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Ge X.,Nanjing Medical University | Zhen F.,Nanjing Medical University | Yang B.,Nantong Tumour Hospital | Yang X.,Nanjing Medical University | And 7 more authors.
Journal of International Medical Research | Year: 2014

Objective: To determine if the pretreatment of hypoxic human oesophageal carcinoma cell lines (EC109, TE1 and KYSE170) with ginsenoside Rg3 (Rg3) increases their radiosensitivity to X-rays. Methods: The growth inhibitory effect of different Rg3 concentrations was measured using the 3-(4,5-dimethylthiazol-2-yl)-2,5- diphenyltetrazolium bromide (MTT) assay. Radiation sensitivity was measured using a clone formation assay and flow cytometry was used to measure the effects of Rg3 on radiation-induced apoptosis.Western blot analysis was used to measure the effects of Rg3 on the levels of hypoxia inducible factor (HIF)-1α and vascular endothelial growth factor (VEGF). Results: Rg3 inhibited EC109, TE1 and KYSE170 cell growth in a dose- and time-dependent manner. Pretreatment with 10 mmol/ml Rg3 increased EC109, TE1 and KYSE170 radiosensitivity. Rg3 plus radiation significantly increased the apoptosis rate compared with radiation alone. Rg3 also decreased VEGF and HIF-1α protein levels in EC109 cells in a dose-dependent manner. The combination of Rg3 and radiation increased the fragmentation of double-stranded DNA. Conclusion: Rg3 enhanced the radiosensitivity of human oesophageal carcinoma cell lines cultured under hypoxic conditions possibly by downregulating VEGF and HIF-1α protein levels.© The Author(s) 2014 Reprints and permissions: sagepub.co.uk/journalsPermissions.nav.


Wu D.,Nantong University | Zhang X.,Nantong University | Zhao M.,Nantong Tumour Hospital | Zhou A.-L.,Nantong University
Sheng li xue bao : [Acta physiologica Sinica] | Year: 2015

The present study aimed to investigate the role of the Toll-like receptor 4 (TLR4)/nuclear factor κB (NF-κB) signaling pathway in the accumulation of amyloid β protein (Aβ) in primary hippocampal neurons of rats. The purity of these cultured neurons was determined by using immunofluorescence techniques. Lipopolysaccharide (LPS, a TLR4 ligand) or CLI-095 (a TLR4 inhibitor) was used to activate or inhibit TLR4 signaling, respectively. Pyrrolidine dithiocarbamate (PDTC), on the other hand, was used to inhibit NF-κB, a downstream effector of the TLR4 signaling pathway. The contents of tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β) and Aβ1-42 in the supernatant were assessed by enzyme-linked immunosorbent assay (ELISA). The mRNA levels of TNF-α, IL-1β, a disintegrin and metalloproteinase domain-containing protein 10 (ADAM10), β-site APP cleaving enzyme 1 (BACE-1), Presenilin-1 (PS-1), and β-amyloid precursor protein (β-APP) were examined by real-time quantitative PCR (RT-qPCR). The protein levels of ADAM10, BACE-1, PS-1 and β-APP were examined by Western blotting. Meanwhile, the levels of TLR4 mRNA and protein in hippocampal neurons were tested by RT-qPCR and Western blotting, respectively, after stimulation with Aβ1-42 at different concentrations. We observed that the purity of cultured hippocampal neurons after being cultured for 7 days was above 95%. Compared with untreated neurons, LPS-treated neurons showed higher expression levels of TNF-α, IL-1β, BACE-1, PS-1, β-APP, and Aβ1-42, but a lower expression level of ADAM10. These effects were reversed upon pre-treatment with CLI-095 or PDTC. Furthermore, TLR4 expression was upregulated in the presence of Aβ1-42. Taken together, these results provide evidence that elevation in the level of inflammatory cytokines accompanies the activation of TLR4 signaling, and that the consequent downregulation of ADAM10 and upregulation of BACE-1/PS-1 are likely responsible for the accumulation of β-APP and Aβ, which in turn increases TLR4 level to create a positive feedback loop that may constitute the basis for the progression of Alzheimer's disease.


