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Objective To examine the expressions of IKKε protein in the specimens and cells of epithelial ovarian cancer and investigate the effect of IKKε inhibitor on cell proliferation and apoptosis. Methods (1) A total of 118 cases of patients with the median age of 59 who have accepted surgical treatment due to ovarian cancer in the First Affiliated Hospital of Dalian Medical University from January 2006 to April 2013 were selected. Twenty cases of patients with the median age of 55 who have accepted hysterectomy and salpingo-oophorectomy due to uterine leiomyoma during the same period were selected as the control. The expressions of IKKε protein were detected by immunohistochemistry in normal ovarian tissues and epithelial ovarian cancer specimens,and the relationship between the expressions of IKKε and the clinical features of patients was analyzed. IKKε protein was determined by western blot in various ovarian cancer cells, including SKOV3, OV2008, C13, A2780S, A2780CP, OV4, OV5, OV8, and CAOV3 treated with or without IKKε inhibitor. The cellular proliferation and apoptosis of ovarian cancer cells after 48 hours treatment of IKKε inhibitor were analyzed by methyl thiazolyl tetrazolium (MTT) assay and flow cytometry, respectively. Results (1) The immunohistochemical results showed that IKKε was highly expressed in epithelial ovarian cancer specimens with the expression rate 66.1% (78/118), compared with normal ovarian tissue with the expression rate 35.0% (7/20), which exhibited statistically significant difference (χ2=6.993, P=0.008). The expression of IKKε protein was correlated with International Federation of Gynecology and Obstetrics (FIGO) stage, histological grade, the level of CA125 in preoperative serum and distribution of the tumor (P0.05). (2) IKKε was widely overexpressed in different levels in SKOV3, OV2008, C13, A2780S, A2780CP, OV4, OV5, OV8, and CAOV3 cells, and the expression of IKKε decreased as the increase of the concentration of IKKε inhibitor (0.1 and 0.5 μmol/L) in OV2008, C13, A2780S, and A2780CP cells after 48 hours treatment. Different concentrations of IKKε inhibitor (0.05, 0.1, 0.5, 1, 5, 10, and 25 μmol/L) significantly inhibited the proliferation of OV2008, C13, A2780S, A2780CP, and SKOV3 cells in a concentration-dependent manner (P<0.05), and the half maximal inhibitory concentration (IC50) was 0.43, 0.86, 0.10, 0.19, and 0.24 μmol/L, respectively. The cell apoptotic rate of OV2008, C13, A2780S, A2780CP, and SKOV3 cells was significantly increased after 48 hours treatment of IKKεinhibitor with the concentration of 0.1 and 0.5 μmol/L (P<0.05). Conclusions The IKKε protein in epithelial ovarian cancer specimens and cells is overexpressed. IKKε inhibitor could inhibit cellular proliferation and induce apoptosis in a concentration-dependent manner. Together, the result indicated that IKKε may be a candidate target for the treatment of ovarian cancer in future.
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Objective To investigate the effecof Iκkinases(IKK) inhibitoIKK16 on the proliferation and apoptosiof hu-man glioblastomcell line U87 and itpossible mechanism .MethodHuman glioblastomcell line U87 wacultured in vitro and divided into 4 group:blank group(withouintervention treatmen) ,control group(1% dimethyl sulfoxide) ,low dose IKK group (70 nmol/L IKK16) and high dose IKK group(200 nmol/L IKK16) .The Mtassay waused to detecthe effecof IKK16 on the proliferation of U87 cellathe low concentration and the high concentration .Western blowaused to investigate the effecof IKK16 on the expression of p65 ,cyclinD1 ,caspase-3 and bcl-2 .ResultThe U87 proliferation rate had no statistical difference be-tween the blank group and the control group(P＞0 .05) .Compared with the control group ,IKK16 could significantly suppresthe proliferation of U87 cell(P＜0 .05) .Athe same time ,IKK16 significantly reduced the expression of p65 ,CyclinD1 and Bcl-2 ,and increased the expression of caspase-3(P＜0 .05) with dose-dependenmanne.Conclusion IKK16 could suppresthe proliferation of human glioblastomcell line U87 and promote the apoptosi.
