ObjectiveTo explore the relationship between nuclear factor κB (NFκB) and the occurrence, metastasis, and treatment of colon cancer. MethodsThe literature on the structure and the property of molecular biology of NFκB, the relationship between NFκB and apopotosis, malignant tumor and colon cancer were reviewed.ResultsNFκB had action of antiapopotosis. The occurrence of malignant tumor had close relation with the oncogene by NFκB, the metastasis of malignant tumor was that cell of cancer escaped the killing and supervising of immunity by NFκB. NFκB affected the occurrence and metastasis of colon cancer by regulating cmyc, Cox2, ICAM1.Conclusion NFκB has important action in the occurrence and metastasis of colon cancer. It will become a new target of treatment.
Objective To investigate the expression of transcription factors including nuclear factor-κB （NF-κB） and activator protein-1 （AP-1） in vascular endothelial cells （ECs） in different flow fields， and provide experimental evidence for mechanical signal effects on gene regulation pattern of ECs. Methods Cultured human umbilical vein ECs were loaded into steady flow chambers of laminar flow or turbulent flow and observed at 6 time points （0.5 h， 1 h， 2 h， 3 h， 4 h and 5 h） based on different load time. Spacial and temporal characteristics of NF-κB and AP-1 expression in ECs in different flow chambers were detected at a protein level by laser confocal microscope. Results In laminar flow， NF-κB expression rose to peak at 1 hour （26.49±1.63， P＜0.05）and then declined. In turbulent flow， NF-κB expression rose to peak at 3 hours （34.41±6.43， P＜0.05）. In laminar flow， c-Jun/AP-1 expression was transiently elevated， reached its peak at 0.5 hour （18.95±5.38，P＜0.05）and then fell to its baseline level. In turbulent flow， c-Jun/AP-1 expression rose slowly but steady to peak（P＜0.05） . Conclusion The effects of turbulent flow on NF-κB and AP-1 expression in ECs are different from those of laminar flow. Up-regulation and activation of NF-κB and AP-1 expression in ECs induced by turbulent flow may cause pathological changes in morphological structure and functional behavior of ECs.
Objective To construct the mouse NF-κB P65 subunit expression plasmid, and identify its biological activity. Methods NF-κB P65 siRNA retrovirus expression vectors were reconstructed by molecular clone technology. Recombinant vectors were transfected into 293E package cells and virus suspension was collected. RT-PCR was used to detect the expression level of NF-κB P65 mRNA and TNF-α mRNA at different time-point of LPS stimulation. Western blot was performed to analyze the protein level of NF-κB P65. ELISA was applied to detect the expression level of TNF-α released by LPS-stimulated J774A.1. Results NF-κB P65 siRNA retrovirus expression vectors of mouse were successfully constructed. From2 hours after the stimulation of LPS, the expression level of NF-κB P65 mRNA of the siRNA group was obviously lower than the scramble control group ( 0.91 ±0.03 vs. 1.02 ±0.02, Plt;0.01) . At24,36, 48 and 72 hours after the LPS stimulation, the expression level of NF-κB P65 protein of the siRNA group was significantly decreased compared with the scramble control group ( 0.97 ±0.02 vs. 1.01 ±0.01, 0.94 ± 0.01 vs. 1.02 ±0. 01,0.94 ±0.02 vs. 1.02 ±0.01, 0.93 ±0.01 vs. 1.00 ±0.02, Plt;0. 05) . At 2, 6, 12, 24 hours after the LPS stimulation, both the expression level of TNF-α mRNA and the content of TNF-α in the culture medium supernatant of the siRNA group were lower than the scramble control group ( Plt;0. 01) . Conclusions The construction of NF-κB P65 siRNA retrovirus expression vectors is feasible. Inflammation factors in mouse monocyte-macrophages are significantly inhibited after NF-κB expression is depressed by RNA interference technology, which may be applied to prevent and treat excessive inflammatory reaction in acute lung injury.
