菲律宾蛤仔寡肽抗前列腺癌DU-145细胞的机制研究
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浙江海洋大学,浙江海洋大学,浙江海洋大学,浙江海洋大学

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R282.77

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浙江省自然科学基金(LY12C20008)


Effect of oligopeptides from Ruditapes philippinarum on proliferation and apoptosis of human prostate cancer DU-145 cells
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#$TABFood Science and Pharmacy School of Zhejiang Ocean University,Zhejiang Provincial Key Engineering Technology Research Center of Marine Biomedical Products,#$TABFood Science and Pharmacy School of Zhejiang Ocean University,Zhejiang Provincial Key Engineering Technology Research Center of Marine Biomedical Products,#$TABFood Science and Pharmacy School of Zhejiang Ocean University,Zhejiang Provincial Key Engineering Technology Research Center of Marine Biomedical Products,#$TABFood Science and Pharmacy School of Zhejiang Ocean University,Zhejiang Provincial Key Engineering Technology Research Center of Marine Biomedical Products

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    摘要:

    为研究人工合成的菲律宾蛤仔寡肽体外抗前列腺癌细胞DU-145的活性及作用机制,实验采用流式细胞仪、激光共聚焦显微镜、Western blot等方法检测人工合成菲律宾蛤仔寡肽作用于DU-145细胞后细胞凋亡率、线粒体膜电位改变、细胞色素C表达、caspase3、caspase9的变化,并用电子显微镜观察细胞超微结构的改变。结果显示,随着药物浓度的增加和作用时间的延长,DU-145细胞的抑制率逐步增高,当药物浓度为2.5 mg/mL时,24 h抑制率为49.9%±4.1%,72 h抑制率为97.2%±7.34%;Hoechst33258染色和电子显微镜下观察发现,加药组细胞出现细胞核固缩、染色质凝集和细胞核碎片化等细胞凋亡的典型特征,随着药物浓度的增加,凋亡的细胞逐渐增多;线粒体膜电位逐步下降;Western blot检测显示药物作用后细胞内Bax表达增加,Bcl-2表达减少,Bax/Bcl-2比值增加,Cyt-C、caspase-3、caspase-9表达增加,且与药物浓度成正相关。研究表明,人工合成菲律宾蛤仔寡肽能诱导人前列腺癌细胞DU-145凋亡,可能机制为上调Bax表达、下调Bcl-2表达、增加Bax/Bcl-2比值,诱导线粒体膜电位下降,促使Cyt-C转移并激活caspase家族发生级联反应。

    Abstract:

    Thie paper aims to study the activity and mechanism of synthetic Ruditapes philippinarum oligopeptides(RPO) against prostate cancer DU-145 cells. After being incubated with RPO, DU-145 cells' apoptosis rate, changes in mitochondrial membrane potential, cytochrome C expression, changes of caspase3 and caspase9, cell ultrastructure changes were observed with flow cytometry, laser confocal microscope, Western blotting and electron microscope. The results showed that the factor to induce apoptosis of DU-145 cells is concentration and time dependent, when the RPO concentration was 2.5 mg/mL, the DU-145 cells inhibition rate was 49.9%±4.1% for 24 hours, 97.2%±7.3% for 72 hours. Those DU-145cells incubated with RPO were found to exhibit typical characteristics of apoptosis, such as nucleus psychosis, condensed chromatin, and nucleus fragmentation. The apoptosis rate of DU-145 cells rose as the concentration of RPO increased. Western blotting demonstrated that RPO increased the protein levels of Bax, caspase-3, caspase-9, Cyt-C and reduced the protein expression of Bcl-2. RPO can inhibit the proliferation and induce apoptosis of DU-145 cells possibly by up-regulating expression of Bax, down-regulating the expression of Bcl-2, reducing the Bcl-2/Bax ratio, inducing decline of mitochondrial membrane potential, promoting Cyt-C transfer and stimulating the caspase family cascade reaction.

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孙瑜,李连军,杨最素,丁国芳.菲律宾蛤仔寡肽抗前列腺癌DU-145细胞的机制研究[J].水产学报,2017,41(3):465~472

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  • 收稿日期:2016-09-10
  • 最后修改日期:2016-12-01
  • 录用日期:2017-03-15
  • 在线发布日期: 2017-03-17
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