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Objective To explore the effect of I kappa B kinase (IKK) on liver during hepatic ischemia reperfusion (HIR) in rats.Methods Wister rats were divided randomly into HIR group in which hepatic reperfusion was given after 60 minutes of ischemia by interruption of the arterial and portal venous blood supply to the left lobes and middle lobes of the liver; HIR + PDTC treatment group in which PDTC (120 mg/kg) were injected via the dorsum vein of penis before ischemia reperfusion; and sham control group in which midline laparotomy was performed without vascular occlusion and treatment.Expression levels of IKK were measured with In situ hybridization(ISH).The NF-κB activities were determined with EMSA.Expression levels of TNF-α were measured with immunohistochemistry (IH).Serum levels of ALT were measured.Results Expression level of IKK was increased markedly from 0 to 12h and peaked 6h after reperfusion in HIR group.NF-κB was activated 0 ～ 12h after reperfusion and activities of NF-κB were maximal 6h after reperfusion in HIR group rats compared with sham control group.Expression level of TNFα was increased markedly from 0 to 12h and peaked 6h after reperfusion in HIR group.Serum levels of ALT were increased significantly after reperfusion in H1R group.Expression level of IKK was lowered markedly in HIR + PDTC group from 0 to 12h after reperfusion.NF-κB activities were significantly lower in HIR +PDTC group than in HIR group from 0 to 12h after reperfusion.Expression level of TNF-α was lowered markedly in HIR + PDTC group as compared with HIR group from 0 to 12h after reperfusion.Serum level of ALT was decreased significantly after reperfusion in HIR + PDTC group as compared with HIR group.Conclusion HIR can activate IKK-β which promotes the activation of NF-κB,then NF-κB results in upregulation transcription of TNF-α gene which gives rise to the release of other inflammatory cytokines and triggers uncontrolled inflammatory response,and induces hepatic injury.Blocking IKK-NF-κB pathway may be an effective approach to checking the generation and development of ALI,PDTC plays important prophylaxis and treatment roles in hepatic injury after HIR.
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Objective To observe the effects of gypenosides (GP) on nuclear factor κB (NF-κB) and p38 mitogen activated protein kinase (p38MAPK) signaling pathways in photodamaged skin of mice,and to explore the mechanisms underlying the protective effects of GP against photodamage.Methods Eighty Balb/C female mice were randomly divided into 8 groups: blank control group receiving no treatment,ultraviolet B (UVB) model group receiving UVB irradiation for 60 seconds,GP group Ⅰ receiving topical GP treatment followed by UVB irradiation,GP group Ⅱ receiving UVB irradiation followed by topical GP treatment,VitE group Ⅰ receiving topical VitE treatment followed by UVB irradiation,VitE group Ⅱ receiving UVB irradiation followed by topical VitE treatment,matrix group Ⅰ receiving topical matrix treatment followed by UVB irradiation,matrix group Ⅱ receiving UVB irradiation followed by topical matrix treatment.UVB irradiation lasted 60 seconds at one time,and was given once every other day for 7 times to establish a skin model of photodamage.The interval between irradiation and topical treatment was 30 minutes in all the groups except the control group and UVB model group.After the last treatment,mice were sacrificed.Western blot was performed to measure the protein expressions of inhibitor of NF-κB(IκB),inhibitor of NF-κB kinase (IKK),p38MAPK as well as phosphorylated p38MAPK (pp38) in skin tissue from the mice.Results No expressions of IκB or IKK were observed in the blank control group.The expression level of IκB was 0.40 ± 0.07 in UVB model group,significantly lower than that in GP group Ⅰ (1.63 ± 0.85,P ＜ 0.05) and GP group Ⅱ (0.90 ± 0.40,P ＜ 0.05),whereas the level of IKK protein was higher in UVB model group than in the GP group Ⅰ and Ⅱ (2.01 ± 1.75vs.0.23 ± 0.12 and 0.45 ± 0.29,both P ＜ 0.05).No significant difference was observed in the expression of IκB or IKK proteins between the GP group Ⅰ,Ⅱ,VitE group Ⅰ and Ⅱ or in the expression of p38MAPK between the 8 groups.The phosphorylated p38MAPK expression in UVB model group was significantly higher than that in GP group Ⅰ and Ⅱ (0.835 ± 0.049 vs.0.425 ± 0.054 and 0.571 ± 0.090,both P＜ 0.05),but similar to that in VitE groups.Conclusions UVB can activate NF-κB and phosphorylated p38MAPK signaling pathways; GP 1.5％ cream can inhibit UVB-induced activation of NF-κB and p38MAPK pathways,which may be one of the mechanisms underlying its protective effects against inflammation and photodamage.