基于差异蛋白质组学解析紫色杆菌素抑制结肠癌细胞HT29的作用机制
发布时间：2019-06-25 20:30
【摘要】：【目的】对紫色杆菌素作用后的结肠癌细胞HT29进行差异蛋白质组学分析,探究其影响的代谢通路,揭示其抑制癌细胞生长的作用机制,为新型抗癌药物的开发提供一定的参考。【方法】以不同剂量的紫色杆菌素处理HT29 24、48、72 h,通过MTT试验以及透射电镜分析其对结肠癌细胞的有效抑制剂量及抑制情况。在此基础上,对提取的3个处理组的蛋白进行同位素标记,利用反相液相色谱(RP-LC)联用串联质谱(AB SCIEX Triple TOF5600)获得样品中的多肽及其相对丰度信息,通过数据库(NCBI.Human.protein database)搜索比对,鉴定差异表达蛋白。利用GO分析、KEGG信号通路分析,解析紫色杆菌素抑癌作用代谢途径。【结果】紫色杆菌素对HT29的抑制作用呈一定的时间、剂量依赖性。当紫色杆菌素对HT29的抑制率达50%时,仅为阳性药物5-Fu剂量的1/6,具有更明显的抑制效果。电镜下观察显示,随着紫色杆菌素剂量增大,细胞内部线粒体出现空泡结构,质膜出泡;当浓度达30 mg·L~(-1)时,细胞膜消失,染色质边集。通过差异蛋白质组学分析,共鉴定出4 258个蛋白,表达差异在2倍以上的蛋白共为757个。其中高剂量组处理差异蛋白数为492个,低剂量组处理的差异蛋白数为112个,阳性对照组差异蛋白数为336个。KEGG分析显示这些差异蛋白共参与50条信号通路。其中有10条信号通路具有显著性(P0.05),主要参与核糖体循环途径、三羧酸循环途径和RNA降解途径等。【结论】紫色杆菌素主要通过影响HT29细胞生命活动中的蛋白转录和翻译水平进而发挥抑制作用。
[Abstract]:[objective] to analyze the differential proteome of colon cancer cell line HT29 treated with purple baculins, to explore the metabolic pathway of its effect, to reveal the mechanism of its inhibitory effect on the growth of cancer cells, and to provide some reference for the development of new anticancer drugs. [methods] the effective inhibitory dose and inhibition of HT29 on colon cancer cells were analyzed by MTT test and transmission electron microscope. [methods] HT29 was treated with different doses of purple bacteriocin for 48 h, 72 h, and its effective inhibitory dose and inhibition on colon cancer cells were analyzed by MTT test and transmission electron microscope. On this basis, the proteins of the three treatment groups were labeled by isotope, and the polypeptide and its relative abundance information were obtained by reversed-phase liquid chromatography (RP-LC) coupled with tandem mass spectrometry (AB SCIEX Triple TOF5600). The differentially expressed proteins were identified by database (NCBI.Human.protein database) search and comparison. GO analysis and KEGG signal pathway analysis were used to analyze the metabolic pathway of purple baculins in inhibiting cancer. [results] the inhibitory effect of purple baculins on HT29 was time-dependent and dose-dependent. When the inhibition rate of purple baculins on HT29 was 50%, it was only 1 鈮，
本文编号：2505981
[Abstract]:[objective] to analyze the differential proteome of colon cancer cell line HT29 treated with purple baculins, to explore the metabolic pathway of its effect, to reveal the mechanism of its inhibitory effect on the growth of cancer cells, and to provide some reference for the development of new anticancer drugs. [methods] the effective inhibitory dose and inhibition of HT29 on colon cancer cells were analyzed by MTT test and transmission electron microscope. [methods] HT29 was treated with different doses of purple bacteriocin for 48 h, 72 h, and its effective inhibitory dose and inhibition on colon cancer cells were analyzed by MTT test and transmission electron microscope. On this basis, the proteins of the three treatment groups were labeled by isotope, and the polypeptide and its relative abundance information were obtained by reversed-phase liquid chromatography (RP-LC) coupled with tandem mass spectrometry (AB SCIEX Triple TOF5600). The differentially expressed proteins were identified by database (NCBI.Human.protein database) search and comparison. GO analysis and KEGG signal pathway analysis were used to analyze the metabolic pathway of purple baculins in inhibiting cancer. [results] the inhibitory effect of purple baculins on HT29 was time-dependent and dose-dependent. When the inhibition rate of purple baculins on HT29 was 50%, it was only 1 鈮，
本文编号：2505981
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