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@#Objective To investigate the effects of targeted silencing of CXCL5 on the related biological functions of laryngeal squamous cell carcinoma(LSCC)cell line AMC-HN-8,and analyze the regulation through TCGA database.Methods RNA-seq data related to LSCC were obtained from TCGA database,and the expression differences of CXCL5 gene in LSCC and the adjacent tissues were analyzed. Total 60 samples of LSCC and the adjacent tissues from January 2019 to December 2020 were selected from the First Hospital of Shanxi Medical University,and the expression of CXCL5 protein in LSCC tissues was detected by immunohistochemical staining. Human LSCC cell line AMC-HN-8 was cultured and the mRNA transcription level of CXCL5 in AMC-HN-8 cells was detected by qPCR. Two groups of SiRNA with high knock-down efficiency were screened,CCK8 assay was used to detect the cell proliferation,Transwell test was used to measure the cell invasion and migration,and flow cytometry was used to detect the cell cycle and apoptosis. The correlation between CXCL5and tumor immune invasion level of LSCC was analyzed by ssGSEA,and CXCL5 co-expression gene network was constructed and analyzed for GO and KEGG enrichment.Results Compared with the adjacent tissues and the cells in control group,the expression of CXCL5 in LSCC tissues and cells increased,which was consistent with the analysis of TCGA database;Inhibition of CXCL5 expression in AMC-HN-8 cells inhibited the proliferation,invasion and migration of tumor cells,and promoted the apoptosis through inhibiting the cell cycle in G1 phase;The immune cell scores in DC,neutrophils,NK and TH17 cells were different.Conclusion CXCL5gene is highly expressed in LSCC tissues,which might be one of the targets of LSCC targeted therapy.
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ObjectiveTo investigate the effect of cSN50.1 on the proliferation， migration， invasion， and colony formation of HepG2 cells and its mechanism. MethodsHepG2 cells were divided into cSN50.1 0 μmol/L， cSN50.1 10 μmol/L， cSN50.1 30 μmol/L， cSN50.1 50 μmol/L， cSN50.1 70 μmol/L， and cSN50.1 90 μmol/L groups， and CCK－8 assay was used to investigate the effect of different concentrations of cSN50.1 on the proliferation of HepG2 cells and calculate half－maximal inhibitory concentration （IC50）. HepG2 cells were divided into cSN50.1 0 μmol/L， cSN50.1 10 μmol/L， cSN50.1 30 μmol/L， and cSN50.1 50 μmol/L groups， and wound healing assay， Transwell assay， and colony－forming assay were used to investigate the effect of different concentrations of cSN50.1 on the migration， invasion， and colony formation of HepG2 cells. HepG2 cells were divided into Control group， SP600125 group （an inhibitor of the AP－1 signaling pathway）， and cSN50.1 group to investigate the influence of the AP－1 signaling pathway on the effect of cSN50.1 on hepatocellular carcinoma cells， and RT－PCR and Western Blot were used to measure the expression of CXCL5， TNF－α， and c－Jun protein in cytoplasm and nucleus. HepG2 cells were divided into Control group， PDTC group （an inhibitor of the NF－κB signaling pathway）， and cSN50.1 group to investigate the influence of the NF－κB signaling pathway on the effect of cSN50.1 on hepatocellular carcinoma cells， and RT－PCR and Western Blot were used to measure the expression of CXCL5， TNF－α， and NF－κB protein in cytoplasm and nucleus. A one－way analysis of variance was used for comparison between multiple groups， and the SNK－q test was used for further comparison between two groups. ResultsCompared with the 0 μmol/L group， the 10 μmol/L group had no significant changes in proliferation， migration， invasion， and colony formation abilities （P >0.05）； the 30 μmol/L group had no significant change in proliferation ability （P>0.05）， but with significant reductions in migration， invasion， and colony formation abilities （P<0.05）； the 50 μmol/L group had significant reductions in proliferation， migration， invasion， and colony formation abilities （all P<0.01）； the 70 μmol/L and 90 μmol/L groups had a significant reduction in cell proliferation ability （P<0.01）， but with a cell survival rate of below 50%. Compared with the Control group， the SP600125， PDTC， and cSN50.1 groups had significant reductions in the mRNA and protein expression levels of CXCL5 and TNF－α （all P<0.05）. Compared with the Control group， the SP600125 group， the PDTC group， and the cSN50.1 group had a significant reduction in nuclear protein of c－Jun and NF－κB expression （P<0.05）； the SP600125 group and the PDTC group had a significant reduction in cytoplasmic protein of c－Jun and NF－κB expression （P<0.05）； the cSN50.1 group had a significant increase in cytoplasmic protein of c－Jun and NF－κB expression （P<0.05）. ConclusionThis study shows that cSN50.1 can inhibit the malignant behavior of hepatocellular carcinoma cells and reduce the expression of CXCL5 and TNF－α by inhibiting the nuclear import of c－Jun and NF－κB in hepatocellular carcinoma cells.
