| Objective: Lung cancer is a cancer which often occurs in malignanttumors.In recent years, lung cancer has become the leading cause of death inthe malignant disease because the increased morbidity and mortality year byyear. As the early symptoms of lung cancer are not typically and the lungcancer is prone to metastasis, it is already advanced, unresectable and treatedwith the integrated approach to radiation therapy, chemotherapy and othertreatment, when the lung cancer is diagnosed. However, the treatment effect isnot satisfactory. Invasion and metastasis has been one of the major causes ofdeath in patients with lung cancer.The domestic and foreign studies have shown that, EGFR is usuallyoverexpressed in malignant epithelial tumor cells and the level of expression issignificantly correlated with tumor prognosis and high expression predictedpoor prognosis. EGFR is one of epidermal growth factor receptor familymembers. The intracellular region contains a tyrosine kinase segment andligand binding sites. The tyrosine kinase activity is crucial in regulating cellproliferation and differentiation. The binding of ligand and EGFR caused theincrease in the level of gene transcription in the nucleus, the cell proliferation,and transformation and malignant through a series of signal transductioncascade. In addition, many domestic and foreign researches found that EGFRand COX-2have collaborative relationship in the occurrence and thedevelopment of lung cancer.The oncogene C-erbB-2encoded protein product is a tran-membraneprotein receptor which is also the member of the epidermal growth factorreceptor family as EGFR and is over expressed in many common tumors. Thestructure of the C-erbB-2protein is highly homologous with EGFR, so theC-erbB-2protein is not alone in lung cancer metastasis. The study found that the C-erbB-2protein often form dimeric with other members of theepidermal growth factor receptor family to promote cell growth andtransformation so as to promote the transfer of lung cancer.CXCR4is a chemokine receptor. It play an important role in the invasionof lung cancer metastasis when it combined with its ligand SDF-1. Manystudies have shown that tumor metastasis and inflammatory cell infiltration issimilar such as cells involves scrolling adhesion and transendothelialmigration. The interaction of CXCR4and SDF-1makes the tumor cells easilydetach from the primary site, invasion of human extracellular matrix andblood vessels.β-catenin is a multifunctional protein located in the cytoplasmic. It canmediate cell adhesion and involved in cell proliferation, differentiation andapoptosis. The study shows that the abnormal expression of β-catenin maylead to the activation of cell signaling pathways, and the activity of genetranscription in the nucleus. The activity improves the ability of cancermetastasis, thus contributing to the occurrence of tumor metastasis.Nimesulide is COX-2selective inhibitors, which has a preventive role inthe occurrence and growth of the tumor and tumor metastasis certain extent. Italso can enhance the effect of chemoradiotherapy. Oxaliplatin is a thirdgeneration platinum broad-spectrum anticancer drugs. It can inhibit DNAsynthesis and replication by generating alkalized conjugate role in the DNA,forming intra-chain and inter-chain cross linking. In this study, we determinethe level of EGFR, C-erbB2, CXCR4, β-catenin expression in tumor tissue byimmunohistochemical staining and fluorescence quantitative real-time PCRmethod.Then study the mechanism of tumor metastasis.Methods:26nude mice,4-5-week-old, BALB/c, male, weighing20~24grams, fed with sterile feed and sterile purified water in a rearingenvironment with ambient temperature, suitable humidity and no specificpathogens. With F12k medium containing12%newborn bovine serum,150ulpenicillin and300ul streptomycin, human lung cancer A549cells arecultivated in an incubator of37℃with5%C02and cells are digested and passed with0.25%trypsin when cells grow adherently to70%~80%confluence. Digest the A549cells in logarithmic growth phase with0.25%trypsin and dilute them with serum-free F12k dilution medium to a celldensity of1×10~7/ml after counted with CBC board. The human lung cancerA549cell suspension was injected subcutaneously in nude mice to establishaxillary tumor model. After the tumor diameter reached an average of4mm,removed the nude mouse which had the smallest tumor volume and thenrandomly divided them into four groups,6nude mice each group: controlgroup, nimesulide-treated group, oxaliplatin-treated group, and nimesulidecombined with oxaliplatin-treated group. Medecines were administeredrespectively. Nimesulide group by intragastric administration, dose of2.5mg/kg, administered1times daily; oxaliplatin group by intraperitonealinjection, a dose of10mg/kg/, administered2times a week. The control groupwere injected with saline, oral sterile distilled water.Execute nude mice afteradministering42days, cut and transplant tumor tissue, immunohistochemistrydetection of EGFR, C-erbB-2, β-catenin, CXCR4protein expressions,RT-PCR assay Tumor EGFR, C-erbB-2, β-catenin, CXCR4mRNAexpressions.Results: We performed immunohistochemistry analysis of EGFR,C-erbB-2, CXCR4, β-catenin protein in tumor samples. Analysis results showthat, compared with the control group, nimesulide group and combinationgroup, EGFR, C-erbB2, CXCR4and β-catenin expression levels weresignificantly lower (P <0.05); compared with the control group, oxaliplatingroup C-erbB2, the expression level of CXCR4had little change (P>0.05),while EGFR, β-catenin expression decreased (P<0.05); compared withnimesulide group, combination group, EGFR, C-erbB2, CXCR4and β-catenin protein expression levels were decreased (P<0.05). Analysis resultsshow that, compared with the control group, nimesulide group andcombination group, EGFR, C-erbB2, CXCR4and β-catenin expression levelswere significantly lower (P <0.05); compared with the control group,oxaliplatin group C-erbB2, the expression level of CXCR4had little change (P>0.05), while EGFR, β-catenin expression decreased (P <0.05);compared with nimesulide group, combination group, EGFR, C-erbB2,CXCR4and β-catenin protein expression levels were decreased (P <0.05).Determination of the expression of EGFR, C-erbB2, CXCR4, β-cateninmRNA real-time fluorescence quantitative PCR method, statistical analysisresults show that, compared with the control group, the expression ofnimesulide group and combination group, EGFR, C-erbB2, CXCR4, β-catenin levels of mRNA were decreased (P <0.05); compared with thecontrol group, the expression of EGFR, oxaliplatin group beta-catenin mRNAlevels decreased (P <0.05), while C-erbB2, CXCR4, mRNA levels showedlittle change (P>0.05); compared with nimesulide group, combination group,EGFR, C-erbB2, CXCR4, β-catenin expression of mRNA decreased (averagewater respectively P <0.05).Conclusion: Expression of a single application or application ofnimesulide combined with oxaliplatin could inhibit human lung cancerxenografts in nude mice of EGFR, C-erbB2, CXCR4, β-catenin. Oxaliplatincould inhibit the expression of EGFR, β-catenin. Combined use of nimesulideand oxaliplatin enhances the antitumor effect of oxaliplatin. |
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