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ObjectiveTo investigate the expression of RNA binding motif single stranded interacting protein 3 （RBMS3） in epithelial ovarian cancer （EOC） tissues and its relationship with the clinicopathological features and prognosis of EOC. MethodsThe study enrolled the paraffin-embedded tissues from 110 EOC cases and 73 benign epithelial ovarian tumor cases pathologically diagnosed in the first affiliated Hospital of Bengbu Medical College from January 2015 to December 2019. By using anti-RBMS3 polyclonal antibody， the immunohistochemical staining was employed to detect RBMS3 expression in the tissues and then its correlation with the clinicopathological parameters and prognosis of EOC was analyzed. ResultsRBMS3 was expressed in both EOC and benign epithelial ovarian tumor tissues. RBMS3 expression in EOC tissues， significantly related with International Federation of Gynecology and Obstetrics （FIGO） stage， histological grade， CEA levels and survival status， was significantly lower than that in benign epithelial ovarian tumor tissues （P<0.05）. Kaplan–Meier survival curve showed that low RBMS3 expression in EOC patients was correlated with decreased progression-free survival （PFS） and overall survival （OS） （P<0.05）. Univariate analysis showed that RBMS3 expression， FIGO stage， residual lesion size， intestinal metastasis and intraperitoneal implantation were associated with OS of EOC patients （P<0.05）； multivariate analysis showed that low RBMS3 expression and intestinal metastasis were independent risk factors for poor prognosis in EOC patients （P<0.05）. ConclusionsRBMS3 is expressed at low levels in EOC tissues， which is closely related to poor prognosis of EOC patients. RBMS3 may function as a tumor suppressor gene in EOC tissues and can be used as an EOC-independent prognostic marker for targeted therapy against EOC.
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Objective:To explore the prognostic biomarkers of glioblastoma (GBM) in the tumor microenvironment (TME) and its function.Methods:A total of 169 GBM samples of 161 GBM patients were collected from the Cancer Genome Atlas (TCGA) database. ESTIMATE algorithm in R4.1.0 software was used to calculate the proportion of immune components and stromal components in TME, which were expressed as immune score and stromal score, respectively. According to the median value of the two scores, 169 GBM samples were divided into the high score group and the low score group, respectively, 84 each in each group (those whose scores were equal to the median were not involved in the grouping). The differentially expressed genes (DEG) [false discovery rate (FDR) < 0.05] between the high score group and the low score group of the two scores were obtained by using limma package, and the co-up-regulated and co-down-regulated DEG of the two scores were obtained by using Venn program. Based on the STRING database, the protein interaction (PPI) network of co-up-regulated and down-regulated DEG of immune score and stromal score was constructed, and the top 30 genes with connectivity were selected. Univariate Cox proportional hazard model analysis of overall survival (OS) of 161 GBM patients in the TCGA database was performed on co-up-regulated and down-regulated DEG between immune score and stromal score by using R4.1.0 software to obtain the DEG affecting OS. The intersection of the DEG obtained from PPI analysis and Cox analysis was taken as the prognostic core genes. According to the median expression value of prognostic core genes in GBM samples from the TCGA database, 161 patients were divided into prognostic core genes high expression group and low expression group (patients whose scores were equal to the median were not involved in the grouping), with 80 cases in each group. Kaplan-Meier survival analysis of OS was performed by using R4.1.0 software. GSEA 4.2.1 software was used to perform gene set enrichment analysis (GSEA) on all genes with transcriptome data of GBM patients in the two groups of the TCGA databases, and the main enriched functions of the two groups of genes were obtained. The CIBERSORT algorithm was used to test the accuracy of the