本文選題：蓮心堿　切入點：膀胱癌　出處：《桂林醫(yī)學(xué)院》2014年碩士論文　論文類型：學(xué)位論文

【摘要】：目的：通過蓮心堿（Liensinine）作用于膀胱癌細(xì)胞T24，，研究蓮心堿對膀胱癌細(xì)胞T24增殖及細(xì)胞周期的影響。同時初步探討蓮心堿調(diào)控膀胱癌細(xì)胞周期的機制，從而為用蓮心堿作為膀胱灌注輔助化療藥物的臨床應(yīng)用提供理論依據(jù)。方法：使用不同濃度的蓮心堿（1.5625、3.125、6.25、12.5、25μg/mL）處理膀胱癌細(xì)胞T24，通過CCK-8實驗及克隆形成實驗，檢測蓮心堿對細(xì)胞增殖的影響；對細(xì)胞進行PI染色后，然后使用流式細(xì)胞儀，檢測蓮心堿對T24細(xì)胞周期的影響；通過實時定量PCR技術(shù)及Western Blot技術(shù)檢測蓮心堿作用后T24細(xì)胞的p21基因的表達。結(jié)果：蓮心堿對膀胱癌細(xì)胞T24增殖有明顯的抑制作用：T24細(xì)胞抑制率分別為（-0.5±24.5）%、（26.51±14.28）%、（43.26±5）%、（42.93±8.51）%、（55.11±3.01）%。1.5625μg/mL蓮心堿組與同濃度甲醇對照組無明顯差異，無統(tǒng)計學(xué)意義（P＞0.05），3.125、6.25、12.5、25μg/mL蓮心堿組相對于甲醇對照組表現(xiàn)出明顯差異，且具有統(tǒng)計學(xué)意義（P0.05）；細(xì)胞周期檢測結(jié)果顯示，經(jīng)蓮心堿干預(yù)后，膀胱癌細(xì)胞T24的S期細(xì)胞群明顯增加，占有比例為（23.11±0.57）%、（24.91±1.10）%、（32.10±5.18）%、（53.09±3.91）%、（55.97±0.45）%，蓮心堿將T24細(xì)胞阻滯在S期；實時定量PCR檢測結(jié)果顯示，蓮心堿提高上調(diào)了T24細(xì)胞p21基因的表達水平。Westblot結(jié)果顯示：p21蛋白和磷酸化p53表達水平提高。結(jié)論：蓮心堿對膀胱癌細(xì)胞具有增殖抑制作用并將其阻滯在S期，該作用機制可能與p21及磷酸化p53表達上調(diào)有關(guān)。
[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 鹵24.55.51 鹵14.28 + 43.26 鹵53.26 鹵8.51L + 55.11 鹵3.01k.1.5625 渭 g/mL liensinine group, respectively.The inhibitory rate of liensinine on T24 cells was -0.5 鹵24.55.51 鹵14.28 鹵43.26 鹵53.26 鹵53.93 鹵8.511 鹵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 鹵0.57 + 24.91 鹵1.10, and 32.10 鹵5.18, 53.09 鹵3.91 and 55.97 鹵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.
【學(xué)位授予單位】：桂林醫(yī)學(xué)院
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2014
【分類號】：R737.14

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