【 Abstract 】 Objective Overexpressions of epidermal growth factor (EGF) and EGF receptor have been associated with progression and invasive phenotype of pancreatic cancer. However, the underlying molecular mechanism by which EGF worked in pancreatic cancer cells has not been completely understood. In this study, effect of EGF on the invasion and metastasis of pancreatic cancer cells and its regulatory mechanism were investigated. Methods The effects of EGF on the proliferation, adhesion and invasion of pancreatic cancer cells were detected by WST-1 proliferation assay, adhesion assay and invasive assay, respectively. The activity and expression of MMP-2 and MMP-9 were examined by zymography, Western blot and RT-PCR, respectively. The activity of NF- κ B was examined by EMSA. Results EGF could significantly promote the invasiveness of pancreatic cancer cells but did not affect cell proliferation or adhesion. The expressions of NF- κ B and MMP-9 were significantly increased by EGF, but EGF did not affect the activity and expression of MMP-2. Furthermore, EGF stimulated the NF- κ B binding activity. Pretreatment with NF- κ B inhibitors, pyrrolidine dithiocarbamate (PDTC), could significantly inhibit the activity of NF- κ B induced by EGF. Meanwhile, the EGF-induced expression and activity of MMP-9, as well as cell invasiveness were also inhibited by NF- κ B inhibitor. Conclusion EGF could increase the expression and promote the invasiveness of MMP-9 via the activation of NF- κ B in pancreatic cancer cells, which implies that NF- κ B inhibitant, such as PDTC, may diminish the invasiveness of pancreatic cancer cells.
As an important intracellular genetic and regulatory center, the nucleus is not only a terminal effector of intracellular biochemical signals, but also has a significant impact on cell function and phenotype through direct or indirect regulation of nuclear mechanistic cues after the cell senses and responds to mechanical stimuli. The nucleus relies on chromatin-nuclear membrane-cytoskeleton infrastructure to couple signal transduction, and responds to these mechanical stimuli in the intracellular and extracellular physical microenvironments. Changes in the morphological structure of the nucleus are the most intuitive manifestation of this mechanical response cascades and are the basis for the direct response of the nucleus to mechanical stimuli. Based on such relationships of the nucleus with cell behavior and phenotype, abnormal nuclear morphological changes are widely used in clinical practice as disease diagnostic tools. This review article highlights the latest advances in how nuclear morphology responds and adapts to mechanical stimuli. Additionally, this article will shed light on the factors that mechanically regulate nuclear morphology as well as the tumor physio-pathological processes involved in nuclear morphology and the underlying mechanobiological mechanisms. It provides new insights into the mechanisms that nuclear mechanics regulates disease development and its use as a potential target for diagnosis and treatment.
Objective To observe the protective effect on rat lung by using N-acetyl-L-cysteine(NAC) a inhibiter of nuclear factor-kappa B (NF-κB) in the period of reperfusion. Methods Twenty-four rats were randomly divided into a control group and a trail group.The harvested lung blocks of 12 rats were flushed with and stored in the low-potassium-dextran (LPD) solution at 4℃ for 16 hours. The isolated rat lung reperfusion models were established and the donor lungs were perfused for 1 hour. NAC was used in the trail group but normal saline was used in control group. Partical pressure of oxygen in artery (PaO2), peak airway pressure (PawP) were measured at every 15 min intervals during reperfusion. After reperfusion, the lung tissue wet-to-dry(W/D)ratio, and myeloperoxidase(MPO) activity were obtained. The protein and mRNA expressions of intercellular adhesion molecule-1(ICAM-1), NF-κB were also observed by using immunohistochemistry and semi-quantitative RT-PCR at the end of reperfusion. Results The level of decreased PaO2 and increased PawP in trail group were lower than those in control group at every interval time the sample obtained after reperfusion in 60 min. (Plt;0.01 or lt;0.05). After reperfusion the W/D,MPO, the protein and mRNA expressions of ICAM-1, NF-κB were decreased evidently in trail group than those in control group(Plt;0.01 or lt;0.05). Conclusion Using NAC in the period of reperfusion, can effectively inhibit the expression of NF-κB and ICAM-1,further improve lung respiratory functions.
Objective To investigate the mechanism of radix salviae miltiorrhizae (RSM) injection in treatment of pancreatitis through observing the changes of activity of nuclear factor-kappa B (NF-κB) in pancreas of rats with severe acute pancreatitis (SAP) and the influence of RSM injection upon NF-κB in pancreas tissue. Methods Seventy-five rats were randomly divided into 3 groups: normal control group, SAP group and RSM treatment group, which were injected with normal saline, normal saline or RSM in the peritoneal cavity, respectively. The model of SAP rats was made by injecting L-arginine into peritoneal cavity and by subcutaneous injection at the same time. The concentrations of amylase in plasma and in ascites were measured respectively, and the expression of NF-κB in pancreas tissues was determined by immunohistochemistry. Results The levels of amylase in plasm and ascites in SAP group and RSM treatment group increased significantly with the increased expression of NF-κB in pancreas tissue, but it was also found that both the level of amylase and the expression of NF-κB in RSM treatment group were significantly lower compared with those in SAP group, and the survival time of RSM treatment group was longer than SAP group with less pathological injury in the pancreas tissues. Conclusion RSM may be effective for the treatment of pancreatitis by degrading the expression of NF-κB.