proportion of tumor infiltrating immune cell (TIC) subsets in 169 GBM samples from the TCGA database, and 57 GBM samples were finally obtained. Immune cells with differential expression levels and immune cells related to the expression of prognostic core genes among the samples with different expression levels of prognostic core genes were analyzed; Venn program was used to obtain the intersection of immune cells with differential levels and related immune cells, and differentially expressed TIC related to expressions of prognostic core genes in GBM were obtained.Results:Based on the immune score and stromal score of GBM samples in the TCGA database, a total of 693 co-up-regulated and co-down-regulated DEG of both scores were screened out. After the intersection of 78 DEG related to OS obtained by univariate Cox regression analysis and 30 DEG obtained by PPI network results, CC motif chemokine receptor 2 (CCR2) was identified as the prognostic core gene ( HR = 1.294, 95% CI 1.060-1.579, P = 0.011). GBM patients with CCR2 high expression had worse OS compared with those with CCR2 low expression ( P = 0.009). GSEA analysis showed that genes in the CCR2 high expression group were mainly enriched in immune-related pathways, while genes in the CCR2 low expression group were mainly enriched in metabolism-related pathways. Among 57 screened GBM samples, there were differences in the levels of 3 immune cells between the CCR2 high expression group and the CCR2 low expression group ( P < 0.05). CCR2 expression was correlated with the levels of 9 immune cells (all P < 0.05). Venn program analysis showed that differentially expressed 3 TIC in GBM related to CCR2 gene expression were obtained; among them, M2 macrophages were positively correlated with CCR2 expression, while T follicular helper cell and activated NK cells were negatively correlated with CCR2 expression. Conclusions:CCR2 may be the core gene related to the prognosis in the TME of GBM. As reference, the level of CCR2 can help to predict the status of TME and prognosis in GBM patients, which is expected to provide a new direction for the treatment of GBM.
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MAGED4B belongs to the melanoma-associated antigen family; originally found in melanoma, it is expressed in various types of cancer, and is especially enriched in glioblastoma. However, the functional role and molecular mechanisms of MAGED4B in glioma are still unclear. In this study, we found that the MAGED4B level was higher in glioma tissue than that in non-cancer tissue, and the level was positively correlated with glioma grade, tumor diameter, Ki-67 level, and patient age. The patients with higher levels had a worse prognosis than those with lower MAGED4B levels. In glioma cells, MAGED4B overexpression promoted proliferation, invasion, and migration, as well as decreasing apoptosis and the chemosensitivity to cisplatin and temozolomide. On the contrary, MAGED4B knockdown in glioma cells inhibited proliferation, invasion, and migration, as well as increasing apoptosis and the chemosensitivity to cisplatin and temozolomide. MAGED4B knockdown also inhibited the growth of gliomas implanted into the rat brain. The interaction between MAGED4B and tripartite motif-containing 27 (TRIM27) in glioma cells was detected by co-immunoprecipitation assay, which showed that MAGED4B was co-localized with TRIM27. In addition, MAGED4B overexpression down-regulated the TRIM27 protein level, and this was blocked by carbobenzoxyl-L-leucyl-L-leucyl-L-leucine (MG132), an inhibitor of the proteasome. On the contrary, MAGED4B knockdown up-regulated the TRIM27 level. Furthermore, MAGED4B overexpression increased TRIM27 ubiquitination in the presence of MG132. Accordingly, MAGED4B down-regulated the protein levels of genes downstream of ubiquitin-specific protease 7 (USP7) involved in the tumor necrosis factor-alpha (TNF-α)-induced apoptotic pathway. These findings indicate that MAGED4B promotes glioma growth via a TRIM27/USP7/receptor-interacting serine/threonine-protein kinase 1 (RIP1)-dependent TNF-α-induced apoptotic pathway, which suggests that MAGED4B is a potential target for glioma diagnosis and treatment.
