Objective To investigate the expression changes of nuclear factor kappa B (NF-κB) and matrix metalloproteinase-9 (MMP-9) in the cultured hepatocellular carcinoma cells 9204 (HCC9204) transfected with inhibitory kappa B alpha（IκB-α)vector. Methods After pcDNA3-IκB-α vector and pcDNA3 were transfected into HCC9204 by lipofectamine method, Western-blot and RT-PCR analysis were used to detect the expressions of NF-κB and MMP-9. Migration and invasion of tumor cells were assayed by fundus membrane invaded by them. Results When pcDNA3-IκB-α was transfected into HCC9204, the expression of NF-κB was decreased at the protein level, and the expression of MMP-9 mRNA and the invision and metastasis ability of transfected cells were obviously decreased. Conclusion When the activity of NF-κB is inhibited, the ability of invasion and metastasis in HCC9204 cells decrease, which could be related to the decreased the expression of MMP-9.
ObjectiveTo study the role of survivin gene in the research work of tumor. MethodsLiteratures about the molecular structure, function, mechanism,distribution of survivin gene and its role in the treatment of tumor were reviewed.ResultsSurvivin was a new member of inhibitor of apoptosis protein family, expressed in almost all the human tumors independent to the expression of bcl2. The expression of survivin was significantly correlated with the poor prognosis of tumor patients. Survivin inhibited the apoptosis of tumor cells via inhibiting the activity of caspase3, the effector molecule of the apoptosis signal transduction pathway. Inhibition of the expression of survivin gene could block its effect of inhibiting apoptosis and consequently get a therapeutic effect of tumors.ConclusionSurvivin is commonly expressed in human tumor tissues. It can be identified as an important prognostic parameter of poor outcome and a new therapeutic target in cancer.
Objective To evaluate the effect of recombinant human growth hormone(rhGH) on growth of human colonic cancer cells (COLO-320) in vitro. Methods Human COLO-320 cells in logarithm growing period were cultured for 24 h,48 h or 72 h with variant concentrations of rhGH,camptothecine (CPT) or rhGH combined CPT in calf serum(serum group) or calf serum-free (serum-free group). Light density of cells were determined by MTT method, so that cellular inhibition rate were calculated.Results No influence on cell growth or inhibition rate was observed from cultures with variant concentrations and different acting times of rhGH (P＞0.05). Inhibition rate of single CPT or CPT combined rhGH were much more increased than single rhGH used (P＜0.01) with no statistical significance (P＞0.05).Conclusion The results show that rhGH has neither direct COLO-320 cells stimulation nor any evidence of COLO-320 cells inhibition, and has no influence of CPT on COLO-320 cells inhibition in vitro.
Objective To study the effect of p38MAPK activity on tumor necrosis factor-α （TNF-α） mRNA and intercellular adhesion molecule 1 （ICAM1） mRNA expressions of isolated rabbit liver during early stage of cold preservation and reperfusion period. Methods Based on the cold preservation and reperfusion model of isolated rabbit liver, the animals were divided into inhibition group （n=12） with 3 μmol/L SB202190 (p38MAPK specificity inhibitor) in perfusate and control group （n=12） with no SB202190 in perfusate. Liver tissue samples were harvested at the time points of before resection, end of cold preservation, and different reperfusion period (10, 30, 60 and 120 min). Protein expression and activity of p38MAPK were detected by Western blot and immunoprecipitation respectively, expression of TNF-α mRNA was detected by RT-PCR, and expression of ICAM1 mRNA was detected by in situ hybridization. Results There was no obvious change of expression of p38MAPK protein in liver tissue both in two groups during the total period （P＞0.05）， and there was no statistically significant difference between two groups （P＞0.05）. At time points of end of cold preservation, 10, 30 and 60 min of reperfusion, the activity of p38MAPK in control group was significantly higher than that at the time points of before resection and 120 min of reperfusion (P＜0.01), and was also significantly higher than that in inhibition group at the same time points (P＜0.01). There was no significant difference in activity of p38MAPK among all time points in inhibition group (P＞0.05). The expressions of TNF-α mRNA and ICAM1 mRNA at the time points of before resection, end of cold preservation, and 10 and 30 min of reperfusion were significantly lower than those in 60 and 120 min of reperfusion in both two groups (P＜0.05, P＜0.01); The expressions of TNF-α mRNA and ICAM1 mRNA in inhibition group were significantly lower than those in control group at the time points of 60 and 120 min of reperfusion (P＜0.01). The activity of p38MAPK of liver tissue during cold preservation and reperfusion period was significantly correlated with the level of TNF-α mRNA and level of ICAM1 mRNA expression （r=0.996, P＜0.01； r=0.985, P＜0.01）. Conclusions These results suggest that p38MAPK pathway may regulate the expressions of TNF-α and ICAM1 at the level of transcription and the activation of p38MAPK can up-regulate TNF-α and ICAM1 expressions, which may be one of the important mechanisms to cause ischemia-reperfusion injury of isolated liver during cold preservation and reperfusion period.