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Background and Aim: Despite the economic importance of buffalo as a main source of milk and meat, only little attention has been directed to its immune and reproductive performance. The early diagnosis of subclinical endometritis may reduce the economic loss of buffalo’s production. The difference in expression profiles of immunity-related genes has an important role in the early detection of subclinical endometritis. This study aimed to assess the expression of five immunity-related genes: TGFBR1, PTGER2, PTGER4, HP and CXCL5 in endometritis-infected buffaloes. Materials and Methods: Total RNA was extracted from 120 buffalo uterine samples; 60 infected with endometritis and 60 healthy ones. Qt-PCR was performed on cDNA synthesized from extracted RNA using Sybr green and GAPDH as a house-keeping gene. Results: The results showed the up-regulation of two tested genes; TGFBR1 and CXCL5 in endometritis-infected buffalo compared to healthy animals by 7.9 and 4.3 folds, respectively at a significance level of p<0.05. The other three tested genes; PTGER2, PTGER4 and HP were down-regulated in buffalo during endometritis infection at different levels; PTGER2 and HP (0.6 folds, p<0.05) and PTGER4 (0.4 fold, p= 0.2). Conclusions: It is to be concluded that the assessment of expression of inflammation-related immunity genes may have an effective role on the detection of endometritis infection in buffalo during its early stages and this early diagnosis can reduce the economic loss of buffalo production and reproduction.
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Objective: To explore the underlying mechanisms of drug-resistance of non-muscle invasive bladder cancer (NMIBC) to the intravesical chemotherapy. Methods: The chemoresistant bladder cancer cell line M-RT4 was established by continuous exposure of RT4 cells to mitomycin C (MMC). The chemoresistance and migration of M-RT4 cells were detected by CCK-8 assay and wound healing assay, respectively. The expressions of CXC chemokine ligand 5 (CXCL5) mRNA and protein in RT4 and M-RT4 cells as well as the recurrent NMIBC tissues were detected by real-time fluorescent quantitative PCR and Western blotting, respectively. The sensitivity of RT4 cells to MMC was observed after the treatment of recombinant human CXCL5 (rhCXCL5). The change of resistance of M-RT4 cells to MMC was detected by CCK-8 assay after siRNA transfection for CXCL 5 gene knockdown. The expressions of epithelial-mesenchymal transition (EMT)-associated factors [E-cadherin, cytokeratin-7 (Cyt-7), Claudin-1, N-cadherin, Vimentin and Snail] and nuclear factor-kappa B (NF-κB)-associated factors (NF-κB1, Rel A, NF-κB2 and Rel B) during the drug resistance of NMIBC cells were detected by Western blotting. Results: Compared to RT4 cells, the resistance of M-RT4 cells to MMC was significantly increased (P < 0.01), while the proliferation and migration of M-RT4 cells were enhanced (P < 0.05, P < 0.01). CXCL5 mRNA was over-expressed in the recurrent NMIBC tissues (P < 0.000 1), while the expression levels of CXCL5 mRNA and protein in M-RT4 cells were higher than those in RT4 cells (both P < 0.01). The sensitivity of RT4 cells to MMC was decreased after the treatment with rhCXCL5 (P < 0.01), whereas the sensitivity of M-RT4 cells to MMC was increased after CXCL 5 gene knockdown (P < 0.01). As compared with RT4 cells, the expressions of E-cadherin, Cyt-7 and Claudin-1 in M-RT4 cells were significantly