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Humans , Tumor Necrosis Factor-alpha , DNA-Binding Proteins/metabolism , Ubiquitin-Specific Peptidase 7 , Cisplatin , Temozolomide , Transcription Factors , Glioma , Cell Proliferation , Melanoma , Cell Line, Tumor , Apoptosis , Nuclear Proteins/geneticsABSTRACT
@#The T-cell immunoglobulin and immunoreceptor tyrosine-based inhibitory motif domain (TIGIT) is an inhibitory receptor mainly expressed on active T-cells, or natural killer cells (NK cells) that activate negative stimulus signals in immune cells by combining with multiple ligands on the surface of target cells including tumor cells and infected cells. TIGIT plays an important regulatory role in the immune pathogenesis of tumors, viral infections and various autoimmune diseases by inhibiting the over activation of cells and the over secretion of proinflammatory cytokines. Recent researches show that TIGIT is highly expressed in T cells and NK cells of cancer patients, and is related to disease progression and poor clinical prognosis. Researchers try to enhance the activity of T cells or NK cells by blocking the binding of TIGIT and its ligand for therapeutic intervention. At present, there have been many reports about the use of anti-TIGIT monoclonal antibody treatment in different mouse tumor models leading to tumor regression, TIGIT has received extensive attention in cancer immunotherapy as a promising target for next generation cancer immunotherapy. Several clinical trials are currently evaluating the efficacy of anti-TIGIT monoclonal antibodies (mAbs) in patients with several cancers. The most advanced candidate, tiragolumab, has exhibited remarkable efficacy in programmed cell death ligand 1 (PD-L1)-positive non-small cell lung carcinoma (NSCLC) patients in phase Ⅱ clinical trials, in combination with PD-L1 blockade. However, the specific mechanism of TIGIT blockade remains to be fully elucidated.
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The human telomeric i-motif (htel-iM) is a unique higher-order DNA structure formed by the cytosine-rich (C) sequences at the end of human telomeres. Studies have shown that htel-iM plays a significant role in the transcriptional regulation of telomeric repeat-containing RNA (TERRA) ‚ maintenance of telomere function and inhibition of telomerase activity. Thus‚ it is closely related to the development and progression of many cancers and is a promising new target for cancer treatment therapy. Compared to other nucleic acid higher-order structures such as G-quadruplexes‚ the stability of htel-iM is much weaker and affected by many factors like buffer pH‚ ionic conditions and molecular crowding environments. Therefore the existence of the iM structures in near neutral physiological conditions has been uncertain for a long time. Recently‚ using in-vitro screened small molecule ligands to selectively recognize and stabilize htel-iM provides a new strategy for the exploration of the biological relevance of htel-iM. Thus it has become a research hotspot to take telomere as a cancer treatment target. However‚ so far the reported small molecule ligands selectively targeting the htel-iM are far from sufficient compared with those targeting other higher-order nucleic acid structures. In this review‚ the discovery and the characteristic of the iM structures are briefly described‚ with an emphasis on the in-vitro affecting factors of the htel-iM structure‚ the reported htel-iM ligands and its biological relevance and regulation mechanisms‚ which will be helpful in further exploration of the htel-iM structure and ligand screening in near physiological conditions‚ and understanding the mechanism and developing novel therapeutic strategies for cancer treatment targeting the htel-iM structures.
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Tripartite motif containing protein 7 (TRIM7), as a member of the E3 ubiquitin ligase TRIM family, plays an important regulatory role in immune regulation, metabolism and other physiological processes. The aberrant expression of TRIM7 is closely related to the development and progression of hepatocellular carcinoma (HCC) and it shows a complex regulatory role. However, the regulatory mechanism for the expression of TRIM7 in HCC remains unknown. In this study, multiple online databases were used to analyze the expression of TRIM7 in HCC and data indicated that TRIM7 expression was upregulated in HCC and correlated to poor prognosis. Subsequently, the transcription factor binding sites in the TRIM7 promoter region were analyzed using UCSC and JASPAR databases, and the results showed that TRIM7 promoter contains four SP1 binding sites. In this work, we demonstrated that SP1 could directly bind to its binding sites in TRIM7 promoter and positively regulate the transcriptional activity driven by the TRIM7 promoter using dual luciferase reporter experiments and the ChIP-PCR method. Moreover, our results also showed SP1 overexpression upregulated the expression of TRIM7 at both mRNA and protein levels (P<0. 01),and SP1 inhibitor, mithramycin A, could reverse the activated effect of SP1 on TRIM7 expression (P<0. 01). In conclusion, this study preliminarily reveals the regulatory mechanism of TRIM7 upregulation in HCC, which provides an important theoretical basis for further study of the gene function, early diagnosis and targeted therapy.