【Abstract】Objective To investigate the effects of selective cyclooxygenase-2 (COX-2) inhibitor nimesulide on the proliferation of colon adenocarcinoma cells in vitro and the expression of matrix metalloproteinase-2 (MMP-2). Methods The human colon cancer cell lines HT-29 and HCT-116 were employed in the study, grouped as nimesulide group, DMSO control group and blank control group. After treatment with nimesulide, the inhibitory effect of nimesulide on the proliferation of cancer cells was quantified by MTT assay, and the expression of MMP-2 in the cells was detected by quantitative zymography. Results Nimesulide inhibited the proliferation of HT-29 and HCT-116 cells in time and dosedependent manners. The inhibitory effect on HT-29 cells was ber than that on HCT-116 cells. Nimesulide downregulated the MMP-2 expression in HT-29 cells, whereas the expression in HCT-116 cells remained unchanged. Conclusion Nimesulide can obviously inhibit the growth of colon cancer HT-29 cells with positive COX-2 protein, suggesting that nimesulide may downregulate the expression of MMP-2 by inhibiting the activity of COX-2.
Objectives To investigate the expressions and significance of E2F1, ID1, and Bax protein in gallbladder adenocarcinoma tissues. MethodsThe expressions of E2F1, ID1, and Bax protein in 70 cases of gallbladder adenocarcinoma, 20 cases of high level intraepithelial neoplasia, 30 cases of low level intraepithelial neoplasia, and 20 cases of cholecystitis tissues were tested by using immunohistochemical method. ResultsThe positive expression rates of E2F1, ID1, and Bax protein in gallbladder adenocarcinoma was 84.3%, 70.0%, and 25.7%, respectively; the positive expression rates in high level intraepithelial neoplasia was 75.0%, 65.0%, and 55.0%, respectively; the positive expression rates in low level intraepithelial neoplasia was 16.7%, 23.3%, and 56.7%, respectively; and the positive expression rates in cholecystitis tissues was 10.0%, 20.0%, and 75%, respectively.The positive expression rates of E2F1 and ID1 protein in gallbladder adenocarcinoma were significantly higher than those intraepithelial neoplasia and cholecystitis tissues (P < 0.05), but the positive expression rate of Bax protein in gallbladder adenocarcinoma was lower (P < 0.05).The expressions of E2F1 and ID1 protein were significantly correlated with clinical Nevin staging of gallbladder adenocarcinoma (P < 0.05), but not correlated with the gallbladder adenocarcinoma differentiation degree (P > 0.05).The expression of Bax protein was related to the gallbladder adenocarcinoma differentiation degree (P < 0.05), but not correlated with clinical Nevin staging (P > 0.05).The expression of E2F1 protein was negatively correlated with expression of Bax protein (r=-0.375, P < 0.05), ID1 protein expression has nothing to do with the protein expression of Bax protein (P > 0.05).The expression of E2F1 protein was positively correlated with ID1 protein (r=7.031, P < 0.05). ConclusionsThe E2F1, ID1, and Bax may play an important role in the generation and development of the gallbladder adenocarcinoma.The combined detection of E2F1, ID1, and Bax have important guiding significance for auxiliary diagnosis and clinical staging of gallbladder adenocarcinoma.
Objective To review researches of the role of inhibitorof differentiation 2(Id2) in skeletal muscle regeneration. Methods The latest original literature concerning Id2 and its role in skeletal muscle regeneration was extensively reviewed. Results Id2 could form heterodimers by combining with E protein to prevent myogenic regulatory factors (MRFs) forming heterodimers by combining with E protein, to inhibit the transcription activity of MRFs anddifferentiation of skeletal muscle cell. Conclusion Id2 plays an important role in skeletal muscle regeneration.
Behcet's Disease (BD) is a multisystem vasculitis characterized by disease alternated with recurrent episodes and remissions, involving genital, oral, ocular uvea, cutaneous, and articular manifestations. The nuclear factor (NF)-κB signaling pathway paly an important role in the BD progression. It encompasses diverse gene, protein, and cellular regulatory mechanisms operating across various levels, alongside microbiological and experimental studies involving animals and cells. At the protein research findings, activation of the NF-κB pathway in BD patients is marked by elevated plasma levels of soluble CD40 ligand, which stimulates neutrophils to release reactive oxygen species and extracellular traps, thereby promoting inflammation. At the cellular research findings, macrophages in BD patients polarize towards classically activated macrophages phenotype through the NF-κB pathway, exacerbating the inflammatory response. The activation of NF-κB is associated with increased expression of anti-apoptotic proteins in T cells, leading to prolonged inflammation. Microbiological investigations reveal that the decreased gut microbiota diversity in BD patients compromises intestinal barrier integrity. NF-κB pathway involvement in regulating neutrophil and type 1 helper T cell (Th) 1/Th17 cell function worsens inflammation. Genetically, BD patients exhibit polymorphisms in immune regulatory genes, which contribute to inflammation through the NF-κB pathway. Mutations in NF-κB-associated genes elevate the risk of BD, while mutations in the endogenous inhibitor A20 lead to abnormal NF-κB activity, sustaining inflammation. Animal experiments and in vitro experiments corroborate the efficacy of NF-κB inhibitors in attenuating inflammation. Targeting upstream inflammatory factors within the NF-κB pathway yields positive outcomes in BD patients. In summary, the NF-κB signaling pathway plays a pivotal role in the development of BD. Developing NF-κB inhibitors may open new avenues for treating BD. Further research is necessary to comprehensively elucidate the precise mechanisms by which NF-κB operates in the pathogenesis of BD, as well as its potential clinical applications in therapy.
