發(fā)布時(shí)間：2018-04-25 19:17

  本文選題：胸腺肽α + 急性髓系白血病　； 參考：《中華腫瘤防治雜志》2017年02期

【摘要】：目的吲哚胺2,3-雙加氧酶(indoleamine 2,3-dioxygenase,IDO)是色氨酸代謝過(guò)程中的限速酶,近年來(lái)研究發(fā)現(xiàn),IDO高表達(dá)是急性髓系白血病(acute myeloid leukemia,AML)細(xì)胞介導(dǎo)免疫耐受的重要機(jī)制,因此研究如何調(diào)控IDO表達(dá),將有可能為AML治療提供新的輔助方法。本實(shí)驗(yàn)擬從分子生物及蛋白質(zhì)水平,探討胸腺肽α1(thymosin-α1,Tα1)調(diào)節(jié)AML細(xì)胞株IDO表達(dá)及改善T細(xì)胞的免疫抑制作用,為AML免疫治療提供理論依據(jù)。方法采用逆轉(zhuǎn)錄-聚合酶聯(lián)反應(yīng)(RT-PCR)檢測(cè)γ-干擾素(interferon-γ,IFN-γ)、Tα1對(duì)HL-60/K562細(xì)胞株IDO mRNA表達(dá)的影響;采用蛋白質(zhì)印跡法檢測(cè)IFN-γ和Tα1對(duì)HL-60/K562細(xì)胞株IDO蛋白表達(dá)的影響;采用MTT檢測(cè)Tα1和1-甲基色氨酸(1-methyl-tryptophan,1-MT)作用后經(jīng)IFN-γ誘導(dǎo)的HL-60/K562細(xì)胞對(duì)植物血凝素(phytohaemagglutinin,PHA)刺激T細(xì)胞增殖抑制作用。結(jié)果HL-60/K562細(xì)胞株均表達(dá)IDO mRNA;IFN-γ作用后,IDO mRNA表達(dá)明顯增加(HL-60細(xì)胞株t=130.19,K562細(xì)胞株t=67.38,均P0.05);IFN-γ+Tα1共同作用后,明顯下調(diào)IDO mRNA表達(dá)(HL-60細(xì)胞株F=132.55,K562細(xì)胞株F=77.36,均P0.05),并呈濃度依賴性(HL-60細(xì)胞株r=-0.71,K562細(xì)胞株r=-0.80);HL-60/K562細(xì)胞株低水平表達(dá)IDO蛋白;經(jīng)IFN-γ作用后,HL-60細(xì)胞株IDO蛋白表達(dá)水平為2.505 6±0.019 6,明顯高于空白對(duì)照組0.189 7±0.083 4,t=326.79,P0.05;K562細(xì)胞株IDO蛋白表達(dá)水平為0.988 8±0.023 9,明顯高于空白對(duì)照組0.253 6±0.016 3,t=76.12,P0.05;IFN-γ+Tα1共同作用后,經(jīng)IFN-γ+Tα1共同作用后,IDO蛋白在HL-60細(xì)胞株各組的表達(dá)水平分別為2.455 0±0.066 1、1.526 0±0.017 3、1.134 1±0.060 1、0.806 0±0.029 7和0.070 0±0.003 6,并呈濃度依賴性,F=4 187.76,r=-0.77,P0.05;經(jīng)IFN-γ+Tα1共同作用后,IDO蛋白在K562細(xì)胞株各組的表達(dá)水平分別為0.998 6±0.016 4、0.784 3±0.014 9、0.714 5±0.009 5、0.656 6±0.007 6和0.492 2±0.010 9,并呈濃度依賴性,F=59.50,r=-0.76,P0.05。IFN-γ誘導(dǎo)的HL-60/K562細(xì)胞可抑制PHA刺激的T細(xì)胞增殖(P0.05),并呈濃度依賴性(HL-60細(xì)胞株r=0.99,K562細(xì)胞株r=0.98);1-MT組和Tα1組中HL-60/K562細(xì)胞株對(duì)T細(xì)胞增殖的抑制率明顯低于對(duì)照組,差異有統(tǒng)計(jì)學(xué)意義,P0.05。結(jié)論 IDO可能在AML誘導(dǎo)機(jī)體產(chǎn)生免疫耐受過(guò)程中起重要作用,Tα1可下調(diào)AML細(xì)胞IDO表達(dá),改善AML細(xì)胞對(duì)T細(xì)胞的免疫抑制作用。
[Abstract]:Objective Indoleamine 2ndoleamine 23-dioxygenase (IDO) is a rate-limiting enzyme in tryptophan metabolism. In recent years, it has been found that the high expression of indoleamine-3-dioxygenase (IDO) is an important mechanism of immune tolerance mediated by acute myeloid leukemiaMLs in acute myeloid leukemia (AML) cells, so we studied how to regulate the expression of IDO. It will be possible to provide new adjuvant methods for AML therapy. The aim of this study was to investigate the effects of thymosin- 偽 1T 偽 1) on regulating the expression of IDO in AML cell line and improving the immunosuppressive effect of T cell line from the molecular biological and protein levels, and to provide a theoretical basis for the immunotherapy of AML. Methods reverse transcription-polymerase chain reaction (RT-PCR) was used to detect the effect of interferon- 緯 -interferon- 緯 on the expression of IDO mRNA in HL-60/K562 cell line, and the effect of IFN- 緯 and T 偽 1 on the expression of IDO protein in HL-60/K562 cell