Han L.,Nantong Tumour Hospital | Liu W.,Nantong Tumour Hospital | Chen Y.,Nantong Tumour Hospital | Wu H.,Nantong University | And 2 more authors.
Neoplasma | Year: 2015

The growth-regulated oncogene α (GROα) , which is also designated as CXC chemokine ligand 1 (CXCL1), was first identified as an autocrine growth factor in human malignant melanoma. It is involved in tumor development and invasion, and is highly expressed in various human cancers. However, little is known about the association between GROα expression and the clinical attributes of laryngeal squamous cell carcinoma (LSCC). One-step quantitative reverse transcription-polymerase chain reaction (qRT-PCR) and immunohistochemical staining of tissue microarrays were employed to evaluate the relationship between GROα expression and LSCC clinicopathological attributes. GROα mRNA and protein expression levels were significantly greater in LSCC than in non-cancerous tumor-adjacent tissues. GROα protein expression in LSCC was also significantly associated with TNM stage, lymph node metastasis, and histopathological grade. Kaplan-Meier and Cox multi-factor analyses suggested that increased GROα expression and positive lymph node metastasis were significantly associated with the poor survival of LSCC patients. These data indicate that GROα may be a novel prognostic marker of LSCC. © 2015 Cancer Research Institute Slovak Acad. of Sciences. All rights reserved.


PubMed | Nantong Tumour Hospital and Nantong University
Type: Journal Article | Journal: Sheng li xue bao : [Acta physiologica Sinica] | Year: 2015

The present study aimed to investigate the role of the Toll-like receptor 4 (TLR4)/nuclear factor B (NF-B) signaling pathway in the accumulation of amyloid protein (A) in primary hippocampal neurons of rats. The purity of these cultured neurons was determined by using immunofluorescence techniques. Lipopolysaccharide (LPS, a TLR4 ligand) or CLI-095 (a TLR4 inhibitor) was used to activate or inhibit TLR4 signaling, respectively. Pyrrolidine dithiocarbamate (PDTC), on the other hand, was used to inhibit NF-B, a downstream effector of the TLR4 signaling pathway. The contents of tumor necrosis factor- (TNF-), interleukin-1 (IL-1) and A1-42 in the supernatant were assessed by enzyme-linked immunosorbent assay (ELISA). The mRNA levels of TNF-, IL-1, a disintegrin and metalloproteinase domain-containing protein 10 (ADAM10), -site APP cleaving enzyme 1 (BACE-1), Presenilin-1 (PS-1), and -amyloid precursor protein (-APP) were examined by real-time quantitative PCR (RT-qPCR). The protein levels of ADAM10, BACE-1, PS-1 and -APP were examined by Western blotting. Meanwhile, the levels of TLR4 mRNA and protein in hippocampal neurons were tested by RT-qPCR and Western blotting, respectively, after stimulation with A1-42 at different concentrations. We observed that the purity of cultured hippocampal neurons after being cultured for 7 days was above 95%. Compared with untreated neurons, LPS-treated neurons showed higher expression levels of TNF-, IL-1, BACE-1, PS-1, -APP, and A1-42, but a lower expression level of ADAM10. These effects were reversed upon pre-treatment with CLI-095 or PDTC. Furthermore, TLR4 expression was upregulated in the presence of A1-42. Taken together, these results provide evidence that elevation in the level of inflammatory cytokines accompanies the activation of TLR4 signaling, and that the consequent downregulation of ADAM10 and upregulation of BACE-1/PS-1 are likely responsible for the accumulation of -APP and A, which in turn increases TLR4 level to create a positive feedback loop that may constitute the basis for the progression of Alzheimers disease.

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