ObjectiveTo detecte the expressions of phosphatase and tensin homolog deleted on chromosome ten (PTEN) gene and protein in peripheral blood mononuclear cells (PBMC) of patients with sepsis and to explore its role in the pathogenesis of acute obstructive suppurative cholangitis (AOSC). MethodsThe PBMC and serum were separated from AOSC patients (n=25) before treatment and in 1 week after recure, and healthy volunteers (normal control group, n=15). The serum levels of lipopolysaccharide (LPS), tumor necrosis factor-α (TNF-α) and interleukin 10 (IL-10) were detected by enzyme linked immunosorbent assay (ELISA) methods. The mRNA and protein expression levels of PTEN, nuclear fator κB P65 (NF-κB), and inhibitor of NF-κB (IκB) were detected by real-time quantitative polymerase chain reaction (qRT-PCR) and Western blot, respectively. ResultsThe levels of LPS, TNF-α, and IL-10 before treatment were significantly higher than those of normal control group (P < 0.05), the indicators were significantly decreased and close to normal levels in 1 week after recure. The mRNA and protein expression levels of PTEN and IκB before treatment were lower than those of normal control group (P < 0.05), NF-κB P65 was higher than those of normal control group (P < 0.05), while the phosphorylation levels of PTEN and IκB were higher than those normal control group (P < 0.05), and in 1 week after recure, the above indicators returned to normal levels. ConclusionsSepsis shift may be associated with the occurrence of intestinal LPS, and caused the imbalance between proinflammatory and anti-inflammatory cytokines in the body. PTEN for phosphorylation activation of IκB or directly activation of NF-κB participate the originating process of sepsis, hinting a therapeutic potentialities in the early stage of sepsis.
ObjectiveTo observe RNA-Seq analysis of gene expression profiling in human retinal vascular endothelial cells after anti-vascular endothecial growth factor (VEGF) treatment.MethodsCultured the retinal vascular endothelial cells in vitro and logarithmic growth phase cells were used for experiments. The cells were divided into VEGF group and VEGF combined with anti-VEGF drugs group. The VEGF group cells were treated with 50 ng/ml VEGF for 72 h to simulate the high VEGF survival conditions of vascular endothelial cells in diabetic retinopathy. VEGF combined with anti-VEGF drug group cells was treated with 50 ng/ml VEGF and 2.5 μg/ml anti-VEGF drugs for 72 h to imitate the microenvironment of cells following the anti-VEGF drugs treatment, and whole transcriptome sequencing approach was applied to the above two groups of cells through RNA-Seq. Now with biological big data obtained as a basis, to analyze the differentially expressed genes (DEGs). And through enrichment analysis to explain the differential functions of DEGs and their signal pathways.ResultsThe gene expression profiles of the two groups of cells were obtained. Through analysis, 328 DEGs were found, including 194 upregulated and 133 downregulated ones. The functions of DEGs were influenced by regulations over molecular biological process, cellular energy metabolism and protein synthesis, etc. Among these genes, SI,PRX and HPGD were related to protein synthesis, BIRCT to cellular apoptosis, and ABLIM1 and CRB2 to retinal development, and ABCG1, ABCA9 and ABCA12 were associated with the cholesterol of macrophage and the transfer of phospholipid. GO enrichment analysis showed that DEGs mainly act in three ways: regulating biological behavior, organizing cellular component and performing molecular function. Pathway enrichment analysis showed that gene expressions of the two cell groups were differentiated in ECM receptor pathway, and Notch, mitogen-activated protein kinase, transforming growth factor (TGF)-β and Wnt signal pathways. Among them, the gene expression in TGF-β signal pathway attracts most attention, where the DEGs, such as CAMK2B, COL3A1, CYGB, PTGER2 and HS6ST2, among others, were closely related to fibrosis process.ConclusionThe anti-VEGF drugs may enhance the expression of CAMK2B, COL3A1, CYGB, PTGER2 and others genes related to TGF-β signal pathway and aggravate retinal fibrosis disease.