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Tripartite motif-containing protein 28 is a kind of macromolecular protein with E3 ubiquitin ligase, which belongs to an important member of the TRIM protein family. As a new molecular biomarker, it has attracted wide attention. TRIM28 is highly expressed in many kinds of malignant tumors, which is closely related to clinicopathological features, and is also involved in biological behaviors such as proliferation, apoptosis, migration and invasion of tumor cells. TRIM28 may be a potential marker and therapeutic target for clinical diagnosis and prognosis of tumors. This study reviews the structure and biological function of TRIM28, its relationship with malignant tumors and the molecular mechanism of signal transduction pathway.
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Objective:To observe the expression of microRNA (miRNA)-7159-5p in gastric cancer tissues, and explore its effect on the proliferation and invasion of gastric cancer cells and its molecular mechanism.Methods:The cancer tissues and adjacent tissues of 29 patients with gastric cancer who underwent surgical resection in Hubei 672 Orthopedic Hospital of Integrated Traditional Chinese and Western Medicine from March 2018 to November 2019 were collected. Fluorescence real-time quantitative polymerase chain reaction (qRT-PCR) was used to detect the expression of miR-7159-5p in gastric cancer tissues and adjacent tissues, gastric cancer cell lines and normal gastric epithelial cell lines. Select the cell line with the lowest expression of miR-7159-5p, set the control group and the experimental group, and be transfected with control and miR-7159-5p mimics respectively. qRT-PCR was used to detect the expression of miR-7159-5p in transfected cells. The lymphocyte proliferation test (MTS method) was used to detect the proliferation of transfected cells, and the Transwell chamber method was used to detect the invasion activity of transfected cells. The miRNAMap database and dual luciferase reporter gene experiment were used to predict and verify the target genes of miR-7159-5p. qRT-PCR and Western blot were used to detect the expression of target genes in transfected cells.Results:The expression of miR-7159-5p in gastric cancer tissues was lower than that in adjacent tissues ( P<0.01), the expression of miR-7159-5p in gastric cancer cell lines was lower than that of normal gastric epithelial cells (all P<0.01), and the expression of miR-7159-5p was the lowest in SGC7901 cells ( P<0.01). After transfection, the expression of miR-7159-5p in SGC7901 cells of the experimental group was significantly higher than that in the control group ( P<0.01). After transfection, the proliferation activity and the cell invasion activity of the experimental group was significantly lower than that of the control group (all P<0.01). The target gene of miR-7159-5p was tripartite motif 26 (TRIM26). After transfection, the expression of TRIM26 mRNA in SGC7901 cells of the experimental group was significantly lower than that in the control group ( P<0.01). Western blot showed that after transfection, the expression of TRIM26, c-Myc, cyclin D1, β-catenin protein were lower than those in the control group (all P<0.05). Conclusions:The expression of miR-7159-5p is low in gastric cancer tissues, and miR-7159-5p can inhibit the proliferation and invasion of SGC7901 cells by down-regulating the expression of TRIM26.
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Metabolic homeostasis requires dynamic catabolic and anabolic processes. Autophagy, an intracellular lysosomal degradative pathway, can rewire cellular metabolism linking catabolic to anabolic processes and thus sustain homeostasis. This is especially relevant in the liver, a key metabolic organ that governs body energy metabolism. Autophagy's role in hepatic energy regulation has just begun to emerge and autophagy seems to have a much broader impact than what has been appreciated in the field. Though classically known for selective or bulk degradation of cellular components or energy-dense macromolecules, emerging evidence indicates autophagy selectively regulates various signaling proteins to directly impact the expression levels of metabolic enzymes or their upstream regulators. Hence, we review three specific mechanisms by which autophagy can regulate metabolism: A) nutrient regeneration, B) quality control of organelles, and C) signaling protein regulation. The plasticity of the autophagic function is unraveling a new therapeutic approach. Thus, we will also discuss the potential translation of promising preclinical data on autophagy modulation into therapeutic strategies that can be used in the clinic to treat common metabolic disorders.