line was detected by Western blotting. MTT was used to detect the inhibitory effect of HL-60/K562 cells induced by IFN- 緯 on the proliferation of T cells stimulated by phytohaemagglutinin (PHAs). Results all HL-60/K562 cell lines expressed IDO mRNA-IFN- 緯. After treatment, the expression of tndo mRNA was significantly increased in HL-60 cell line (tn130.19) K562 cell line tnr 67.38, and both P0.05 and IFN- 緯 T 偽 1. The expression of IDO mRNA in HL-60 cell line was significantly down-regulated, and the expression of IDO protein in HL-60 cell line was significantly lower than that in HL-60 cell line (P 0.05), and the expression of IDO protein in HL-60 / K562 cell line was significantly lower than that in HL-60 / K562 cell line (P < 0.05). The expression of IDO protein in HL-60 / K562 cell line was significantly lower than that in HL-60 / K562 cell line in a dose-dependent manner. The expression level of IDO protein in HL-60 cell line was 2.505 6 鹵0.019 6 after treatment with IFN- 緯, which was significantly higher than that in the control group (0.189 7 鹵0.083 4 TX 326.79) P0.05 K562 cell line (0.988 8 鹵0.023 9), which was significantly higher than that in the blank control group (0.253 6 鹵0.016 3t76.12P0.05) IFN- 緯 T 偽 1, and the expression of IDO protein in HL-60 cell line was significantly higher than that in control group (0.988 8 鹵0.023 9). After co-treatment with IFN- 緯 T 偽 1, the expression levels of ido protein in each HL-60 cell line were 2.455 鹵0.066 1a 1.526 鹵0.017 31.134 1 鹵0.060 1U 0.8060 鹵0.029 7 and 0.070 0 鹵0.003 6, respectively, in a concentration-dependent manner, and the expression of Ido protein in K562 cell line was observed in a dose-dependent manner after co-treatment of IFN- 緯 T 偽 1 and IFN- 緯 T 偽 1 in K562 cell line. In a dose-dependent manner, HL-60/K562 cells induced by FN 59.50r-0.76N P0.05.IFN- 緯 could inhibit the proliferation of PHA stimulated T cells (P0.055.05), and in a concentration-dependent manner, the HL-60/K562 cells in the HL-60/K562 group and T 偽 1 group could inhibit the proliferation of T cells induced by PHA, and in a concentration-dependent manner, the proliferation of HL-60/K562 cells in the HL-60 cell line r0.981-MT and T 偽 1 group was inhibited by P0. 055. 0. 0. 7. 843 鹵0.014 9, 0. 784 3 鹵0.014 9, 0. 714 5 鹵0.009 5. 6 鹵0.007 6 鹵0.007 6 6 and 0.492 2 鹵0.010 9, respectively. The inhibitory rate of T cell proliferation was significantly lower than that of control group. The difference was statistically significant (P 0.05). Conclusion IDO may play an important role in the induction of immune tolerance by AML. T 偽 1 can down-regulate the expression of IDO in AML cells and improve the immunosuppressive effect of AML cells on T cells.
【作者單位】： 四川省醫(yī)學(xué)科學(xué)院·四川省人民醫(yī)院兒科;四川省腫瘤醫(yī)院腫瘤研究所;
【基金】：四川省衛(wèi)生廳科研基金(090424)
【分類號(hào)】：R733.71

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