| objective: To investigate the effect and clinicopathologicalsignificance of RECK gene in lymphatic metastasis of oral squamouscarcinoma,explore the regular mechanism of RECK gene in lymphaticmetastasis of oral squamous cell carcinoma and study the impossible actionmechanism of Art to anti-invasion and anti-metastasis in oral squamous cellcarcinoma.Methods:1. To collect clinical specimens of oral squamous cellcarcinoma60cases with experimental conditions, using immunohistochemicalmethod to detect RECK gene and MMP-9in tumor tissues and analyze therelationship between the expressionof RECK and MMP-9and clinicalsignificance, and analyzy the mutual relations between RECK and MMP-9;2.Human oral squamous cell carcinoma Tca8113cells in vitro, with differentconcentrations of ART intervention, using MTT assay the growth inhibition ofeach concentration group Tca8113cell and flow cytometry analyzes theinfluence of apoptosis and cell cycle distribution of various concentrations ARTon Tca8113cell; immunohistochemical method detect the influence of RECKprotein and MMP-9protein expression of various concentration ART onTca8113cell;3. Through the establishment of lymphatic metastasis of oralsquamous cell carcinoma in animal models, and with artesunate experimentaltherapeutic interventions, using immunohistochemical analyze RECK gene and MMP-9expression in the tumor and lymph node metastasis, analyze artesunateintervention impact on inhibition of oral squamous cell carcinomametastasis.Results:1.60cases clinical specimens found expression of RECKgene and MMP-9are closely related to lymph node metastasis in oral squamouscell carcinoma and malignant degree (P<0.05), with the malignant degree ofincrease, degree of lymph node metastasis of progressive, RECK expressionlevels gradually decline and showing negative correlation with the expressionlevel of MMP-9(P<0.05);2. ART intervention of Tca8113cells in vitroexperiment found that ART can effectively inhibit Tca8113cell proliferationand induction of apoptosis in early and late, and early apoptosis rate greaterthan late apoptosis rate, as the intervening time increase, extension of its role inthe concentration of ART is more and more obvious, and in72h the mostobvious. Immune group of results displayed, ART roled72h, as medicationconcentration increased, the number of the cell with RECK positive expressionis gradually increased, its average light density value also gradually rise, and inconcentration for200ug/ml is highest, the differences of all concentration groupand control group is statistics significance (P<0.05), and the number of the cellwith MMP-9positive expression except12.5ug/ml, remaining as drugconcentration increased, its light density value gradually reduced, and in200ug/ml is the minimum,the difference of various concentration comparedwith the control group is statistically significant (P<0.05);3. Animal model experiment found that after the intervention of ART therapy in200mg/kgconcentration, an average weight of the transplanted tumor in ART group issignificantly less than the negative control group (P<0.05), showing that ARTcan significantly inhibit growth and invasion and metastasis of oral squamouscell carcinoma, lymph node metastasis of oral squamous cell carcinoma inprotein expression of RECK and MMP-9, the difference was not statisticallysignificant compared to group (P>0.05).Conclusion:1. RECK gene proteinlevels in oral squamous cell carcinoma was negatively correlated with oralsquamous cell carcinoma’s pathological grading, TNM stage and lymph nodemetastasis,but it does not matter with the clinical characteristics of the patient’sage, gender and tumor size, but MMP-9’protein level was gradually increase.AsClinical and pathological grading, TNM staging, lymph node metastasis is themain index to determine the prognosis of cancer patients, therefore, thedetection of RECK gene protein expression levels may be the prognosis ofpatients with a certain reference value;2. MMP-9and RECK gene expression inoral squamous cell carcinoma is closely related to clinicopathological featuresof patients with oral squamous cell carcinoma, we can rise RECK geneprotein expression to cut MMP-9protein expression to suppress the invasionand metastasis of oral squamous cell carcinoma；3. Tca8113ART can inhibitoral squamous cell carcinoma proliferation, and promoted effects to theapoptosis in early and late-stage, as well as the concentration increases gradually increased;4. ART can increase expression of RECK gene throughcut MMP-9protein expression in Tca8113cell to inhibit oral squamous cellcarcinoma proliferation,and promoted effects to the apoptosis;5. ART caninhibit the growth, invasion and lymph node metastasis of oral squamous cellcarcinoma, which is prompted ART in the treatment of oral squamous cellcarcinoma has a potential value of basic and clinical research, and provides adirection for new adjuvant chemotherapy anticancer drugs and drug screening;... |
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