| [Objective]:Detection Xuanwei area non-small cell lung cancer patients EML4-ALK fusion gene, and explore different concentrations Tae684(ALK inhibitor) on XWLC-05, BEAS-2B cell growth.[Method]:1. Select the March2011to April2012Third Affiliated Hospital of Kunming g Medical University, thoracic surgery postoperative lung cancer diagnosed54cases. Sherwin region where27cases,27cases of non Sherwin region, the collected specimens by immunohistochemistry method of specimens ALK-related protein detection.2. pairs of lung cancer patients who tested positive for a summary of the clinical features, the expression on the situation around to compare.3. using tetrazolium blue (MTT) method for the determination of different times and different concentrations Tae684and CDDP on XWLC-05cells, BEAS-2B cell growth, cells were divided into four groups and each negative control group, solvent control group, the experimental group (join Tae684) and positive control group (join CDDP), determine the median inhibitory concentration degrees (IC50);4. Flow cytometry was used to detect XWLC-05and BEAS-2B, respectively, in Tae684and CDDP for24h and48h, to detect the cell cycle and cell apoptosis.5. XWLC-05cells were treated with tae-684and CDDP-induced72h, fixative treatment, immunofluorescence microscopic fibers observed apoptotic bodies;6. Statistics and analyzed using SPSS17.0software package. Data were expressed as mean±standard deviation (±s) that the groups using single factor analysis of variance data (ONE WAY ANOVA) and t test.[Results]:1.27cases of non-small cell lung cancer Xuanwei in4cases were positive,27cases of other parts of Yunnan, non-small cell lung cancer in two cases expressed as positive. Cases detected in Xuanwei expression was14.8%; non Sherwin region of7.4%. Two regions were compared between groups p=0.669>0.05, the difference was not statistically significant..2. pairs of54cases of lung cancer in Yunnan postoperative specimens for immunohistochemical detection of ALK, six cases were positive expression rate was11.1%. Four cases in which strong positive yang, positive there are two cases.5cases of adenocarcinoma, large cell carcinoma. ALK (+) of the patients, through the comparison between groups in hemoptysis, lymph node metastasis, p-value<0.05, the difference was statistically significant.3. Tae-684and CDDP on XWLC-05cells BEAS-2B cells in vitro inhibition with time and concentration, the inhibition rate increases, XWLC-05cells was6.47±0.78μg/ml in Tae684and CDDP4.78±1.80μg/ml, BEAS-2B cells Tae684was4.46±1.16μg/ml and CDDP2.13±0.20μg/ml. In the inhibition rate, four groups through the comparison between groups, p<0.05, statistically significant difference.4.3nM of tae684right BEAS-2B cell cycle arrest occurs when24h, G2phase cells gradually how much,48h G2phase cells reached73.6%; apoptosis detection is gradually increasing,48h BEAS-2B apoptosis rate14.5%.3nM of tae684right XWLC-05cells were apparent at24h cycle arrest, G2phase cells gradually increased,48h G2phase cells reached84.3%; apoptosis detection is gradually increasing,48h XWLC-05cell apoptosis rate was 26.3%. By group comparison between the two groups, p<0.05, the difference was statistically significant.5. Tae-684-induced XWLC-05cells BEAS-2B cells after72h treatment by PI staining, fluorescence microscopy, in order to induce cell to produce apoptotic bodies dominated, leading to apoptosis.[Conclusion]:1. in Xuanwei region of ALK non-small cell lung cancer-related protein expression rate similar to other areas in Yunnan. From the expression, the Yunnan region is slightly higher than the Sherwin region;2.non-small cell lung cancer in Yunnan Province in ALK (+) patients generally have hemoptysis, cut earlier onset of lymph node metastasis;3. Tae684to XWLC-05and BEAS-2B cells in a certain range of dose and time dependent manner; Tae684right XWLC-05cell line is stronger than BEAS-2B cells;4. Tae-684to XWLC-05and BEAS-2B cells, to induce cells to produce mainly apoptotic bodies, leading to apoptosis. |
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