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The application of clustered regularly interspaced short palindromic repeats (CRISPR) and CRISPR-associated proteins (Cas) can be limited due to a lack of compatible protospacer adjacent motif (PAM) sequences in the DNA regions of interest. Recently, SpRY, a variant of Streptococcus pyogenes Cas9 (SpCas9), was reported, which nearly completely fulfils the PAM requirement. Meanwhile, PAMs for SpRY have not been well addressed. In our previous study, we developed the PAM Definition by Observable Sequence Excision (PAM-DOSE) and green fluorescent protein (GFP)-reporter systems to study PAMs in human cells. Herein, we endeavored to identify the PAMs of SpRY with these two methods. The results indicated that 5'-NRN-3', 5'-NTA-3', and 5'-NCK-3' could be considered as canonical PAMs. 5'-NCA-3' and 5'-NTK-3' may serve as non-priority PAMs. At the same time, PAM of 5'-NYC-3' is not recommended for human cells. These findings provide further insights into the application of SpRY for human genome editing.
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Humans , CRISPR-Associated Protein 9/metabolism , CRISPR-Cas Systems , DNA , Gene Editing/methods , Streptococcus pyogenes/metabolismABSTRACT
Pulmonary fibrosis is an irreversible interstitial lung disease characterized by lung parenchyma remodeling and collagen deposition. In recent years, the incidence and mortality of pulmonary fibrosis caused by unknown causes have risen. However, its pathogenesis is still unclear. C-X-C motif chemokine ligand 12 (CXCL12)/C-X-C chemokine receptor 4 (CXCR4)/CXCR7 signal axis plays a critical regulatory role in pulmonary fibrosis disease. In addition, the signal axis has been shown to regulate recruitment and migration of circulating fibrocytes, mesenchymal stem cells to the damage lung tissue, the migration of endothelial cells, the proliferation and differentiation of fibroblasts and endothelial cells, which further affects the occurrence and progression of pulmonary fibrosis. In this review, we summarized the pathogenesis and treatment research progress of CXCL12 and its receptor CXCR4/CXCR7 in the occurrence and progression of pulmonary fibrosis.
Subject(s)
Humans , Chemokine CXCL12 , Endothelial Cells/pathology , Ligands , Lung/pathology , Pulmonary Fibrosis/pathology , Receptors, CXCR4ABSTRACT
Tripartite motif-containing protein 15 (TRIM15) is a member of the TRIM family, which is a class of proteins with E3 ubiquitin ligase activity. The function of TRIM15 in tumors is rarely reported. This study is intended to explain the role of TRIM15 in hepatocellular carcinoma. Nuclear and cytoplasmic fractionation and immunofluorescence assays confirmed that TRIM15 was located in the nucleus and cytoplasm. We designed hairpin RNA (shRNA) to knockdown TRIM15 in hepatocarcinoma cell lines. After knocking down TRIM15, cell growth curve and clone formation assays showed that cell proliferation was significantly inhibited (P<0. 05). Cell cycle analysis by flow cytometry showed that knockdown of TRIM15 blocked cell cycle in the G
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Metastasis and cell infiltration are the difficulties in the treatment of solid and lymphatic carcinoma and the main causes of disease recurrence and death. The migration of cancer cells is a prerequisite for tumor metastasis and invasion. The CXCL12-CXCR4 pathway plays an important role in the pathogenesis of solid tumors and leukemia. The interaction between CXCL12 and its receptor CXCR4 can activate multiple signaling pathways and regulate different physiological and pathophysiological processes. Thus, blocking CXCL12-CXCR4 binding and / or downstream pathways has clinical benefits in treating a variety of diseases and cancers. Although some CXCL12 and CXCR4 antagonists have been identified and have shown encouraging results in terms of antitumor activity, these drugs have not been widely used in clinical patients due to their serious toxic and side effects. There is an urgent need to develop novel CXCL12-CXCR4 axis antagonists for the treatment of tumors. Herein, we review the recent research progress of CXCR4 pathway in solid tumors and leukemia, and discuss the therapeutic value and future research direction of CXCR4 pathways in solid tumors and leukemia.
