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The Initial Approach on Efect of Lobaplatin in Laryngeal Carcinoma Cell Line-Hep-2

Author: ChenWenLing
Tutor: ZhuYanBing
School: Hebei Medical University
Course: Oncology
Keywords: Lobaplatin Laryngeal Cell cycle Apoptosis bcl-2 Hep-2
CLC: R739.6
Type: Master's thesis
Year: 2008
Downloads: 83
Quote: 1
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Abstract

Objective: throat cancer is a malignant tumor of harm to human health and life, its primary surgery and radiotherapy treatment after surgery in patients with laryngeal lost its original pronunciation, significantly affect the quality of life and survival rate of laryngeal cancer patients. In recent years, with the improvement of the level of chemotherapy, chemotherapy in laryngeal cancer treatment increasingly occupy an important position, such as laryngeal cancer neoadjuvant chemotherapy, the aim is to shrink the tumor, so the earlier clinical stage, and radiotherapy or partial laryngectomy surgery combined with the pronunciation function as much as possible to retain throat. Laryngeal location hidden, found often advanced, advanced laryngeal or post-operative recurrence in patients re-operation more difficult, radiotherapy only for localized lesions, it is difficult to receive the desired results, giving the clinical chemotherapy to control systemic metastasis of the tumor, relieve symptoms and prolong life. Platinum class is the basis of the head and neck chemotherapy drug, cisplatin (cDDP) based chemotherapy in head and neck cancer therapy research more, and have achieved a certain effect, due to severe renal toxicity of cisplatin and gastrointestinal side effects. limits its clinical application, so the development of high efficiency and low toxicity of platinum preparations has been the direction of research scholars. Lobaplatin is a third-generation platinum derivative the its lighter, lobaplatin, toxic side effects, and cisplatin incomplete cross-resistance as well as a strong anti-tumor activity, has been preliminarily shows its important role in cancer therapy. In this experiment, the laryngeal carcinoma cell line Hep-2 model, using an inverted microscope, MTT colorimetric assay the flow cytometry lobaplatin of laryngeal cancer cell proliferation, cell cycle arrest, and induce apoptosis role of immune cells chemical assay of apoptosis-related protein bcl-2 expression, explore Los platinum laryngeal cancer cell proliferation inhibition, cell cycle changes, induction of apoptosis and apoptosis-related protein expression and other aspects of the role, for lobaplatin in Head and Neck Ministry tumor chemotherapy to provide experimental evidence. Method: 1 laryngeal cancer cells to logarithmic phase experiments, experimental groups: (1) the control group (culture solution); the ② negative control group (cell culture medium); ③ experimental group (different cell concentrations lobaplatin culture medium ); each group were cultured for 48 and 72 hours. 2 inverted microscope to observe changes in cell growth and cell morphology. 3 Application MTT colorimetric to detect lobaplatin of laryngeal cancer cell proliferation inhibition. 4 application flow cytometry cell cycle and apoptosis rate change. 5 Immunocytochemistry method (SP) detection of bcl-2 protein expression changes. 6 Statistical analysis: SPSS13.0 statistical software package for statistical analysis. The first test of normality in the data analysis, in line with the normal distribution of measurement data expressed as mean ± standard deviation (xs). Homogeneity of variance between the two groups first meet the homogeneity of variance using ANOVA analysis methods, such as significant differences, further LSD method for pairwise comparisons. Two sample comparison t test, a significant level of p <0.05 as statistically significant. Results: 1 changes daily observation of the control group and the experimental group of cell size and morphology under phase contrast microscope. The negative control group Hep-2 cells, epithelioid adherent growth in culture media, and outline a clear, close cell structure, vigorous growth. 5μmol / L for 48h, lobaplatin, seen after a small amount of cell volume increases, and round, still adherent growth. Adherent growing cells gradually reduced 20μmol / L, rounded cells gradually increased, a small ball of cells and cell debris floating off the wall gradually increased to 80μmol / L high concentration of drug action adherent growth cells rarely appeared piles dead cells and cell debris floating lobaplatin cell proliferation: different concentrations lobaplatin compared with the control group in the same time A value 5 μmol / L lobaplatin for 24 hours with the control group was not statistically significant, the rest of the group and The control group differences were statistically significant (p lt; 0.05). Same the concentration lobaplatin role 24,48,72 differences were statistically significant (p lt; 0.05). Lobaplatin can significantly inhibit the laryngeal carcinoma cell proliferation inhibition rate increased with the increase of drug concentration, dose-dependent relationship; inhibition rate and increases with time prolonged drug action, there is a time dependence. Reaches a certain inhibition rate (about 80%), the concentration of the platinum drug augmented Gallo, inhibition rate does not significantly increase, showing the platform effect. Of 3 lobaplatin cell cycle: same concentration lobaplatin role 24,48,72 hours S \u0026 G0/G1 phase compared with control group differences were statistically significant (p lt; 0.05). The 5μmol / L lobaplatin effect 24 hours began to cause laryngeal cancer cells in S phase was increased, along with the role of time to 72 hours when the decline in the proportion of cells in S phase, the increase in the proportion of G0/G1 phase cells. 20μmol / L lobaplatin role the 24 hours cells arrested in S phase blockade significantly longer duration of action, the decline in the proportion of S-phase cells, followed by a significant G0/G1 arrest, indicating that low concentrations of a short period of time laryngeal cancer cells arrested in S phase, high concentration for a long time laryngeal cancer cells can be arrested in G0/G1 phase. 4 lobaplatin apoptosis rate: normal control group cells apoptosis peak, 5 μmol / L role of the lobaplatin in small apoptotic peak after 24 hours, 48, 72 hours after the role appears obvious apoptosis peak , 20μmol / L for 24, 48 hours showed obvious apoptosis peak. The differences were statistically significant (p lt; 0.05) different the concentration lobaplatin role in laryngeal cancer cells apoptosis rates of 24, 48 and 72 hours same concentration lobaplatin, 24 hours of laryngeal cancer cells and apoptosis rate difference of 48 hours There are statistically significant (p lt; 0.05). Results table Mingluo platinum can induce apoptosis in laryngeal cancer cells, and apoptosis rate gradually increased with the increase in drug concentration and duration of action, of lobaplatin induced apoptosis in laryngeal cancer cells was time-and dose-dependent relationship, low dose lobaplatin ( 5μmol / L) 24 hours can induce apoptosis in laryngeal cancer cells. 5 Immunocytochemistry method (SP) to detect changes in groups of bcl-2 protein expression; 20 μmol / L the role of lobaplatin 48 hours after, bcl-2 expression weakened by the t-test p lt; 0.05, the difference was statistically significance. Conclusion: 1 lobaplatin, in vitro inhibition of laryngeal cancer cell proliferation in a time-and dose-dependent. 2 lobaplatin of laryngeal cancer cells produce cell cycle arrest, arrested in S phase, low concentrations of short high concentration for a long time can produce G0/G1 arrest. 3 lobaplatin laryngeal cancer cells can induce apoptosis in a time-and dose-dependent manner, 5μmol / the L concentration lobaplatin 24 hours laryngeal cancer cells can induce apoptosis. The 4 lobaplatin likely by down-regulating the expression of bcl-2, and induced apoptosis in laryngeal cancer cells which may be one of the mechanisms induced apoptosis lobaplatin.

The Initial Approach on Efect of Lobaplatin in Laryngeal Carcinoma Cell Line-Hep-2

Abstract

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