Objective To study the effect of nontypeable Hemophilus influenzae(NTHi) strain ATCC49247 on proinflammatory cytokines expression of human A549 lung epithelial cell line. Methods Confluent A549 cells were co-incubated with NTHi, NTHi+Erythromycin(10 mg/L), NTHi+Gentamicin(100 mg/L), and NTHi+Dexamethasone(100 μmol/L),and nuclear factor kappa B(NF-κB) inhibitor primed cells were co-incubated with NTHi for 24 h. Then levels of interleukin-8（IL-8） and tumor necrosis factor-α(TNF-α) in the supernatant was assayed by enzyme-linked immunosorbent assay(ELISA) and the expression of intercellular adhesion molecule-1(ICAM-1) in cells was detected by immunohistochemistry staining. Results A549 cells were transformed and died after co-intubated with NTHi for 24 h. NTHi induced A549 cells to release significantly greater amounts of IL-8, which was inhibited by NF-κB inhibitor pyrrolidine dithiocarbamate（PDTC）. Incubating of A549 cells with NTHi significantly induced release of IL-8 and the expression of ICAM-1, which was blocked by erythromycin and dexamethasone and not by gentamicin. TNF-α was not detected in all circumstances. Conclusions NTHi can increase significantly the release and expression of proinflammatory cytokines through NF-κB pathway. Antibacterial drug erythromycin also has anti-inflammatory effect.
ObjectiveTo investigate the relationship between alumina ceramic particles and aseptic loosening of the joint prosthesis and the effect of lanthanum chloride on the secretion of interleukin 1β (IL-1β) and tumor necrosis factor α (TNF-α) of macrophage RAW264.7 induced by alumina ceramic particles.
MethodsRAW264.7 cells were cultured in vitro and divided randomly into 4 groups according to different culture solutions:blank control group (group A),1 mg/mL alumina ceramic particles (group B),1 mg/mL alumina ceramic particles and 10 μmol/L lanthanum chloride (group C),and 10 μmol/L lanthanum chloride (group D).The cell growth was detected by MTT,and ELISA,RT-PCR,and Western blot were used to test the expressions of IL-1β,TNF-α,and nuclear factor κB (NF-κB).
ResultsThere was no significant difference in cell growth among all groups by MTT (F=2.180,P=0.142).RT-PCR results showed that the expressions of IL-1β,TNF-α,and NF-κB mRNA in group B were significantly higher than those in the other 3 groups (P<0.05); the expressions in group D were significantly lower than those in group A (P<0.05).ELISA results showed that the contents of IL-1β and TNF-α in group B were significantly higher than those in the other 3 groups (P<0.05); the contents in group D were significantly lower than those in group A (P<0.05).Western blot analysis revealed that the expression of NF-κB protein in group B was significantly higher than that in the other 3 groups (P<0.05).
ConclusionAlumina ceramic particles can stimulate the secretion of IL-1β and TNF-α of macrophage,and lanthanum chloride can inhibit the secretion of IL-1β and TNF-α of macrophage.
Objective To investigate the roles of NF-κB and EGFR in hepatolithiasis associated with intrahepatic cholangiocarcinoma. Methods Ninety cases of liver tissue specimens from hepatectomies performed in the 2nd Affiliated Hospital of Sun Yat-sen University between August 1989 and June 2009 were enrolled in the study. Among them, 33 cases of hepatolithiasis associated with intrahepatic cholangiocarcinoma were considered as observing group, 32 cases of hepatolithiasis as control group, and 25 cases of normal bile duct tissues as normal control group. The SP method of immunohistochemical staining was applied to detect the expressions of NF-κB and EGFR in intrahepatic biliary ducts epithelial cells, and their relations with clinicopathologic factors and the accumulated survival rate of hepatolithiasis associated with intrahepatic cholangiocarcinoma were analyzed. Results Expression rates of NF-κB and EGFR were gradually raised from normal control group, control group to observing group (Plt;0.01). Expression of EGFR in tumor patients was related to histopathologic differentiation grading and the depth of tumor invasion (Plt;0.05), but not to gender, age, or lymph node metastasis (Pgt;0.05)； there were no significant relationships between the expression of NF-κB and factors described above (Pgt;0.05). The survival rate of patients with tumor expressed EGFR was significantly lower than that of patients with tumor non-expressed EGFR (Plt;0.01). Conclusions NF-κB expression is in the early stage during intrahepatic cholangiocarcinoma genesis. NF-κB and EGFR play cooperating roles during hepatolithiasis carcinogenesis process. Over expression of EGFR is related with poor differentiation and prognosis of tumor.