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Objective To explore the expression and significance of CC mokif chemokine ligand 20(CCL20) gene in esophageal carcinoma and its influence on the progression and prognosis of the disease. Methods The expression data of esophageal cancer were obtained from the Cancer Genome Atlas (TCGA) database, and were collected from cBioPortal(www.cbioportal.org).The survival curve was analyzed by Kaplan-Meier method. Through the website (http://ualcan.path.uab. edu) the objective was to analyze the expression of CCL20 gene in esophageal cancer tissues, the relationship between CCL20 and obesity and the methylation level of CCL20 in esophageal cancer. cBioPortal was used to analyze the copy number and mRNA expression of CCL20 gene. Finally, the expression differences of CCL20 in esophageal cancer tissues and adjacent tissues were analyzed by mRNA and protein levels. Results CCL20 was highly expressed in esophageal cancer, especially adenocarcinoma. The survival rate of patients with high expression of CCL20 was reduced. The expression of CCL20 was directly proportional to the degree of obesity in patients with esophageal cancer and inversely proportional to the degree of methylation. The expression of CCL20 in esophageal cancer tissues increased significantly. Conclusion CCL20 gene expression level is associated with poor prognosis of esophageal cancer patients.
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Objective:To explore the possible mechanism of Astragali Radix-Curcumae Rhizoma （AC） in inhibiting tumor growth in the orthotopic transplantation model of colon cancer in mice. Method:The molecular docking technology was used to predict the intermolecular interaction between the main active components of AC and the pathway target proteins， such as stromal cell-derived factor-1 （SDF-1）， C-X-C motif chemokine receptor 4 （CXCR4）， and nuclear factor kappa-B p65 （NF-<italic>κ</italic>B p65）. The orthotopic transplantation model of CT26.WT colon cancer was established in mice for <italic>in vivo</italic> experimental verification. Sixty BALB/c male mice were randomly divided into a sham operation group， a model group， a 5-fluorouracil （5-Fu， 30 mg·kg<sup>-1</sup>） group，and low- （0.32 g·kg<sup>-1</sup>）， medium- （0.64 g·kg<sup>-1</sup>）， and high-dose （1.28 g·kg<sup>-1</sup>） AC groups， with 10 mice in each group. The sham operation group and the model group received normal saline by gavage. The corresponding drugs were administered by gavage in the 5-Fu group and by intraperitoneal injection in the AC groups. After intervention for 15 days， the tumor <italic>in situ</italic> was completely stripped， and the colon tissues 5-6 cm in length adjacent to the tumor were taken. The tumor volume was measured and calculated. The pathological changes of tumor tissues and colon tissues were observed by Hematoxylin-Eosin （HE） staining. Western blot was used to detect the protein expression of SDF-1， CXCR4， p-NF-<italic>κ</italic>B p65 in colon tissues. Western blot and Real-time quantitative polymerase chain reaction （Real-time PCR） were used to detect SDF-1， CXCR4， NF-<italic>κ</italic>B p65， Cyclin D<sub>1</sub>， oncogene c-Myc protein and mRNA expression in tumor tissues. Result:Compared with the model group， 5-Fu and AC groups showed reduced tumor volumes <italic>in situ</italic> （<italic>P</italic><0.05， <italic>P</italic><0.01）， with the tumor inhibition rate in the 5-Fu group as high as （61.38±2.34）%. The tumor-inhibiting effect was optimal in the medium-dose AC group， with the tumor inhibition rate of （43.43±3.71）%. Compared with the model group， 5-Fu and AC groups showed relieved pathological changes of tumor and colon tissues. Specifically， AC down-regulated the protein expression levels of SDF-1， CXCR4， and p-NF-<italic>κ</italic>B p65 in colon tissues （<italic>P</italic><0.01）， and down-regulated the protein and mRNA expression levels of SDF-1， CXCR4， NF-<italic>κ</italic>B p65， Cyclin D<sub>1</sub>， and c-Myc in tumor tissues （<italic>P</italic><0.05， <italic>P</italic><0.01）. Conclusion:AC can inhibit the growth of orthotopic transplantation tumor of colon cancer， and its intervention mechanism may be related to the regulation of related protein and mRNA expression in the SDF-1/CXCR4/NF-<italic>κ</italic>B signaling pathway.
