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Objective:To explore the effect and mechanism of fentanyl on promoting the stemness of breast cancer cell.Methods:From January 2018 to October 2019, human breast cancer cell line BT549 was used as the in vitro research object. Breast cancer cell BT549 was pretreated with 0.01 and 0.10 μmol/L fentanyl. Sphere formation assay and colony formation assay were performed to investigate the role of fentanyl on breast cancer cell stemness. Fluorescent quantitative polymerase chain reaction (FQ-PCR) was used to detect the mRNA level of stemness-related transcription factors gender-determining area Y box protein 2 (Sox2), octamer binding transcription factor 4 (Oct4) and Nanog. Western blotting assay was performed to determine the level of Wnt3a/β-catenin pathway-related markers Wnt3a, phosphorylated glycogen synthase kinase-3β (p-GSK-3β), glycogenase kinase-3β (GSK-3β) and β-catenin. After down-regulating Wnt3a, western blotting assay and sphere formation assay were performed.Results:The sphere diameter, colony formation rate and the expression of Sox2 mRNA, Oct4 mRNA, Nanog mRNA, Wnt3a, p-GSK-3β, GSK-3β and β-catenin in 0.01 and 0.10 μmol/L fentanyl-treated breast cancer cell were significant higher than those in blank control: (131.22 ± 1.06) and (636.37 ± 0.02) μm vs. (72.68 ± 0.13) μm, (41.33 ± 0.03)% and (60.58 ± 1.08)% vs. (20.93 ± 0.15)%, 2.25 ± 0.20 and 3.82 ± 0.84 vs. 1.00, 1.87 ± 1.06 and 3.35 ± 0.04 vs. 1.00, 2.85 ± 0.03 and 4.36 ± 0.50 vs. 1.00, 1.82 ± 0.03 and 2.57 ± 0.42 vs. 1.00, 2.04 ± 0.13 and 2.81 ± 0.05 vs. 1.00, 1.62 ± 0.17 and 2.93 ± 0.06 vs. 1.00, 2.15 ± 0.02 and 3.54 ± 0.21 vs. 1.00, the indexes in 0.10 μmol/L fentanyl-treated breast cancer cell were significantly higher than those in 0.01 μmol/L fentanyl-treated breast cancer cell, and there were statistical differences ( P<0.01). After down-regulating Wnt3a, the expressions of p-GSK-3β, GSK-3β, β-catenin, Sox2, Oct4 and sphere diameter were significantly lower than those in blank control: 0.12 ± 0.05 vs. 1.00, 0.53 ± 0.06 vs. 1.00 and 0.24 ± 0.21 vs. 1.00, 0.28 ± 0.10 vs. 1.00 and 0.06 ± 0.01 vs. 1.00, (18.14 ± 0.30) μm vs. (74.32 ± 0.12) μm, and there were statistical differences ( P<0.01). Conclusions:Fentanyl promotes breast cancer cell stemness by Wnt3a/β-catenin signaling pathway.
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Objective To explored the effects of fentanyl on cell proliferation of H1299 cells, Methods After treating H1299 cells with different concentrations of fentanyl (0.001, 0.010, 0.100, 1.000 μM) for 12, 24, 48, 72 h, cell viability was detected by CCK-8 assay; the rate of cell apoptosis was determined by DAPI staining; the expression levels of Bax, Bcl-2, p-AKT and AKT protein were measured by Western blotting; Caspase-3 activity was determined by Caspase-3 activity assay kit. Results Compared with the control group, fentanyl obviously inhibited the viability of H1299 cells in a dose and time dependent way. Moreover, treatment with different concentrations of fentanyl(0.001, 0.010, 0.100, 1.000 μM) for 12, 24, 48, 72 h, the apoptosis rate of H1299 cells were significantly increased, The level of Bcl-2 protein reduced the level of Bax protein, and the activity of Caspase-3 in H1299 cells were increased after treatment with fentanyl (0.010, 0.100, 1.000 μM) for 48 h, Furthermore, fentanyl markedly inhibited p-AKT/AKT activity of H1299 cells. Conclusions Fentanyl can inhibit cell proliferation and promote cell apoptosis of human lung cancer, and its mechanism may be related to inhibition of AKT activation ,
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Objective To investigate the effect of intrathecal parecoxib sodium in bone tumor pain of rats. Methods Bone tumor pain was induced by injection of MRMT-1 tumor cells (1 × 104/L) into the tibia of female SD rats under sevoflurane anesthesia. The development of bone tumor was monitored by radiological study, and histological sections stained with hematoxylin and eosin. At 3 d after MRMT-1 tumor cell injection, a PE-10 catheter was inserted into the intrathecal space for drug administration. At 10 d after MRMT-1 tumor cell injection, rats were randomly divided into 5 equal groups, control group, parecoxib sodium 0.1, 0.3, 1.0 and 3.0 g/L group. For pain assessment, a withdrawal threshold was measured using von Frey filament being applied to the tumor cell inoculation site. The effects of intrathecal saline or parecoxib sodium were investigated. Results Intra-tibial injection of MRMT-1 tumor cells produced a bone tumor in radiologic and pathologic findings.Also, the paw withdrawal threshold was significantly decreased(mechanical allodynia).Percentage of the maximal possible effect (% MPE) of control group and parecoxib sodium 0.1, 0.3, 1.0 and 3.0 g/L group was (13.89 ± 4.17)%,(7.54 ± 3.91)%,(57.47 ± 11.47)%,(85.72 ± 9.42)% and(100.00 ± 0.00)%,compared with control group, intrathecal parecoxib sodium dose-dependently increased the withdrawal threshold (P<0.05).Conclusions Intrathecal parecoxib sodium reduces bone tumor-related pain behavior.
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Objective To evaluate the occurrence and management of complications following mini- mally invasive percutaneous nephrostolithotomy (MPCNL).Methods The data of 436 cases of MPCNL from December 2001 to March 2005 were reviewed,including 249 male cases and 187 female cases.Their age ranged from 14 to 71 years with a mean of 41.6 years.Of the 436 cases,314 cases had simple nephrolith- iasis,79 had nephrolithiasis combined with upper ureterolithiasis,27 had unilateral upper ureterolithiasis,and 16 had bilateral upper ureterolithiasis.Results Among the 436 cases of MPCNL,complications occurred in 27 cases (6.2%).Of the 27 cases,5 had massive hemorrhage,which was cured by closing meatus of nephric fistula,cleaning out the blood clot within the bladder with instrument and transfusing blood;2 cases had pleural effusion,11 cases had hydroperitoneum,8 eases had renal perforating injury,and they were cured by puncture and drainage;1 case had pyocalix,which was cured by using effective antibiotics,re-punc- turing kidney and postponing pulling out the fistula catheter.Conclusions MPCNL is a minimally inva- sire operation.However,it has obvious risk if the surgeon's skill is not proficient.The improvement in the pre- vention and management of complications can promote the application of this procedure.