莪术醇对膀胱癌细胞增殖和凋亡及EZH2表达的影响

发布时间：2018-05-14 22:30

本文选题：膀胱癌 + 莪术醇　；参考：《桂林医学院》2017年硕士论文

【摘要】：目的:我们前期研究证实EZH2基因在膀胱癌组织中高表达,在膀胱癌进展中有着重要的作用,而通过文献可知莪术醇能够对多种肿瘤的生长产生影响。因此通过体外实验研究莪术醇对两种人膀胱癌细胞系(EJ、T24)生长的影响,以及是否对这两种细胞系中EZH2的表达产生影响,初步探讨莪术醇在膀胱癌中可能的作用机制。方法:用不同浓度莪术醇(0、12.5、25、50、100mg/L)干预处理人膀胱癌EJ和T24细胞。1.使用CCK-8法检测莪术醇对EJ和T24细胞增殖抑制作用;2.通过克隆形成实验检测莪术醇对两种细胞克隆形成能力的影响;3.通过流式细胞术PI染色法测定莪术醇对EJ和T24细胞凋亡的影响;4.q RT-PCR法分析莪术醇对两种细胞中EZH2m RNA表达的影响;5.Western blot法检测两种细胞中EZH2蛋白的表达情况。结果:1.CCK-8法结果表明,莪术醇对两种膀胱癌细胞的增殖有明显的抑制作用,并且该作用随着药物浓度增加和孵育时间延长而增强。2.克隆形成实验结果表明,莪术醇处理组与对照组相比,细胞克隆形成能力下降,细胞增殖受到抑制。3.流式细胞术检测细胞凋亡率,结果显示膀胱癌EJ细胞凋亡率分别为:(0.50±0.05)%、(3.60±0.60)%、(6.20±1.00)%、(16.50±1.50)%、(29.00±2.15)%;T24细胞凋亡率为(2.40±0.10)%、(4.93±0.15)%、(6.03±0.15)%、(13.50±0.89)%、(25.80±0.66)%;莪术醇处理组EJ和T24细胞凋亡率与对照组相比均增加,并且随着莪术醇浓度增加其细胞凋亡率也逐渐增加,存在一定的浓度依赖性。4.q RT-PCR和Western blot法结果均显示莪术醇对膀胱癌EJ和T24细胞中EZH2基因的表达有明显抑制作用。结论:莪术醇对膀胱癌EJ和T24细胞有增殖抑制和诱导凋亡的作用,其机制可能与下调细胞中EZH2基因表达有关。
[Abstract]:Objective: our previous study confirmed the high expression of EZH2 gene in bladder cancer tissue, and it has an important role in the progression of bladder cancer. It is known that curcumaol can affect the growth of various tumors. Therefore, the effects of zedool on the growth of two human bladder cancer cell lines (EJ, T24) are studied in vitro. The expression of EZH2 in the two cell lines was affected, and the possible mechanism of curcumal in bladder cancer was preliminarily discussed. Methods: using different concentrations of zedoary alcohol (0,12.5,25,50100mg/L) to treat human bladder cancer EJ and T24 cells.1. using CCK-8 method to detect the proliferation inhibition of zedool to EJ and T24 cells, and 2. to detect zedoary by cloning and forming experiment. Effects of alcohol on the formation of two cell clones; 3. the effects of curcumal on the apoptosis of EJ and T24 cells were measured by flow cytometry PI staining; 4.q RT-PCR was used to analyze the effect of zedool on the expression of EZH2m RNA in two cells; 5.Western blot method was used to detect the expression of EZH2 protein in two cells. Results: 1.CCK-8 method results showed that zedoary The effect of alcohol on the proliferation of two bladder cancer cells was obviously inhibited, and the effect of this effect was enhanced with the increase of the concentration of drug and the prolongation of incubation time. The experimental results showed that the cell clone formation ability of zedorol treatment group was lower than that of the control group, and the cell proliferation was inhibited by the inhibition of.3. flow cytometry to detect the apoptosis rate of.2. cells. The apoptotic rate of EJ cells in bladder cancer cells was (0.50 + 0.05)%, (3.60 + 0.60)%, (6.20 + 1)%, (16.50 + 1.50)%, (29 + 2.15)%, T24 cell apoptosis rate was (2.40 + 0.10)%, (4.93 +%)%, (1%)%, (6.20)%, EJ and T24 cell apoptosis rate increased compared with the control group, and with zedoary zedoary zedoary The apoptosis rate of cells increased gradually. There was a certain concentration dependent.4.q RT-PCR and Western blot method. The results showed that curcumazol could inhibit the expression of EZH2 gene in EJ and T24 cells of bladder cancer. Conclusion: the effect of curcumal on the proliferation and apoptosis of EJ and T24 cells in bladder cancer is possible, and the mechanism may be possible. It is related to the downregulation of EZH2 gene expression in cells.

【学位授予单位】：桂林医学院
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R737.14

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