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。
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 measurement showed the same results.
Conclusions:Sarcandra Glaba concentrations can gave enough damage to the normal growth of DU-145 cells as well as restrain it to multiply. The principles of these effects can be related on the restraining effects of the concentrations and expressions of all PI3K, Akt, mTOR and P70S6K. The 4 factors above are regarded as the most important factors of PI3K/Akt/mTOR signal pathway which plays a critical role in the occurrence and development of hormone-independent prostatic cancer.
引文
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