中药艾迪对人喉癌细胞Hep-2体外生长作用的研究

发布时间：2018-01-19 15:20

本文关键词： 中药艾迪人喉癌Hep-2细胞荧光定量PCR 环氧化酶-2　出处：《辽宁医学院》2012年硕士论文　论文类型：学位论文

【摘要】：目的研究中药艾迪对人喉癌Hep-2细胞的生长抑制作用及诱导凋亡作用方法本研究设实验组和对照组，实验组为0.4、0.8、1.0、2.0ug/ml的中药艾迪，对照组为未加药物即0ug/ml。实验组与对照组分别作用于人喉癌Hep-2细胞24h、48h及72h,四亚基噻唑蓝（MTT）法测定中药艾迪对人喉癌Hep-2细胞的生长抑制作用，通过Hoechst33258荧光染色后荧光显微镜下观察人喉癌细胞Hep-2细胞的凋亡状况，流式细胞仪（FCM）分析人喉癌Hep-2细胞的细胞周期及凋亡率的变化，荧光定量PCR法测定人喉癌Hep-2细胞中环氧化酶-2（COX-2）的表达量。结果 ①MTT检测示：实验组对人喉癌Hep-2细胞的生长有抑制作用，与对照组比较，差异具有统计学意义（P＜0.05），并有明显的时间和剂量依赖性。经0.4、0.8、1.0、2.0ug/ml作用72h后，其细胞抑制率分别为23.08%、37.42%、43.38%、54.48%。②荧光检测发现实验组可见典型的凋亡细胞形态特征，可见凋亡小体，正常细胞明显减少，，对照组未见明显细胞凋亡形态变化，可见正常细胞核为蓝色，染色质均匀分布。③流式细胞仪检测结果显示实验组作用72h后能增加G_0/G_1期的比例，减少S期和G_2/M的比例；实验组作用后72h后的细胞凋亡率依次上升，由28.62±1.23%增加到54.90±2.40%，呈剂量依赖性，差异具有统计学意义（P＜0.01）。④荧光定量PCR法检测，实验组人喉癌Hep-2细胞所表达的环氧化酶-2（COX-2）水平呈剂量递减关系。实验组中2.0ug/ml作用人喉癌Hep-2细胞后所表达的环氧化酶-2（COX-2）水平最低，分别为对照组倍数的0.950±0.07、0.847±0.05、0.369±0.02。与对照组比较差异有统计学意义（P＜0.05）。结论中药艾迪在体外能明显抑制人喉癌细胞Hep-2生长增殖及诱导其凋亡，诱导细胞凋亡的机制可能与通过下调COX-2的表达有关。
[Abstract]:Purpose Study on the effect of Aidi on the growth inhibition and apoptosis of Hep-2 cells of Human Laryngeal carcinoma Method In this study, the experimental group and the control group, the experimental group is 0.4m 0.8U 1.0g / ml of Chinese medicine Aidi. The experimental group and the control group were treated with Hep-2 cells for 24 h and 72 h, respectively. The inhibitory effect of Aidi on the growth of human laryngeal cancer Hep-2 cells was determined by tetra-subunit thiazolium blue TTassay. The apoptosis of human laryngeal carcinoma cell line Hep-2 was observed under fluorescence microscope after Hoechst33258 fluorescence staining. Flow cytometry (FCM) was used to analyze the changes of cell cycle and apoptosis rate in human laryngeal carcinoma Hep-2 cells. The expression of cyclooxygenase-2 and cyclooxygenase-2 in human laryngeal carcinoma Hep-2 cells was determined by fluorescence quantitative PCR. Results 1MTT assay showed that the growth of human laryngeal carcinoma Hep-2 cells was inhibited in the experimental group, and the difference was statistically significant compared with the control group (P < 0.05). The cell inhibition rates were 23.08% and 37.42%, respectively, after 72 hours of treatment with 0.48 渭 g / ml of 0.48 渭 g / ml and 1.0 渭 g / ml of 0.48 渭 g / ml for 72 h, and in a time-and dose-dependent manner, the inhibition rate of the cells was 23.08%. Fluorescence detection of 43.38 and 54.48.2 found typical apoptotic cell morphological features, apoptotic bodies, normal cells decreased significantly in the experimental group, but no obvious changes in apoptotic morphology were found in the control group. The results showed that the normal nucleus was blue and chromatin distributed evenly. 3. The results of flow cytometry showed that after 72 hours of treatment, the proportion of G _ (0) / G _ (1) phase was increased, and the ratio of S phase to G _ 2 / M was decreased in the experimental group. The apoptosis rate of the experimental group increased from 28.62 卤1.23% to 54.90 卤2.40 in a dose-dependent manner. The difference was statistically significant (P < 0.01). 4 fluorescence quantitative PCR method was used to detect. Expression of cyclooxygenase-2mCOX-2 in human laryngeal carcinoma Hep-2 cells in experimental group. The level of cyclooxygenase-2 (COX-2) was the lowest in the experimental group (2.0ugml / ml) after the treatment of human laryngeal carcinoma Hep-2 cells. The ratio of the control group was 0.950 卤0.07, 0.847 卤0.05, 0.369 卤0.02.Compared with the control group, the difference was statistically significant (P < 0.05). Conclusion Aidi can obviously inhibit the growth and proliferation of human laryngeal carcinoma cell line Hep-2 and induce its apoptosis in vitro. The mechanism of inducing apoptosis may be related to the down-regulation of COX-2 expression.
【学位授予单位】：辽宁医学院
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R739.65

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