本文關(guān)鍵詞：細(xì)胞膜穿透肽TAT修飾抗癌肽HPRP-A1提高其抗癌選擇性的研究　出處：《吉林大學(xué)》2016年碩士論文　論文類型：學(xué)位論文

  更多相關(guān)文章： 抗癌肽 細(xì)胞穿透肽 TAT caspase依賴性 抗癌活性 治療指數(shù)

【摘要】：HPRP-A1是一種來自幽門螺旋桿菌核糖體蛋白L1(Rp L1)N端的α-螺旋抗癌肽。在本課題組前期的研究工作中已經(jīng)證實(shí),抗癌肽HPRP-A1既可以通過死亡受體介導(dǎo)的外源途徑誘導(dǎo)He La細(xì)胞凋亡,又可以通過線粒體介導(dǎo)的caspase依賴性內(nèi)源途徑誘導(dǎo)He La細(xì)胞凋亡。在本研究中,我們將細(xì)胞穿透肽TAT與抗癌肽HPRP-A1的C端進(jìn)行偶聯(lián)得到一種新的雜合肽,即HPRP-A1-TAT。與親本肽HPRP-A1相比,雜合多肽HPRP-A1-TAT對He La細(xì)胞表現(xiàn)出更高的抗癌活性,同時(shí)對人血紅細(xì)胞具有更低的溶血毒性,進(jìn)一步提高了其抗癌選擇性。同時(shí),我們利用化學(xué)合成方法將FITC標(biāo)記在多肽的氨基端(FITC-HPRP-A1和FITI-HPRP-A1-TAT),并通過激光共聚焦顯微鏡觀察和流式細(xì)胞儀檢測等方法對兩種多肽進(jìn)入細(xì)胞的過程和機(jī)制進(jìn)行探究。實(shí)驗(yàn)證實(shí),與親本肽HPRP-A1相比,雜合肽HPRP-A1-TAT可以更快速地通過胞吞和破膜兩種方式穿過細(xì)胞膜,進(jìn)入到細(xì)胞質(zhì)中。在同等濃度時(shí),HPRP-A1-TAT具有更高的抗癌活性,并且可以誘導(dǎo)更多的He La細(xì)胞產(chǎn)生早期凋亡,同時(shí)激活caspase蛋白酶活性。此外,多肽樣品加入He La細(xì)胞作用24h后,細(xì)胞中HPRP-A1-TAT的含量要明顯高于HPRP-A1。我們推測,可能是因?yàn)門AT具有更多陽離子氨基酸,對HPRP-A1有一定的保護(hù)作用,使其在細(xì)胞質(zhì)中降解緩慢,或是TAT-HPRP-A1可以通過胞吞形式進(jìn)入細(xì)胞內(nèi),從而使其對HPRP-A1具有一定的保護(hù)作用。本課題對HPRP-A1-TAT的抗癌作用機(jī)理進(jìn)行了初步探究,經(jīng)過TAT修飾的抗癌肽可以提高其活性,降低毒性,同時(shí)提高其治療指數(shù),為抗癌肽在臨床中的應(yīng)用提供了一種更好的策略和方向。
[Abstract]:HPRP-A1 is a kind of Helicobacter pylori ribosomal protein L1 (Rp L1) N terminal alpha helix anticancer peptides in this research. The group has confirmed that both HPRP-A1 anticancer peptides can induce apoptosis in He La cells by exogenous death receptor-mediated pathway, but also through the mitochondria mediated caspase dependent the endogenous pathway induced apoptosis of He La cells. In this study, we will cell penetrating peptide TAT and anticancer peptide HPRP-A1 C end was obtained by coupling a new hybrid peptide, compared to HPRP-A1-TAT. and peptide HPRP-A1, heterozygous He polypeptide HPRP-A1-TAT on La cells showed higher antitumor activity and hemolytic toxicity at the same time, the red blood cells in human blood is lower, to further enhance its antitumor selectivity. At the same time, we use the method of chemical synthesis of FITC markers in polypeptide amino terminal (FITC-HPRP-A1 and FITI-HPRP-A1-TAT), and by laser scanning Confocal microscopy observation and flow cytometry method to explore into the process and mechanism of cell of two polypeptide. Experiments show that compared with the parent peptide HPRP-A1, hybrid peptide HPRP-A1-TAT can more quickly across the cell membrane by endocytosis and membrane rupture in two ways, into the cytoplasm at the same concentration. When the HPRP-A1-TAT has higher antitumor activity and can induce more He La cells produced early apoptosis, and activation of caspase protease activity. In addition, adding He peptide samples La cells after 24h, HPRP-A1-TAT content in the cells to be significantly higher than the HPRP-A1., we speculate that may be because TAT has more cationic amino acids, protection effect on the HPRP-A1, the degradation in the cytoplasm is slow, or TAT-HPRP-A1 can enter cells via endocytosis, thus it has a protective effect on the HPRP-A1. The mechanism of anticancer action of HPRP-A1-TAT has been preliminarily studied. TAT modified anticancer peptide can improve its activity, reduce toxicity and improve its therapeutic index, providing a better strategy and direction for anticancer peptides in clinical application.

【學(xué)位授予單位】：吉林大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2016
【分類號】：R730.5

【參考文獻(xiàn)】

相關(guān)期刊論文 前2條

1 ZHAO LianJing;HUANG YiBing;GAO Song;CUI Yan;HE Dan;WANG Li;CHEN YuXin;;Comparison on effect of hydrophobicity on the antibacterial and antifungal activities of α-helical antimicrobial peptides[J];Science China(Chemistry);2013年09期

2 董兵;朱毅敏;;癌癥分子靶向治療的研究現(xiàn)狀[J];癌癥;2010年03期

，

本文編號：1361108