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HIFU通过下调miR-181a进而促进ICAM-1表达增强小鼠抗肿瘤免疫

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Keywords: 高强度聚焦超声,抗肿瘤免疫,microRNA,细胞间黏附分子-

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Abstract:

目的探讨miR-181a在高强度聚焦超声(high-intensityfocusedultrasound,HIFU)增强机体抗肿瘤免疫中的作用及其机制。方法①皮下构建供HIFU治疗的黑色素瘤移植瘤小鼠模型,采用简单随机法随机分为HIFU组与荷瘤组,每组30只,采用MTT方法检测各组脾淋巴细胞对B16细胞的杀伤活性,ELISA检测小鼠血清及共培养上清中TNF-α及IFN-γ的表达水平;②qRT-PCR检测HIFU组与荷瘤组脾淋巴细胞中的差异miRNAs;③用生物信息学方法预测差异miRNAs的潜在靶基因,通过RT-PCR及Westernblot验证靶基因的表达;④构建荧光素酶报告质粒进一步验证差异miRNAs的靶基因,再将差异miRNAs的mimics转染脾淋巴细胞,RT-PCR及Westernblot检测其靶基因表达的变化,以进一步确认差异miRNAs对靶基因表达的影响。结果①HIFU治疗能够抑制黑色素瘤生长,促进脾淋巴细胞分泌TNF-α和IFN-γ,增强脾淋巴细胞对肿瘤细胞的杀伤活性;②HIFU处理明显下调脾淋巴细胞中miR-181a的表达水平(P<0.01);③生物信息学方法预测发现共刺激分子细胞间黏附分子-1(intercellularadhesionmolecule-1,ICAM-1)可能是miR-181a的潜在靶基因,且HIFU处理明显促进脾淋巴细胞中ICAM-1的mRNA和蛋白质的表达(P<0.01);④miR-181amimics显著抑制pLUC-ICAM-1-3′UTR报告质粒荧光素酶的活性(P<0.01),并下调脾淋巴细胞中ICAM-1的mRNA(P<0.05)和蛋白质的表达(P<0.01)。结论HIFU通过抑制miR-181a对共刺激分子ICAM-1的负调控作用从而增强荷瘤小鼠的抗肿瘤免疫。

References

[1]  王智彪. 肿瘤治疗新方法-高强度聚焦超声肿瘤治疗技术[J]. 世界医疗器械,? 1995,? 5(11):? 35-38.? [2]Huang X,? Yuan F,? Liang M, et al. M-HIFU inhibits tumor growth,? suppresses STAT3 activity and enhances tumor specific immunity in a transplant tumor model of prostate cancer[J]. PLoS One,? 2012,? 7(7):? e41632.? [3]Xia J Z,? Xie F L,? Ran L F, et al. High-intensity focused ultrasound tumor ablation activates autologous tumor-specific cytotoxic T lymphocytes[J]. Ultrasound Med Biol,? 2012,? 38(8):? 1363-1371.? [4]Xu Z L,? Zhu X,? Lu P, et al. Activation of tumor-infiltrating antigen presenting cells by high intensity focused ultrasound ablation of human breast cancer[J]. Ultrasound Med Biol,? 2009,? 35(1): 50-57.? [5]Melero I,? Bach N,? Chen L. Costimulation,? tolerance and ignorance of cytolytic T lymphocytes in immune responses to tumor antigens[J]. Life Sci,? 1997,? 60(23):? 2035-2041.? [6]Jindra P T,? Bagley J,? Godwin J G, et al. Costimulation-dependent expression of microRNA-214 increases the ability of T cells to proliferate by targeting Pten[J]. J Immunol,? 2010,? 185(2):? 990-997.? [7]Parameswaran N,? Suresh R,? Bal V, et al. Lack of ICAM-1 on APCs during T cell priming leads to poor generation of central memory cells[J]. J Immunol,? 2005,? 175(4):? 2201-2211.? [8]Chen C Z,? Schaffert S,? Fragoso R, et al. Regulation of immune responses and tolerance:? the microRNA perspective[J]. Immunol Rev,? 2013,? 253(1):? 112-128.? [9]Vigorito E,? Kohlhaas S,? Lu D, et al. miR-155:? an ancient regulator of the immune system[J] . Immunol Rev,? 2013,? 253(1):? 146-157. [10]Podshivalova K, Salomon D R. MicroRNA regulation of T-lymphocyte immunity: modulation of molecular networks responsible for T-cell activation, differentiation, and development[J]. Crit Rev Immunol, 2013, 33(5): 435-476.? [11]Bronevetsky Y, Ansel K M. Regulation of miRNA biogenesis and turnover in the immune system[J]. Immunol Rev, 2013, 253(1): 304-316.? [12]周颦,? 苏立,? 傅敏,? 等. 高强度聚焦超声制备肿瘤疫苗对小鼠抗肿瘤免疫增强作用的研究[J]. 中国超声医学杂志,? 2012,? 28(7): 591-594.? [13]Gracias D T,? Stelekati E,? Hope J L, et al. The microRNA miR-155 controls CD8(+) T cell responses by regulating interferon signaling[J]. Nat Immunol,? 2013,? 14(6): 593-502.? [14]Qin A,? Wen Z,? Zhou Y, et al. MicroRNA-126 regulates the induction and function of CD4(+)Foxp3(+) regulatory T cells through PI3K/AKT pathway[J]. J Cell Mol Med,? 2013,? 17(2):? 252-264.? [15]Ueda R,? Kohanbash G,? Sasaki K, et al. Dicer-regulated microRNAs222 and 339 promote resistance of cancer cells to cytotoxic T-lymphocytes by down-regulation of ICAM-1[J]. Proc Natl Acad Sci USA,? 2009,? 106(26):? 10746-10751.? [16]den-Brok M H,? Sutmuller R P,? van-der-Voort R, et al. In situ tumor ablation creates an antigen source for the generation of antitumor immunity[J]. Cancer Res,? 2004,? 64(11):? 4024-4029.? [17]Wu F,? Wang Z B,? Lu P, et al. Activated anti-tumor immunity in cancer patients after high intensity focused ultrasound ablation[J]. Ultrasound Med Biol,? 2004,? 30 (9):? 1217-1222.? [18]Pietruczuk M,? Pietruczuk A,? Pancewicz S, et al. ICAM-1:? structure,? biological role and clinical significance[J]. Pol Merkur Lekarski,? 2004,? 17(101):? 507-511.? [19]van-de-Stolpe A,? van-der-Saag P T. Intercellular adhesion molecule-1[J]. J Mol Med (Berl),? 1996,? 74(1):? 13-33.? [20]Gonzalez P A,? Carreno L J,? Cespedes P F, et al. Modulation of tumor immunity by soluble and membrane-bound molecules at the immunological synapse[J]. Clin Dev Immunol,? 2013,? 2013:? 450291.
[2]  李静,沈宜,杨麟,等.小鼠肝癌细胞(H22)源外泌体的体内抗肝癌作用研究[J].第三军医大学学报,2008,30(19):1836.  LI Jing,SHEN Yi,YANG Lin,et al.Antitumor effects of exosomes derived from a mouse hepatoma carcinoma cell line H22[J].J Third Mil Med Univ,2008,30(01):1836.

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