ABSTRACT
Objective To evaluate the effects of remifentanil on the metastasis of human lung adenocarcinoma cells and expression of interleukin-7 receptor (IL-7R).Methods Human adenocarcinoma cell line A549 cells were seeded in 24-well plates at a density of 2× 105 cells/ml (72 wells).The cells were divided into 4 groups (n =18 each) using a random number table:normal control group (group C) and remifentanil 2,4 and 6 ng/ml groups (group R2,group R4,group R6).In R2,R4 and R6 groups,remifentanil at the final concentrations of 2,4 and 6 ng/ml was added,respectively,and the cells were incubated for 4 h.The equal volume of normal saline was given instead of remifentanil in group C.The cells were collected at 24 h after the following incubation.The invasion of cells was determined by Transwell invasion assay,and the invaded cells were counted.The migration of cells was determined by cell scratch test,and cell migration rates were calculated.The expression of IL-7R in cells was detected by Western blot.Results Compared with group C,the number of invaded cells and cell migration rates were significantly decreased in R2,R4 and R6 groups,and the expression of IL-7R was down-regulated in R4 and R6 groups (P＜0.05).Compared with group R2,the number of invaded cells and cell migration rates were significantly decreased,and the expression of IL-7R was down-regulated in R4 and R6 groups (P＜0.05).Compared with group R4,the number of invaded cells and cell migration rates were significantly decreased,and the expression of IL-7R was down-regulated in group R6 (P＜0.05).Conclusion Remifentanil can inhibit metastasis of human lung adenocarcinoma cells and down-regulate the expression of IL-7R.
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ObjectiveTo investigate whether CD4+ CD25+ CD127dim/- regulatory T lymphocytes (Treg) can induce the proliferation of hepatic stellate cells (HSC) and expression of fibrosis-related factors on HSC in vitro and further to explore the mechanism of Treg inducing fibrogenesis. MethodsHSC LX-2 cells were subcultured.CD4+ CD25+ CD127dim/- cells were purified using magnetic cell separation. The HSC were co-cultured with Treg by direct contact or by Transwell system in vitro. The HSC cultured alone was used as control. Cell proliferation was measured by CCK-8 assay.The expression of transforming growth factor (TGF)-β1 was detected by enzyme-linked inmunosorbent assay (ELISA), and the expressions of hyaluronic acid (HA) and precollagen Ⅲ (PC Ⅲ ) were detected by radio immunoassay (RIA). The data were analyzed by LSD-t test. ResultsHSC proliferation was strongest when Treg∶ HSC= 1.5∶ 1. The absorbance in direct contact co-culture group and Transwell system co-culture group was (0. 713±0. 032) cpm and (0. 735±0. 028) cpm, respectively, both of which were higher than that in control group [(0. 677 ± 0. 029) cpm](t = 5. 4003 and 8. 7878,respectively; both P＜0. 01). The concentrations of TGF-β1 in the supernatant were (781. 59 ±76.45) pg/mL and (813. 53±60. 62) pg/mL, respectively in direct contact co-culture group and Transwell system co-culture group, which were significantly higher than that in control group [(722.51±59. 66) pg/mL](t = 4.0014 and 6. 1653, respectively; both P＜0.01).The concentrations of HA were (433. 575±27.90) ng/mL and (445.40±23.73) ng/mL, respectively in direct contact co-culture group and Transwell system co-culture group, which were higher compared to that in control group [-(415. 83±19.44) ng/mL](t =3. 3124 and 5. 5231, respectively; both P＜0.01). Likewise, the concentrations of PCⅢ were (21. 93± 1.71) and (23. 125± 1.87) ng/mL in direct contact group and Transwell group, respectively compared to (20. 10± 1.49) ng/mL in control group (t = 4. 8082 and 7. 9436, respectively; both P ＜ 0.01).Furthermore, the absorbance,concentrations of TGF-β1, HA and PC Ⅲ in Transwell co-culture group were all higher compared to direct contact group (t = 3. 3875, 2.1639, 2. 2107 and 3.1354, respectively; all P＜0. 05).ConclusionsThe cell proliferation and the expressions of fibrosis-related factors in HSC increase greatly after co-cultured with CD4+ CD25+ CD127dim/-Treg. Therefore, Treg may play an important role in inducing liver fibrogenesis.
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ObjectiveTo study the relationship between the susceptibility of mutiple sclerosis (MS) and single nucleotide polymorphisms (SNP) rs6897932 of interleukin-7 receptor alpha (IL-7RA) in Asian.MethodsThe SNP rs6897932 in the IL-7RA gene was genotyped by real time polymerase chain reaction (PCR) using TaqMan SNP Genotyping Assays. 78 cases with MS and neuromyelitis optica (NMO), 187 patients with non-NMO MS and 158 healthy controls were enrolled.ResultsThe frequencies of both the C allele and the CC genotype of SNP rs6897932 in the IL-7RA gene in non-NMO MS patients were higher than those of healthy controls(89.3％ vs. 79.8％, OR=2.12, 95％CI:1.38-3.25 P＜0.01; 78.6％ vs. 63.3％, OR=2.13,95％CI:1.32-3.43, P＜0.01).However, there was no significant difference in the frequency of either the C allele or the CC genotype between control group and NMO patients.ConclusionsIL-7RA gene is one of susceptibility gene for MS in Asian. C allele was presumed as a risk factor of MS while T allele might be a protective factor.
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Objective The aim of the study WSfl to establish the method using CDl27 as the new biomarker to identify regulatory T cells(Treg cells)and apply the CD 127 to detect the Treg cells in patients with gastric cancer.Methods The phenotypes of Treg cells were analyzed using five-color flow cytometry method Foxp3.FITC/CD127-PE/CD4-PerCP/CD25-APC/CD3-PC7.The mRNA and protein expression of Foxp3 in isolated CD+4 CD25high CD127-/low Treg cells were detected.The relationships between Foxp3 and CD127 protein expression in CD4+ T cells from aduh human peripheral blood were investigated.PBMCs,Ascltes,turnor-infihration lymphocyte and tumor-draining lymph nodes in 35 patients with gastric cancer and PBMCs in 20 normal healthy donors were evaluated for the proportion of Treg ceils,as well as the percentage ot the total CD +4 cells.Results CD4+ CD25 high CD-127 low ceils expressed the high Foxp3 in protein level(87.1%)and mRNA level.Within the CD4+CD+25 population,there was a significant correlation between Foxp3 and the CD127low phenotype(r=0.985,P<0.01).Compared with healthy olunteers,patients with gastric malignancies had a higher proportion of CD4+4 Cdhigh 25 CD-low127 cells in peripheral blood(t=2.542,P<0.05).The Dereentages of Treg cells were more abundant in ascites(t=2.357,P<0.05),TIL(t=6.174,P<0.01) and tumor-draining lymph nodes(t=5.481,P<0.01)of individuals with gastric cancer than that in their blood.There were significant differences in the prevalence of Treg ceils between the early and advanced disease stages in gastric cancer[(6.04±2.31)%in stageⅠ+Ⅱ VB(10.16±2.29)% Ⅲ+Ⅳ,t=2.473,P<0.05].Conclusions The CD127 biomarker can be used to selectively enrich human Treg cells for in vitro functional studies.The populations of CD4+ CD high25 CD127-/low Treg cells increased ith tumor stage in individuals with gastric cancer.