| [Objective] Detection of epidermal growth factor receptor expression in laryngeal cancer. Observation of gold nanorods and anti-epidermal growth factor receptor functionalized gold nanorods modified inhibition on cell value, and explore functional modification of GNR before and after the thermal effects on laryngeal carcinoma cell HEP-2 and lung epithelial cells BEAS-2B of anti- effect and apoptosis. Provide a basis for furthe experimental study.[Methods]1,Detected the expression by western-blot method of EGFR in laryngeal carcinoma cell HEP-2, normal laryngeal epithelium and laryngeal carcinoma.2,Characterized and synthetic GNRs and EGFR-mAb/GNRs.Cultured carcinoma cell HEP-2 and normal lung epithelial cell BEAS-2B in vitro. Logarithm flat growth to 96 orifice plate.Adds eventually concentration for (0.01-0.32 nM) GNR solution and the epidermal growth factor receptor (monoclonal antibody functionalized modifications gold nano great solution, the MTT method detection gold nano bars and epidermal growth factor receptor (monoclonal antibody functionalized modifications gold nanoparticles to HEP-2 laryngeal cancer sticks BEAS-2B lung cells and bronchial epithelial cell proliferation inhibition.3,The above the cells into the following group:(1). NC group (2). pure Light group (Light) (3).0.04 nM GNR group (4).0.04 nM EGFRmAb/GNR group,MTT assay the proliferation of gold nanorods and epidermal growth factor receptor monoclonal antibody functional modification of gold nanorods irradiation on the laryngeal carcinoma cells HEP-2 and bronchial epithelial cells in lung AEBE-2B in the 808nm wavelength near-infrared. And by flow cytometry (floweytometry, FCM) detection of thermal effects of apoptosis. MTT assay the proliferation of gold nanorods and epidermal growth factor receptor monoclonal antibody functional modification of gold nanorods in the 808nm wavelength near-infrared (near-infrared, NIR) irradiation on (0.8-4w/cm2) the HEP-2 laryngeal carcinoma cells and pulmonary BEAS-2B bronchial epithelial cell and detection of thermal effects of apoptosis by flow cytometry.4,Experimental results data is mean±standard deviation (x±s), using spss17.0 statistical software processing, two sets of data using the statistical analysis t test. Many sets of data in a single factor analysis of variance between two comparison between groups, the results are all take P<0.05 determined statistical significance.[Results]1,Western Blotting semi-quantitative results show that high EGFR expression in laryngeal carcinoma, and its expression is much higher than normal mucosa (P <0.05).2,The same gold particle size, distribution, and good monodisperse particles showed a rod-like, with an average aspect ratio of 3.9, the solution will not be stable for a long time placed. in the 540nm and 822nm vicinity of plasma resonance absorption have two peak, in the 808nm absorption peak vicinity of near-infrared region (800-1200nm) is the organization's security transmission window of the body, which can be used near infrared hyperthermia. With MTT cytotoxicity assay found the survival rate decreased with increasing concentration. Half of the inhibition rate of concentration of the drug (IC50), respectively, for the 0.07nM and 0.14nM (HEP-2),0.14nM and 0.18 nM (BEAS-2B cells).3,HEP-2 cells and BEAS-2B cells after the application of near infrared laser irradiation on cell proliferation and growth are inhibited; and the inhibition increased with the irradiation power, cell survival was also decreased. HEP-2 P values between the three groups were<0.01 for significant difference. Relative to the negative control group, all groups was statistically significant, P<0.05, BEAS-2B cells between the two groups,the P values were> 0.05, the difference was not statistically significant. HEP-2 cells and both BEAS-2B cells were treated the same as the role of near-infrared laser hyperthermia, BEAS-2B cells of each experimental group inhibited were far less than the inhibitory effect of HEP-2 cells.4,gold nanorods have NIR irradiation induced cell apoptosis in a dose dependent manner. The pure light group alone and without NIR irradiation of gold nanorods were not significantly induce apoptosis or promote cell death. only in the performance of high concentrations of 0.04 nM can has weak apoptosis, the apoptosis rate was 3.1±0.51%. Light group compared with the negative control group of gold nanorods has significant difference of apoptotic cells.[Conclusions]1. EGFR is highly expressed in carcinoma cell of laryngeal squamous, and its expression is much higher than normal mucosa.2. a large number of the gold nanorods in the form of nanoparticles and it enter into HEP-2 cells did not change the shape. A large number of gold nanorods present in the cytoplasm and mitochondria of cell. without affecting the cell state it can co-exist with cell in IC50 concentrations for a long time.3. Gold nanorods modifed by anti-epidermal growth factor receptor monoclonal antibody has good functional of directional aggregation to HEP-2 cells, compared with the gold nanorods it using low near-infrared laser power required for hyperthermia, HEP-2 laryngeal cancer cells more sensitive to heat than normal lung epithelial cells4.In near infrared laser irradiation gold nanorods can induced cell apoptosis in a dose dependent manner. Gold nanoparticles can induce tumor cell apoptosis in appropriate conditions... |
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