莲心碱对膀胱癌细胞增殖的影响

发布时间：2018-03-22 18:46

本文选题：莲心碱　切入点：膀胱癌　出处：《桂林医学院》2014年硕士论文　论文类型：学位论文

【摘要】：目的：通过莲心碱（Liensinine）作用于膀胱癌细胞T24，，研究莲心碱对膀胱癌细胞T24增殖及细胞周期的影响。同时初步探讨莲心碱调控膀胱癌细胞周期的机制，从而为用莲心碱作为膀胱灌注辅助化疗药物的临床应用提供理论依据。方法：使用不同浓度的莲心碱（1.5625、3.125、6.25、12.5、25μg/mL）处理膀胱癌细胞T24，通过CCK-8实验及克隆形成实验，检测莲心碱对细胞增殖的影响；对细胞进行PI染色后，然后使用流式细胞仪，检测莲心碱对T24细胞周期的影响；通过实时定量PCR技术及Western Blot技术检测莲心碱作用后T24细胞的p21基因的表达。结果：莲心碱对膀胱癌细胞T24增殖有明显的抑制作用：T24细胞抑制率分别为（-0.5±24.5）%、（26.51±14.28）%、（43.26±5）%、（42.93±8.51）%、（55.11±3.01）%。1.5625μg/mL莲心碱组与同浓度甲醇对照组无明显差异，无统计学意义（P＞0.05），3.125、6.25、12.5、25μg/mL莲心碱组相对于甲醇对照组表现出明显差异，且具有统计学意义（P0.05）；细胞周期检测结果显示，经莲心碱干预后，膀胱癌细胞T24的S期细胞群明显增加，占有比例为（23.11±0.57）%、（24.91±1.10）%、（32.10±5.18）%、（53.09±3.91）%、（55.97±0.45）%，莲心碱将T24细胞阻滞在S期；实时定量PCR检测结果显示，莲心碱提高上调了T24细胞p21基因的表达水平。Westblot结果显示：p21蛋白和磷酸化p53表达水平提高。结论：莲心碱对膀胱癌细胞具有增殖抑制作用并将其阻滞在S期，该作用机制可能与p21及磷酸化p53表达上调有关。
[Abstract]:Aim: to study the effects of liensinine on the proliferation and cell cycle of bladder cancer cell line T24, and to explore the mechanism of Liensinine in regulating the cell cycle of bladder cancer. So as to provide the theoretical basis for the clinical application of liensinine as an adjuvant chemotherapeutic drug for bladder instillation. Methods: T24 cells were treated with different concentrations of liensinine at 1.5625 ~ 3.125 ~ 6.125 渭 g 路mL ~ (-1) and 25 渭 g 路mL ~ (-1), respectively. T24 cells were treated by CCK-8 assay and clone formation assay. After Pi staining, flow cytometry was used to detect the effect of liensinine on the cell cycle of T24 cells. The expression of p21 gene in T24 cells was detected by real-time quantitative PCR and Western Blot. Results: the inhibitory rate of liensinine on T24 proliferation of bladder cancer cell line was -0.5 卤24.55.51 卤14.28 + 43.26 卤53.26 卤8.51L + 55.11 卤3.01k.1.5625 渭 g/mL liensinine group, respectively.The inhibitory rate of liensinine on T24 cells was -0.5 卤24.55.51 卤14.28 卤43.26 卤53.26 卤53.93 卤8.511 卤3.01k.1.5625 渭 g/mL, respectively. There was no significant difference between the same concentration of methanol and the control group. There was no significant difference between the Liensinine group and the methanol control group (P > 0.05), and the cell cycle test showed that the S phase cell population of bladder cancer cell line T24 increased obviously after the intervention of Liensinine, and there was a significant difference between the Lianxin group and the methanol control group (P < 0.05), and the results of cell cycle test showed that the S phase of T24 cells was significantly increased after the intervention of Liensinine. The percentage of T24 cells was 23.11 卤0.57 + 24.91 卤1.10, and 32.10 卤5.18, 53.09 卤3.91 and 55.97 卤0.45%, respectively, and T24 cells were blocked in S phase by liensinine, and the results of real-time quantitative PCR showed that T24 cells were in S phase. Liensinine upregulated the expression of p21 gene in T24 cells. Western blot showed that the expression of p21 protein and phosphorylated p53 were increased in T24 cells. Conclusion: liensinine can inhibit the proliferation of bladder cancer cells and block it in S phase. This mechanism may be related to the up-regulation of p21 and phosphorylated p53 expression.
【学位授予单位】：桂林医学院
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R737.14

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