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Objective:To investigate the effect of ubiquitin binding enzyme 2T (UBE2T) on the radiosensitivity of lung adenocarcinoma and unravel its possible mechanism.Methods:A total of 45 patients pathologically diagnosed with different stages of lung adenocarcinoma and treated with radiotherapy in the Second Affiliated Hospital of Zunyi Medical University from March, 2019 to December, 2021 were enrolled, and the efficacy was evaluated according to response evaluation criteria in solid tumors (RECIST1.1). All patients were divided into radiosensitive group ( n=25) and radioresistant group ( n=20). Radiosensitive group was complete remission (CR)+partial remission (PR), and radioresistant group was stable disease (SD) + progression disease (PD). Immunohistochemistry (IHC) was used to calculate the score based on the staining intensity and the number of positive cells. Chi-square test was combined to analyze the correlation between the expression level of UBE2T in paraffin specimens of lung adenocarcinoma patients and the radiosensitivity of patients. Lentivirus UBE2T-interfered (UBE2Tsh) A549 and UBE2T-overexpressed SPC-A-1 lung adenocarcinoma cells and their respective controls were constructed for irradiation and colony formation assay. The survivor fraction curve was fitted by single-hit multi-target model. The DNA double-strand break (DSB) marker γH2AX foci were detected by immunofluorescence (IF). The expression levels of UBE2T, γH 2AX and Rad51 proteins were detected by Western blot. Cell cycle and apoptosis rate of A549 were determined by flow cytometry. Binary variables were statistically analyzed by Fisher's exact probability method and measurement data were assessed by t-test. Results:High-expression level of UBE2T was correlated with the radiosensitivity of lung adenocarcinoma patients ( P<0.05). UBE2Tsh improved the radiosensitivity of A549 lung adenocarcinoma cells, and the sensitizing enhancement ratio (SER) was 1.795. UBE2T overexpression decreased the radiosensitivity of SPC-A-1 lung adenocarcinoma cells with an SER of 0.293. γH2AX foci number per cell were significantly increased in UBE2Tsh A549 cells after irradiation ( P<0.01) . Compared with the control group, the expression level of γH2AX protein was up-regulated ( P<0.01)and that of Rad51 protein was down-regulated in UBE2Tsh A549 cells after radiation ( P<0.001). Compared with the control group, the expression level of γH2AX protein was down-regulated ( P<0.05) and that of Rad51 protein was up-regulated in UBE2T overexpressed SPC-A-1 cells ( P<0.001). The proportion of UBE2Tsh A549 cells in G 2 phase was decreased ( P<0.01) and cell apoptosis was increased ( P<0.001). Conclusions:UBE2T might promote the radioresistance of lung adenocarcinoma cells by enhancing DNA DSB repair induced by radiotherapy, inducing cell cycle G 2 phase arrest, and reducing cell apoptosis.
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Objective: To investigate the expression and clinical significances of ubiquitin conjugating enzyme E2 T (UBE2T) in hepatocellular carcinoma (HCC), and explore the role of UBE2T gene expression in migration and invasion of HCC cells. Methods: The expression of UBE2T mRNA in 54 pairs of HCC tissues and the matched noncancerous liver tissues was analyzed by real-time fluorescent quantitative PCR. The expression level of UBE2T protein in 233 cases of HCC tissues was detected by immunohistochemistry, then the relationship between UBE2T expression and clinicopathological features of HCC patients was analyzed. The recombinant lentiviral vector pWPXL-UBE2T contained UBE2T gene sequences was constructed and further transfected into Hep3B and SMMC-7721 cells. In the other hand, the recombinant lentiviral vector GV248-shUBE2T-1 and GV248-shUBE2T-2 targeted UBE2T gene were transfected into MHCC-LM3 and SK-Hep1 cells. The forced expression and knockdown of UBE2T gene were validated by real-time fluorescent quantitative PCR and Western blotting. Afterwards, the effects of UBE2T gene overexpression and silencing on migratory and invasive abilities of HCC cells were detected by Transwell chamber assays, respectively. Results: The UBE2T mRNA level in HCC tissues was significantly higher than that in the matched noncancerous liver tissues (P < 0.000 1). The expression of UBE2T protein was correlated with intrahepatic metastasis of HCC (P = 0.004) in 233 patients with HCC. Compared with the control group, the forced expression of UBE2T gene markedly promoted both migration and invasion of Hep3B and SMMC-7721 cells (both P < 0.01); while the knockdown of UBE2T gene obviously inhibited the migration and invasion of MHCC-LM3 and SK-Hep1 cells as compared with the negative control group (both P < 0.05). Conclusion: High expression level of UBE2T in HCC tissues is correlated with the presence of intrahepatic metastasis, and the overexpression of UBE2T gene can promote the migration and invasion of HCC cells.
