肿节风对人前列腺癌du145细胞pi3kaktmtor 信号传导通路的影响-effects of sarcandra glabra on pi 3 kaktmtor signaling pathway in human prostate cancer du 145 cells.docx

成都中医药大学博士学位论文肿节风对人前列腺癌细胞 成都中医药大学博士学位论文 肿节风对人前列腺癌细胞 PI3K/AKT/mTOR 信号通路的影响 PAGE PAGE 10 中文摘要 目 的：探讨中药肿节风对人雄激素非依赖性前列腺癌 DU-145 细胞增殖的影响以及对 DU-145 细胞增殖抑制的机理。 方 法：采用细胞培养技术，将肿节风胶囊配制成 2.5mg/ml、 1.25mg/ml 和 0.625mg/ml 三个不同浓度组的溶液，分别处理 DU-145 细胞 48 小时。以光镜观察 DU-145 细胞形态变化，以 MTT 法检测 DU-145 细胞增殖情况，以 ELLISA 法和 RT-PCR 法检测 PI3K、Akt、 mTOR 和 P70S6K 蛋白的浓度和表达情况。 结 果：肿节风溶液作用 DU-145 细胞 48 小时后，出现了细胞 凋亡的形态学改变，表现为数量减少、体积缩小、染色质凝集、核固 缩；DU-145 细胞增殖受到抑制，呈一定的量效关系；ELLISA 法检 测 DU-145 细胞 PI3K、Akt、mTOR 和 P70S6K 的吸光度 OD 值（代表 上述物质浓度）均不同程度降低；RT-PCR 法检测 DU-145 细胞 PI3K、 Akt、mTOR 和 P70S6K 的表达均不同程度降低。两种检测法得出结果 一致。 结 论：肿节风可破坏 DU-145 细胞正常生长，抑制 DU-145 细 胞增殖；其机制与其能降低促雄激素非依赖性前列腺癌发生与发展的 PI3K/Akt/mTOR 信号传导通路的关键因子 PI3K、Akt、mTOR 和 P70S6K 的浓度和表达有关。 关 键 词 ： 肿 节 风 ； 人 前 列 腺 癌 DU-145 细 胞 ； 细 胞 增 殖 PI3K/Akt/mTOR 信号传导通路；MTT；ELLISA；RT-PCR ABSTRACT Objectives：To observe the efficacy Sarcandra Glaba having on the multiplication of human DU-145 prostatic cancer cells in hormone- independent prostatic cancer as well as the principle of it restraining DU-145 cells to multiply. Methods：Cell cultivation was adopted with different groups using 2.5mg/ml, 1.25mg/ml and 0.625mg/ml Sarcandra Glaba concentrations on DU-145 cells for 48 hours each. The DU-145 cells’ morphology was observed under optic microscope. The multiplication of DU-145 cells was measured by MTT method. Both ELLISA and RT-PCR methods was used to measure the concentrations and expressions of PI3K, Akt, mTOR and P70S6K. Results：DU-145 cells’ morphologic changes were shown with number decreasing, volume reduction, chromatin coagulation and nucleus contraction after 48 hours’ influence Sarcandra Glaba concentrations had on them. A certain quantity-effect relation was shown in each concentration group. The values of OD of all PI3K, Akt, mTOR and P70S6K were lowed by the concentration groups measured with ELLISA. The expressions of all PI3K, Akt, mTOR and P70S6K were lowed by the concentration groups measured with RT-PCR. The two methods of