decreased (all P < 0.01), but the expressions of N-cadherin, Vimentin, NF-κB1, Rel A, NF- κB2 and Rel B were increased (all P < 0.05). The treatment of rhCXCL up-regulated the expressions of Snail, NF-κB1, Rel A and Rel B in RT4 cells (all P < 0.05). Knockdown of CXCL 5 gene inhibited Snail activation (P < 0.05), up-regulated Cyt-7 expression (P < 0.01), and down-regulated the expressions of NF-κB1, Rel A and Rel B in M-RT4 cells (all P < 0.05). Conclusion: CXCL5 is over-expressed in NMIBC tissues and the chemoresistant bladder cancer cells. Knockdown of CXCL 5 gene expression may reduce the resistance of M-RT4 cells to MMC by reversing EMT, suggesting that CXCL5 is an important factor leading to the intravesical chemoresistance of NMIBC.
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The proliferation,progression and metastasis of pancreatic cancer are complicated processes caused by muhiple factors.C-X-C motif ligand 5 (CXCL5) can recognize and bind to C-X-C motif receptor 2 (CXCR2),and activate or regulate the expression of signaling pathway by autocrine or paracrine pathway in cells.CXCL5/CXCR2 biological axis plays important roles in proliferation,adhesion,progression,metastasis and prognosis of pancreatic cancer,and is also associated with the angiogenesis and lymphangiogenesis of pancreatic cancer.So,direct or indirect regulation of CXCL5/CXCR2 expression in pancreatic cancer can be effective for target therapy of pancreatic cancer.
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Objective: To evaluate the anti-inflammatory property of both glycyrrhizic acid (GA) and glabridin in reducing inflammation to accelerate wound regeneration on 3T3-L1 and NIH-3T3 fibroblast cell lines. Methods: Cell proliferation and viability assay (MTT assay), scratch wound healing assays, and quantitative real-time PCR were conducted to investigate the effects on cell proliferation, cell migration, and expression of CXC chemokine ligand 5 inflammation gene respectively. Results: Results showed that at a low concentration of 1 × 10
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Objective To detect the expressions of chemokine CXCL5 and its receptor CXCR2 in hepatocellular carcinoma (HCC), investigate their relationships between CXCL5, CXCR2 and clinicopathologic features and probe into the significance in the evaluation of prognosis. Methods Immunohistochemical Envision method was utilized to detect the expressions of CXCL5 and CXCR2 in HCC , to explore their relationship between CXCL5, CXCR2 and clinicopathologic features and MVD in HCC. Results (1) The high expression rates of CXCL5 and CXCR2 protein was 64.7% and 68.6%, respectively, significantly higher than those in adjacent tissues of HCC(17.6%, 15.7%, P < 0.05). The overexpression of CXCL5 protein had correlation with tumor size, differentiation, TNM stage and vascular invasion (P < 0.05) and the overexpression of CXCR2 protein did with tumor grade and vascular invasion (P < 0.05). (2) The value of MVD was higher in HCC than that in the adjacent tissues of HCC (P < 0.05), and had positive correlation with the expressions of CXCL5, and CXCR2 proteins. (3) The higher rates of recurrence and metastasis were in the groups of higher expression of CXCL5 and CXCR2 proteins (P < 0.05). Conclusions The overexpressions of CXCL5 and CXCR2 may promote the occurrence and development of HCC as well as the neovasculation in HCC. Therefore, they can be used as markers to evaluate the prognosis of HCC.
