EGCG对鼻咽癌CNE2细胞增殖抑制和凋亡诱导时MnSOD、NF-κB、Caspase3的表达变化

发布时间：2018-03-14 10:46

本文选题：表没食子儿茶素没食子酸酯　切入点：鼻咽肿瘤　出处：《桂林医学院》2010年硕士论文　论文类型：学位论文

【摘要】：目的初步探讨绿茶提取物EGCG对鼻咽癌CNE2细胞增殖和凋亡的影响,并分析该过程中MnSOD、NF-κB、Caspase-3的表达变化,为可能寻找一个有效的增敏药物与放疗配合使用来提高鼻咽癌的治疗效果。方法1.体外培养鼻咽癌低分化株CNE2,用不同质量浓度的(0、40、80、160mg/l)EGCG干预48h后,倒置显微镜下观察CNE2细胞的形态学改变。2.不同质量浓度的(0、40、80、160mg/l) EGCG作用CNE2细胞24、48、72h后,应用MTT法间接检测细胞增殖的变化,并计算抑制率。3.(0、40、80、160mg/l)EGCG作用CNE2细胞48h后,Hoechst33258染色观察凋亡细胞,并计算凋亡率;流式细胞仪检测EGCG对CNE2细胞周期的影响。4.RT-PCR检测EGCG抗鼻咽癌细胞株CNE2过程中NF-κB、MnSOD、Caspase-3的表达变化。结果1.EGCG处理CNE2细胞后,细胞分裂相变少,部分细胞变圆,随着浓度的增加,部分细胞出现漂浮及坏死的现象。EGCG能抑制鼻咽癌细胞CNE2的增殖,随着作用时间的延长和浓度的增加抑制作用越明显,表现为浓度与时间依赖性(P0.01)。2.EGCG能诱导细胞的凋亡,(0、40、80、160mg/l)EGCG处理CNE2细胞48h后,凋亡率分别为:(2.3±0.7)%、(13.7±1.1)%、(22.5±1.2)%、(36.2±2.1)%。与对照组比较,各实验组凋亡率明显增加(P0.01),各实验组两两比较差异显著(P0.01)。3.流式细胞术显示:(0、40、80、160mg/l)EGCG作用CNE2后,停滞在Go/G1期细胞百分率分别为:(53.2±1.4)%、(64.8±1.2)%、(71.9±1.0)%、(74.5±1.6)%。与对照组比较,各实验组G1细胞百分率明显增加(P0.01),各实验组两两比较差异显著(P0.01),呈浓度依赖性。4. (0、40、80、160mg/l)EGCG处理CNE2细胞48h后,NF-κBmRNA的表达呈下降趋势;MnSOD和Caspase-3mRNA的表达呈升高趋势。结论1.EGCG能抑制鼻咽癌细胞CNE2的增殖,并表现为浓度和时间依赖性;EGCG能诱导鼻咽癌CNE2的凋亡,将细胞阻滞于G0/G1,呈现为浓度依赖性。2.EGCG可能通过下调NF-κBmRNA和上调MnSOD和Caspase-3的表达来发挥其抗鼻咽癌的生物学活性。
[Abstract]:Objective to investigate the effect of green tea extract (EGCG) on the proliferation and apoptosis of nasopharyngeal carcinoma (NPC) CNE2 cells, and to analyze the expression of MnSODNF- 魏 BnCaspase-3 in nasopharyngeal carcinoma (NPC) cells. Methods 1. The poorly differentiated nasopharyngeal carcinoma cell line CNE2 was cultured in vitro. After 48 hours of treatment with different concentrations of NCNE, the morphologic changes of CNE2 cells were observed under inverted microscope. The CNE2 cells were treated with different concentrations of Rhizoma 4080,160 mg / L EGCG for 72 h. The proliferation of CNE2 cells was detected indirectly by MTT assay, and the inhibition rate of CNE2 cells was calculated by Hoechst33258 staining for 48 h after treatment with 40,80mg / L EGCG, and the apoptotic rate was calculated. Flow cytometry was used to detect the effect of EGCG on the cell cycle of CNE2. 4. RT-PCR was used to detect the expression of Caspase-3 in EGCG anti-nasopharyngeal carcinoma cell line CNE2. Results 1. After treated with EGCG, the proliferation of nasopharyngeal carcinoma (NPC) cells was inhibited by CNE2. 2. After treated with EGCG, the cell division and transformation were less and some cells became round. With the increase of concentration, some cells appeared floating and necrotic. EGCG could inhibit the proliferation of nasopharyngeal carcinoma cell line CNE2. The inhibitory effect of EGCG was more obvious with the prolongation of time and the increase of concentration. 2. EGCG could induce apoptosis of CNE2 cells in a dose-dependent and time-dependent manner. The apoptotic rates of CNE2 cells treated with EGCG for 48 h were 2.3 卤0.7 卤1.21 卤1.2 卤2.10.Compared with the control group, the apoptosis rates of the cells were significantly higher than those of the control group (P < 0.05), but not in the control group (P < 0.05), and compared with that in the control group (P < 0.05), the apoptotic rate was significantly higher than that in the control group (P < 0.05), and the apoptosis rate was significantly higher than that in the control group (P < 0.05). The apoptotic rate of each experimental group was significantly increased, and the difference was significant between two groups. Flow cytometry showed that the percentage of cells stagnant in the Go/G1 phase after treatment with CNE2 was 53.2 卤1.41 卤1.21.9 卤1.00.Compared with that of the control group, the percentage of cells in the Go/G1 phase was 74.5 卤1.60.Compared with the control group, the percentage of cells in the Go/G1 phase was 53.2 卤1.42 卤1.21.The percentage of the cells in the control group was 74.5 卤1.60.Compared with the control group, the percentage of cells in the Go/G1 phase was 53.2 卤1.42 卤1.21.9 卤1.0mg / L, respectively. The percentage of G1 cells in each experimental group increased significantly (P 0.01), and the difference between the two groups was significant in a concentration dependent manner. The expression of NF- 魏 BmRNA in CNE2 cells increased in a dose-dependent manner after 48 hours of treatment with 100mg / L EGCG. Conclusion 1. EGCG can inhibit the proliferation of nasopharyngeal carcinoma (NPC) cell line CNE2 in a concentration and time dependent manner. EGCG can induce the apoptosis of CNE2 in nasopharyngeal carcinoma. 2. Blocking cells at G _ 0 / G _ 1 in a concentration-dependent manner. 2. EGCG may exert its biological activity against nasopharyngeal carcinoma by down-regulating NF- 魏 BmRNA and up-regulating the expression of MnSOD and Caspase-3.
【学位授予单位】：桂林医学院
【学位级别】：硕士
【学位授予年份】：2010
【分类号】：R739.63

【参考文献】