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The functionality of DNA biomacromolecules has been widely excavated, as therapeutic drugs, carriers, and functionalized modification derivatives. In this study, we developed a series of DNA tetrahedron nanocages (Td),
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E3 ubiquitin ligase TRIM21(tripartite motif containing 21) plays an important role in regulating cell biological functions and clinical prognosis as oncogene or tumor suppressor in different types of tumors. However,the biological functions and molecular mechanism of TRIM21 in hypopharyngeal squamous cell carcinoma (HPSCC) are still unclear.Our results showed that TRIM21 is highly expressed in moderately and well differentiated HPSCC, suggesting the role of TRIM21 in tumor differentiation. Overexpression and knockdown of TRIM21 inhibited or promoted cell proliferation and migration. Meanwhile, the expression of differentiation markers including KRT10 (keratin 10), IVL (involucrin) and TGM1 (transglutaminase 1) were increased and decreased upon TRIM21 overexpression or knockdown, respectively. The bioinformatics analysis of TRIM21 interacting protein identified by co-immunoprecipitation combined with mass spectrometry suggested that TRIM21 may be closely related to the regulation of cytoskeleton protein. We further demonstrated that TRIM21 interacted with KRT10. The inhibition of protein synthesis by cycloheximide led to upregulation of KRT10 in TRIM21 overexpressing FaDu cells, and ectopic expression of TRIM21 enhanced the ubiquitination level of KRT10. In summary, our results suggest that TRIM21 may promote the HPSCC differentiation by mediating ubiquitination of cytoskeleton proteins to improve the protein stability.
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Autophagy is a common cellular metabolic process, which is characterized by the formation of double membrane structures named autophagosomes to degrade intracellular components or invading foreign substances to maintain cellular homeostasis. Autophagy is crucial for maintaining cell homeostasis. The dysfunction of autophagy is closely related to the occurrence and development of various diseases, including tumors, neurodegenerative diseases, viral infection, immune diseases and so on. Autophagy may be a potential therapeutic target for these diseases. Therefore, the investigation of autophagy regulation is a hot issue in life science and medical research. The TRIM (tripartite motif-containing proteins) family is a set of proteins with E3 ubiquitin ligase activity and usually contains three conserved domains, a RING zinc finger structure, a B-box structure and a coiled helix domain. Many TRIM family members have been found to play important roles in autophagy regulation, the mechanism of which include modulating autophagy-related signaling pathways, regulating autophagy core molecules and acting as autophagy receptors, etc. TRIMs participate in many biological pathways through regulating autophagy, such as immunity, virus infection and tumors. This review covers the role of TRIM proteins in regulating autophagy, the molecular mechanism and the corresponding biological effects.