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The association between CXCL5 gene polymorphism and obese patients with type 2 diabetes mellitus (T2DM) was explored.The distribution of CXCL5 gene promoter region-156G/C polymorphism revealed no significant difference between normal control group and T2DM group (P＞0.05).The frequency of C allele gene in obesity group was higher than that in non-obesity group(P＜0.05).The results suggest that the CXCL5 promoter gene -156G/C polymorphsim has no relation with T2DM,but it is a risk factor for obesity.
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Objective To investigate the.expression and significance of peritoneal fluid (PF) erythropoietin (EPO) and neutrophils activation peptide-78 (ENA-78)in patients with endometriosis.Methods The levels of EPO and ENA-78 in peritoneal fluid (PF) were measured by means of enzymelinked immunosobent assay (ELISA) in 30 women with endometriosis (9 in Ⅰ ～ Ⅱ stage,21 in Ⅲ～ Ⅳ stage)and 30 without endometriosis.And the relationship between the PF levels of EPO and ENA-78 in patients with endometriosis was analyzed.Results The PF levels of EPO and ENA-78 in women with endometriosis [ (9.272 ± 2.284 ) IU/L,( 2.068 ± 0.794 ) ng/ml ] were significantly higher than those in control group [ (5.759 ±0.502) IU/L,(0.886 ±0.145 ) ng/ml,P ＜0.05].There were no significant differences in the PF levels of EPO between stage Ⅰ ～ Ⅱ [ (9.549 ± 1.996) IU/L] and Ⅲ ～ Ⅳ [ (9.181 ± 1.897) IU/L,P ＞0.05].The ENA-78 concentrations of PF in stage Ⅲ～ Ⅳ [ (2.517 ± 0.518 )ng/ml] were significantly higher than those in stage Ⅰ ～ Ⅱ[ ( 1.137 ±0.169)ng/ml,P ＜0.05].There were no obvious correlations in PF EPO and ENA-78 in patients with endometriosis.Conclusions The excessive expression of EPO and ENA-78 in the PF of women with endometriosis suggests that EPO and ENA-78 may be an important role in the pathogenesis of endometriosis.
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Objective To investigate the level of chemotactic factors(CXCL5 and CXCL8)in hepatic fibrosis and cirrhosis tissue.Methods Hepatic tissues were obtained from 9 patients with hepatic hemangioma (hepatic hemangioma group),10 patients with liver fibrosis(liver fibrosis group)and 11 patients with liver cirrhosis(1iver cirrhosis group)at Nanfang Hospital from May 2008 to May 2009.The contents of CXCL5 and CXCL8 in hepatic tissue were assayed by ELISA.All data were analyzed by one-way ANOVA,Pearson rank correlation or Spearman correlation.Results The contents of CXCL5 and CXCL8 were(0.8±0.7)ng/g and(6.2±3.7)ng/g in hepatic hemangioma group,(2.0±2.0)ng/g and(11.6±3.5)ng/g in liver fibrosis group and (17.1±4.8)ng/g and(12.3±3.9)ng/g in liver cirrhosis group,with significant difference among the 3 groups (F=60.050,7.690,P＜0.05).The expression of CXCL5 was correlated with the content of alanine aminotransferase(ALT),aspartate aminotransferase(AST)and prothrombin time(PT)(r=0.502,0.468,0.523,P＜0.05):the expression of CXCL8 was correlated with the content of ALT,AST.total bilirubin and PT(r=0.477,0.504,0.537,0.431,P＜0.05).Conclusions With the aggravation of hepatic fibrosis,the contents of CXCL5 and CXCL8 are increased with different patterns.The changes of CXCL5 and CXCL8 are related with the injury of liver,but the changes of CXCL5 and CXCL8 do not correspond with the degree of the injury of liver.