| Objective: The laryngeal carcinoma is the malignant tumor which endangers mankind's health and life,Laryngeal carcinoma with surgical operation and put to treat for mainly cure means, But after operation sufferer's larynx lose original pronunciation function, still very great patient's existence quality and existence rate of the degree influence laryngeal carcinoma.In recent years along with the exaltation of chemotherapy level, the chemotherapy occupies an important position more and more in the comprehensive treatment of laryngeal carcinoma, for example laryngeal carcinoma of new assistance chemotherapy, its purpose lies in contract tumor and make clinical stage in advance, with radiotherapy or the partial laryngectomy combine to reserve a larynx voiced function. Laryngeal carcinoma because of position concealment, discover usually already arrive medium advanced stage, advanced stage laryngeal carcinoma or through radiotherapy or after partial laryngectomy again the surgical operation is more difficult, the simplicity radiotherapy to aim at local lesion, and hard to receive ideal result, clinically give chemotherapy, to control tumor of the whole body transfer, alleviating symptom, prolonging life.Laryngeal carcinoma With platinum is basic medication at head the neck cancer under chemotherapy treatment. With cisplatin (cDDP) for basic chemotherapy at head neck tumor the under medical treatment study more, and obtained certain curative effect, because of agreeable kidney toxicity and digest way side effect of cisplatin severity limited it in the clinical application,So develop efficiently low toxical platinum product has been the direction that the scholars study.Lobaplatin is the third generation platinum derivatives after cisplatin and carboplatin, the lobaplatin with its lighter side effect, with cross to drug fast incompletely, and stronger anti-tumor activity already initial show its important function in tumor under medical treatment.This experiment take the laryngeal carcinoma cell line Hep-2 as model, adopt MTT chromatometry and inverted -microscope and flow-cytometry to examine lobaplatinto to the Laryngeal carcinoma cell multiplication, cycle blockage,induce cell apopotosis etc.apply immunocy tochemical method to detect the expression of apoptosis related egg white bcl-2. Initially approach the effect lobaplatin to the Laryngeal carcino -ma cell multiplication, cycle blockage, induce cell apopotosis and the expression of apoptosis related egg white bcl-2. For lobaplatin can used for head and neck tumor ,s chemotherapy provide experiment basis.Methods:1 The laryngeal carcinoma cell line Hep-2 was was diviced into 3 groups:①blank contral(culture fluid);②negative control(cell+culture fluid);③experiment group (cell+ lobaplatin + culture fluid); each group cultivate 24, 48, 72 hours respectively2 To observe the cell growth and the change of cell morphology by inverted- microscope every day.3 Examine lobaplatin to repress the generation of the laryngeal carcinoma cell by MTT assay.4 The change of cell apoptosis and cell cycle was analyzed by the folw cytometry.5 Use the immunity cell chemistry method examine the express -ion of egg whites bcl-2.6 Statistic methods: The quantitative data were performed with one-way-ANOVA and the two sample data was performed with t-test. P<0.05 were considered statistically significant.Results:1 The observe results of cell morphology: negative control Hep-2 cell as endepidermis, adherence growth in nutritive medium, polygon,Fusiform shape ,edge sharpness,cell structue is closed, growth productive.after 5μmol/L effect 48h, a small quantity cell changes a circle, still adherence growth, along with giveing the aggrandizement o fdose of lobaplatin, become round cell amount to increase, arrive 80μmol/L, greatly parts of cells suspend in the nutritive medium. Becoming round cell to increase obviously is all suspended in nutritive medium, and appear a lot of fragments around the cell.2 The effect of lobaplatin to cell multiplication: Different dose lobaplatin at same time A value and matched control comparison, 5μmol/L function 24 hours and matched control comparison, the difference was not statistically significan (p>0.05),rest each group and matched control comparison, the difference all were statistically significan (p<0.05).Same dose lobaplatin function 24h,48h,72hcompare difference to all have statistically significan (p<0.05).The lobaplatin can repress the laryngeal carcinoma cell to propagate obviously, inhibition ratio rise along with the increment of dose of lobaplatin, having dose-dependent relation; inhibition ratio rise along with medicine function time of extension, having time-dependent relation. But arriving certain medicine dose and time ,increase again the lobaplatin dose, inhibition ratio increment was not obviously, presenting a platform effect.3 The influence of lobaplatin to cell cycle:Same dose of lobaplatin effect 24, 48, 72 hours S cycle and the G0/G1cycle all have statistical significance compare to control group (p<0.05). 5μmol/L lobaplatin start causing the laryngeal carcinoma cell'S cycle proportion increment for 24 hours, prolonging to 72 hours the S cycle proportion to descend, G0/G1 cycle cell proportion increment.20μmol/L lobaplatin make cell hold-up in S cycle for 24 hours,along with time extension, the S cycle proportion to descend, appearing obvious G0/G1 blockage later.Make while expressing low dose in a short time make cell hold-up in S cycle , high dose in a long time make cell hold-up in G0/G1 cycle.4 Lobaplatin influence to the cell apoptosis rate:The normal control cell has no apoptotic peak,5μmol/L lobaplatin appear small apoptotic peak after influence to the cell for 24 hours, appear obvious apoptotic peak after influence to the cell for 72 hours, 20μmol/L lobaplatin appear obvious apoptotic peak after influence to the cell for 24,48 hours.Different dose lobaplatin influence to laryngeal carcinoma cell 24,48,72 hours the difference all haave statistical significance (p<0.05), same dose lobaplatin influence to laryngeal carcinoma cell 24, 48,72 hours the difference all have statistical significance (p<0.05).The result expresses the lobaplatin can induce the laryngeal carcinoma cell apoptosis,and along with the increment of the medicine dose and time, the apoptosis rate increases gradually, the lobaplatin induce laryngeal carcinoma cell apoptosis of the time and dose-dependence relation,low dose lobaplatin (5μmol/L) can induce the laryngeal carcinoma cell apoptosis for 24 hours.5 Adopt immunocy tochemical method(S-P method) examines the expression of the egg whites bcl-2 in each group, the expressions of bcl-2 die down after 20μmol/L lobaplatin function 48hs,by Student's t-test of significance,p<0.05,the difference has statistical significanceConclusion: 1 The lobaplatin can repress the laryngeal carcinoma cell to propagate obviously in vitro. having dose and time-dependent relation.2 Lobaplatin can make the laryngeal carcinoma cell cycle blockage, low dose in a short time make cell hold-up in S cycle,high dose in a long time make cell hold-up in G0/G1 cycle.3 The lobaplatin can induce laryngeal carcinoma cell apoptosis,it is of the time and dose-dependence relation,low dose lobaplatin (5μmol/L) can induce the laryngeal carcinoma cell apoptosis for 24 hours.4 Lobaplatin may be pass adjust the expression of bcl-2, induce laryngeal carcinoma cell apoptosis, this may be the function mechanism that lobaplatin induce cell apoptosis.
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