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UbcH5c belongs to the ubiquitin-conjugating enzyme family and plays an important role in catalyzing ubiquitination during TNF---triggered NF-B activation. Therefore, UbcH5c is a potent therapeutic target for the treatment of inflammatory and autoimmune diseases induced by aberrant activation of NF-B. In this study, we established a stable expression system for recombinant UbcH5c and solved the crystal structure of UbcH5c belonging to space group P222with one molecule in the asymmetric unit. This study provides the basis for further study of UbcH5c including the design of UbcH5c inhibitors.
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BACKGROUND: The prevalence of novel type 1 diabetes mellitus (T1DM) antibodies targeting eukaryote translation elongation factor 1 alpha 1 autoantibody (EEF1A1-AAb) and ubiquitin-conjugating enzyme 2L3 autoantibody (UBE2L3-AAb) has been shown to be negatively correlated with age in T1DM subjects. Therefore, we aimed to investigate whether age affects the levels of these two antibodies in nondiabetic subjects. METHODS: EEF1A1-AAb and UBE2L3-AAb levels in nondiabetic control subjects (n=150) and T1DM subjects (n=101) in various ranges of age (18 to 69 years) were measured using an enzyme-linked immunosorbent assay. The cutoff point for the presence of each autoantibody was determined based on control subjects using the formula: [mean absorbance+3×standard deviation]. RESULTS: In nondiabetic subjects, there were no significant correlations between age and EEF1A1-AAb and UBE2L3-AAb levels. However, there was wide variation in EEF1A1-AAb and UBE2L3-AAb levels among control subjects <40 years old; the prevalence of both EEF1A1-AAb and UBE2L3-AAb in these subjects was 4.4%. When using cutoff points determined from the control subjects <40 years old, the prevalence of both autoantibodies in T1DM subjects was decreased (EEFA1-AAb, 15.8% to 8.9%; UBE2L3-AAb, 10.9% to 7.9%) when compared to the prevalence using the cutoff derived from the totals for control subjects. CONCLUSION: There was no association between age and EEF1A1-AAb or UBE2L3-AAb levels in nondiabetic subjects. However, the wide variation in EEF1A1-AAb and UBE2L3-AAb levels apparent among the control subjects <40 years old should be taken into consideration when determining the cutoff reference range for the diagnosis of T1DM.
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Humans , Young Adult , Antibodies , Autoantibodies , Diabetes Mellitus, Type 1 , Diagnosis , Enzyme-Linked Immunosorbent Assay , Eukaryota , Peptide Elongation Factor 1 , Peptide Elongation Factors , Prevalence , Reference ValuesABSTRACT
Ubiquitin-conjugating enzyme 2C (UBE2C)is involved in cell cycle progression,mitosis regulation and targeted degradation of short-lived proteins.UBE2C is the cancer-related ubiquitin-conjugating enzyme,which is significantly higher in breast cancer tissues compared to the adjacent normal tissue.The abnormal expression of UBE2C participates in the process of occurrence,development and metastasis of breast cancer by the regulation of cell cycle progression,C-erbB-2,cell apoptosis factors and metastasis related genes. It will provide reference for the diagnosis and prognosis of breast cancer.
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Ubiquitin-proteasome pathway is one of the primary pathway in intracellular protein degrada-tion,therefore the ubiquitin conjugating enzyme D3(UbE2D3)which involved in the ubiquitin mineralization process can affect the biological effects accordingly to affect some protein and nucleic acid content and activity. The function that participate in modification and degradation of some cancer factors is vital in tumor cells,and followed by tumor biological behavior changing. Researches show that UbE2D3 correlates with human telomer-ase reverse transcriptase( hTERT),radiation sensitivity,aggressive,etc in breast cancer.
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More and more evidences have shown that disorder of ubiquitination regulates the genesis and development of tumors.Ubiquitin-conjugating enzymeE2C(UBE2C), an important member of Ubiquitin-Conjugating Enzyme family, is the key factor for metaphase-metaphase transition regulation .In recent years , many studies have focused on the relationship between UBE2 C and tumors .This article reviews the progress of UBE2C expression status in tumors, its influence on genesis and development of tumors and the underlying mechanism .
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[Abstract ] Objective Bioinformatics provides a lot of valuable information for online prediction of new genes.In this study, we predicted the biological function of ubiquitin-conjugating enzyme 2S ( UBE2S) based on bioinformatics. Methods The UBE2S gene was screened and cloned from the cDNA library of human HepG2 cells.The relationship of the structure and function of UBE2S was explored based on the full-length cDNA library.MEGA5.05, CLUSTALW2 and SWISS-MODEL were used to study the phylogeny, conservation, and 3D structure of UBE2S. Results The UBE2S gene encoded a polypeptide of 241 residues with a predicted molec-ular weight of 23 770 and an isoelectric point of 8.81.The UBE2S protein contained no transmembrane locus and the probabilities of their functions of growth factors, cation channel and structural protein were 8.904, 0.313, and 0.291.The analysis of BLASTp showed that the isolated UBE2S had a 90-97%identity with the other species. Conclusion Analysis of the structure and function of the UBE2S protein can not only provide more information about its gene family but also pave the way for further experimental studies on the molecular mechanism of the consequent hepatocellular carcinoma.