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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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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 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.