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Objective To explore the effect of C-X-C motif chemokine ligand-13 (CXCL-13) on the proliferation and migration of human bone marrow mesenchymal stem cells (BMSCs) by network pharmacology. Methods To predict that the targets of CXCL-13 on BMSCs by online database. Metascape was used to perform gene ontology (GO) of the targets and Kyoto encyclopedia of genes and genomes (KEGG) pathway was used to perform enrichment analysis. The protein interaction analysis was performed by STRING 11.0 database, and the protein module of core gene was screened by using the cytoHubba 0. 1 of Cytoscape 3. 8. We divided BMSCs into control group, CXCL-13 group and PI3K inhibitor group. MTT assay, flow cytometric analysis and Transwell cell migration assay were respectively used to detect the absorbance (A) value of BMSCs in each group, the apoptosis rate and the number of cell migration. The protein contents of epidermal growth factor (EGF) and vascular endothelial growth factor (VEGF) in BMSCs supernatant were determined by ELISA. Western blotting was used to detect the protein expression of Akt and phosphorylated Akt (p-Akt) of BMSCs in each group. Results It was predicted that 21 targets of CXCL-13 effect on BMSCs. There were 32 biological processes related to cell proliferation include stem cell proliferation, regulation of endothelial cell proliferation and positive regulation of smooth muscle cell proliferation. There were 22 biological processes related to cell migration include regulating cell migration, amebic cell migration and endothelial cell migration. There were 40 KEGG pathways including cancer pathway, PI3K-Akt signaling pathway and MAPK signaling pathway. The core proteins included tumor protein P53 (TP53), epidermal growth factor receptor (EGFR), heat shock protein 90 kD alpha class B member 1 (HSP90AB1), protein kinase Ca (PRKCA), estrogen receptor 2 (ESR2) and prostaglandin E receptor 4 (PTGER4). Compared with other groups, the absorbance (A) value and cell migration number of BMSCs in CXCL-13 group increased significantly (P< 0. 01, 71=15), and the apoptosis rate decreased significantly (P<0. 01, n= 15). However, absorbance value, apoptosis rate and migration number of BMSCs in PI3K inhibitor group were contrary to those in CXCL-13 group (P<0. 01, n= 15). Compared with the control group, the protein contents of EGF and VEGF in BMSCs of CXCL-13 group increased significantly (P<0. 01, n= 15), and the relative expression of Akt and p-Akt increased significantly (P<0. 01, n = 9). However, the protein content of EGF and VEGF, and the relative expression of Akt and p-Akt in PI3K inhibitor group were opposite. Conclusion Through activating PI3K-Akt pathway, CXCL-13 may promote BMSCs paracrine EGF and VEGF proteins, and improve proliferation and migration of BMSCs, as well as inhibit BMSCs apoptosis.
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AIM: To study the correlation between the expression of recepteur d'origine nantais (RON) protein and CXC chemokine motif receptor 4 (CXCR4) protein and abiraterone resistance in patients with castration-resistant prostate cancer (CRPC). METHODS: From January 2017 to February 2020, 127 patients with CRPC who were treated with abiraterone in our hospital were selected. According to the status of drug resistance, they were divided into observation group (n=32, patients with abiraterone resistance) and control group (n=95, patients in remission). Immunohistochemistry and Western blot analysis were used to compare the expression of RON and CXCR4 protein between the two groups. Logistic regression analysis was used to conduct single-factor and multi-factor analysis of RON, CXCR4 protein and drug resistance, and receiver operating characteristic curve (ROC) and area under ROC (AUC) were used to analyze the value of RON and CXCR4 protein in predicting drug resistance. In addition, RON and CXCR4 inhibitors were added to abiraterone-resistant cell lines. The effects of the two on the apoptosis indicators of abiraterone resistance[caspase-3, caspase-9, apoptosis rate] were observed. RESULTS: Immunohistochemistry showed that the positive expression rate of RON in the observation group (71.88%, 23/32) was higher than that of the control group (27.37%, 26/95). The positive expression rate of CXCR4 in the observation group (65.63%, 21/32) was higher than that of the control group (12.63%, 12/95). Western blot detection showed that the RON and CXCR4 proteins in the observation group were higher than those in the control group (P 4.11, the sensitivity was 84.37%, and the specificity was 61.05% (P 3.42, the sensitivity was 75.00%, and the specificity was 80.00% (P< 0.05). The AUC of RON+CXCR4 protein predicting abiraterone resistance was 0.884 (95%CI: 0.815-0.934), the sensitivity was 87.50%, and the specificity was 83.16% (P< 0.05). CONCLUSION: The expression of RON and CXCR4 protein in CRPC patients increases significantly, which is closely related to the resistance of Abiraterone in patients and is expected to become a marker for predicting drug resistance. Inhibiting the expression of RON and CXCR4 proteins can promote the apoptosis of CRPC abiraterone resistant cells.