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Objective:To explore the effect of esophageal squamous cell carcinoma-related long non-coding RNA (lncRNA) ESCCAL-1 on the polarization of THP-1 cells-derived macrophages.Methods:The esophageal cancer cell line KYSE450 was divided into 5 groups: KYSE450 group (normal KYSE450 cells), shRNA-ESCCAL-1 group (infected with knockout ESCCAL-1 lentivirus), shRNA-NC group (infected with interference control lentivirus), OE-ESCCAL-1 group (infected with overexpressing ESCCAL-1 lentivirus) and OE-NC group (infected with overexpressed control lentivirus). The expression of ESCCAL-1 was detected by real-time quantitative polymerase chain reaction (qRT-PCR). After co-culture of cells in each group with THP-1 cells-derived macrophages, flow cytometry was used to detect the expressions of THP-1 cells-derived macrophages M1 polarization markers HLA-DR, iNOS, CD86 and M2 polarization markers Arg-1, CD163, CD206, and inflammatory cytokines.Results:After THP-1 cells were stimulated with 100 ng/ml phorbol ester for 48 hours, the cells grew adherently, and the expression levels of CD11b and CD36 increased, indicating that THP-1 cells were successfully differentiated into macrophages. After THP-1 cells-derived macrophages were co-cultured with esophageal cancer KYSE450 cell line treated differently for 24 hours, there were no significant differences in the expressions of M1 polarization markers HLA-DR, iNOS and CD86 between shRNA-ESCCAL-1 group and shRNA-NC group or between OE-ESCCAL-1 group and OE-NC group (all P > 0.05). Compared with shRNA-NC group, the expressions of M2 polarization markers Arg-1, CD163 and CD206 in shRNA-ESCCAL-1 group decreased [8.54±0.29 vs. 11.83±0.69, 12.0±0.3 vs. 24.5±0.8, 2.05±0.23 vs. 14.54±1.10], and the differences were statistically significant ( t values were 7.636, 27.38 and 19.31, all P < 0.01); compared with the OE-NC group, the expressions of M2 polarization marker Arg-1, CD163 and CD206 in OE-ESCCAL-1 group increased [32.60±1.14 vs. 14.20±0.20, 43.7±1.5 vs. 25.1±1.2, 35.8±0.7 vs. 13.6±0.6], and the differences were statistically significant ( t values were -27.58, -17.24 and -43.98, all P < 0.01). Compared with shRNA-NC group, the expression level of interferon-γ in shRNA-ESCCAL-1 group decreased [(6.3±1.5) pg/ml vs. (20.0±2.6) pg/ml, t = 7.75, P = 0.001]; compared with OE-NC group, the expression level of interleukin-1RA in OE-ESCCAL-1 group increased [(3 167±306) pg/ml vs. (467±176) pg/ml, t = -13.27, P < 0.01]. Conclusions:Esophageal squamous cell carcinoma-related lncRNA ESCCAL-1 can promote the M2 polarization of macrophages.
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Objective@#To investigate the relationship between UGT1A1*6, UGT1A1*28, UGT1A1*60 and UGT1A1*93 polymorphisms and irinotecan-induced severe adverse reactions(grade 3-4 delayed diarrhea and neutropenia) in Chinese cancer patients.@*Methods@#A total of 141 cancer patients treated with irinotecan were enrolled in this study. Peripheral venous blood was collected and genomic DNA was extracted. The genetic polymorphisms of UGT1A1*6, UGT1A1*28, UGT1A1*60 and UGT1A1*93 were analyzed by PCR and direct sequencing. The adverse reactions during chemotherapy were observed and recorded. The incidence of severe adverse reactions was compared among patients with different genotypes.@*Results@#Among 141 patients, the cases with UGT1A1*6 GG, GA and AA genotypes were 71, 54 and 16, while those with UGT1A1*28 TA6/6, TA6/7 and TA7/7 genotypes were 105, 33 and 3, respectively. The cases with UGT1A1*60 AA, AC and CC genotypes were 52, 80 and 9, while those with UGT1A1*93 GG, GA and AA genotypes were 105, 32 and 4, respectively. The patients with grade 3-4 delayed diarrhea and neutropenia were 23 and 56, respectively. Multivariate logistic regression analysis showed that UGT1A1*6 and UGT1A1*60 genetic polymorphisms were independent factors influencing the occurrence of grade 3-4 delayed diarrhea. The risk of grade 3-4 delayed diarrhea in homozygous AA carriers of UGT1A1*6 increased 3.79 times compared with that in wild-type GG carriers (95%CI: 1.35-10.67). Moreover, the risk of grade 3-4 delayed diarrhea in homozygous CC carriers of UGT1A1*60 was 20.42 times compared with that in wild-type AA carriers (95%CI: 3.52-118.33). In addition, UGT1A1*28 genetic polymorphism was an independent factor of the occurrence of grade 3-4 neutropenia. The patients with homozygous TA7/7 carriers of UGT1A1*28 had an 1.61 times higher risk of grade 3-4 neutropenia compared with those with wild-type TA6/6 carriers (95%CI: 1.44-12.65). There was no correlation between UGT1A1*93 genetic polymorphism and severe adverse reactions caused by irinotecan.@*Conclusion@#The cancer patients who carried UGT1A1*6, UGT1A1*28 and UGT1A1*60 gene polymorphisms have high risk of severe adverse events caused by irinotecan-based chemotherapy.
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Aim TostudytheeffectsofCuO,ZnOand TiO2 nanoparticles on the viability and metastatic po-tential of EC-9706 and EC-109 esophageal squamous carcinomacelllineinvitro.Methods Characteristics of CuO,ZnO and TiO2 nanoparticles were detected u-sing transmission electron microscope (TEM)and dy-namic light scattering (DLS ).EC-9706 and EC-109 cells were treated with different concentrations of CuO, ZnO and TiO2 (5 ~80 mg · L-1 ).The cell prolifera-tion was analyzed by MTT assay.The cell cycle and apoptotic rates were determined by flow cytometry (FCM).The cell invasion was assayed in Transwell chambers.The expression of Bcl-2 and caspase-3 pro-tein in cells was detected by Western blot method.Re-sults CuO,ZnOandTiO2nanoparticleswerespheri-cal with primary particle size 12,20. 6,12 nm.The particles were agglomerated in water and cell culture medium with negative charge.CuO and ZnO nanoparti-cles induced decreases in EC-9706 and EC-109 cell vi-ability dose-dependently.After exposed to increasing concentrations of CuO and ZnO nanoparticles,the cell cycle analysis revealed a decreasing proportion of cells in G2/Mand S phase,and up-regulation of the cells in G0/G1 phase.Apoptotic cells also increased along with decreased cell invasion upon CuO and ZnO treatment. Nanoparticles treatment after 48 h, the activated caspase-3 expression quantity increased significantly and the Bcl-2 expression quantity decreased obviously (P<0. 05 )compared with control group.TiO2 nanop-articles had no obvious effect on the EC-9706 and EC-109 cell proliferation,cell cycle,apoptosis and inva-sion.Conclusion ComparedwithTiO2,CuOand ZnO nanoparticles can inhibit EC-9706 and EC-109 cell viability and metastatic potential,the mechanism of action involves cell cycle arrest in G0/G1 phase and apoptosis.These findings can help the development of nanoparticles as anti-cancer therapeutics for esophageal cancer.