KIF4A对巨噬细胞血管生成相关因子表达的调节作用

doi:10.6040/j.issn.1671-7554.0.2016.200

摘要/Abstract

摘要： 目的探讨驱动蛋白KIF4A对巨噬细胞不同亚群中血管生成相关因子表达的影响。方法 THP-1细胞在PMA及不同人重组细胞因子的外界刺激下分别分化为M0、M1、M2细胞,采用ELISA技术检测此3种巨噬细胞亚群中血管生成相关因子的表达水平。利用siRNA转染敲除M0、M1、M2细胞中KIF4A的表达,采用实时定量PCR检测敲除后血管生成相关因子的表达水平。结果巨噬细胞M0上清中可检测到LIF、VEGF、sVEGFR1及TIMP1,而sVEGFR2、Flt-3、Leptin、LeptinR、CD40L均不表达。其中LIF、VEGF、TIMP1在M0向M1、M2分化过程中,表达差异均无统计学意义(P>0.05);sVEGFR1在M0向M1分化过程中表达差异无统计学意义(P>0.05),向M2分化过程中表达显著上调(P<0.01)。转染KIF4A siRNA后,M0与M1细胞中LIF、sVEGFR1、VEGF、TIMP1的表达差异均无统计学意义(P>0.05);M2细胞中LIF、VEGF、TIMP1表达差异均无统计学意义(P>0.05),但sVEGFR1表达水平明显降低(P<0.05)。结论 M2细胞sVEGFR1表达水平显著上调;KIF4A参与M2细胞中sVEGFR1的表达。

关键词: 巨噬细胞, KIF4A, 转染, 血管生成相关因子

Abstract: Objective To explore the effect of KIF4A on expressions of angiogenesis-associated factors in M0, M1 and M2 cells. Methods THP-1 cells were differentiated into M0, M1 and M2 cells under the treatment of PMA and different recombination human cytokines in vitro. The expressions of several angiogenesis-associated factors in these three macrophage subpopulations were analyzed by ELISA. KIF4A siRNA was transfected into M0, M1 and M2 cells, and the levels of angiogenesis-associated factors after the transfection were analyzed by qRT-PCR. Results LIF, VEGF, sVEGFR1 and TIMP1 were detected from supernatant of M0 cells, whereas the expressions of sVEGFR2, Flt-3, Leptin, LeptinR and CD40L were not detected. The expressions of LIF, VEGF and TIMP1 did not significantly change during the differentiation from M0 to M1 or M2 cells(P>0.05). The expression of sVEGFR1 was also constant in M0 and M1 cells(P>0.05), but its expression was significantly up-regulated in M2 cells (P<0.01). After the transfection of KIF4A siRNA, the levels of LIF, sVEGFR1, VEGF and TIMP1 did not significantly change in M0 and M1 cells, and LIF, VEGF and TIMP1 in M2 cells(P>0.05). However, sVEGFR1 expression in M2 cells was significantly decreased after KIF4A siRNA transfection(P<0.05). Conclusion M2 cells exhibite higher sVEGFR1 expression and KIF4A is involved in sVEGFR1 expression in M2 cells.

Key words: Angiogenesis-associated factors, Macrophages, Transfection, KIF4A

中图分类号:

R392

陈意坤,王克涛,王华阳,邵倩倩,谈万业,宋晓彬,曲迅,魏奉才. KIF4A对巨噬细胞血管生成相关因子表达的调节作用[J]. 山东大学学报（医学版）, 2017, 55(1): 39-43.

CHEN Yikun, WANG Ketao, WANG Huayang, SHAO Qianqian, TAN Wanye, SONG Xiaobin, QU Xun, WEI Fengcai. Regulatory role of KIF4A in the expression of angiogenesis- associated factors of macrophages[J]. JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES), 2017, 55(1): 39-43.

参考文献

[1] Mantovani A, Sica A. Macrophages, innate immunity and cancer:balance, tolerance, and diversity[J]. Curr Opin Immunol, 2010, 22(2):231-237.
[2] Mills CD, Lenz LL, Harris RA. A breakthrough:macrophage-directed cancer immunotherapy[J]. Cancer Res, 2016, 76(3):513-516.
[3] Dirkx AE, Oude Egbrink MG, Wagstaff J, et al. Monocyte/macrophage infiltration in tumors:modulators of angiogenesis[J]. J Leukoc Biol, 2006, 80(6):1183-1196.
[4] Baay M, Brouwer A, Pauwels P, et al. Tumor cells and tumor-associated macrophages:secreted proteins as potential targets for therapy [J]. Clin Dev Immunol, 2011, 2011:565187.
[5] Bergers G, Hanahan D. Modes of resistance to anti-angiogenic therapy [J]. Nat Rev Cancer, 2008, 8(8):592-603.
[6] Ferrara N. Role of myeloid cells in vascular endothelial growth factor-independent tumor angiogenesis [J]. Curr Opin Hematol, 2010, 17(3):219-224.
[7] Verhey KJ,Hammond JW. Traffic control:regulation of kinesin motors [J]. Nat Rev Mol Cell Biol, 2009, 10(11):765-777.
[8] Yamada KH, Nakajima Y, Geyer M, et al. KIF13B regulates angiogenesis through Golgi to plasma membrane trafficking of VEGFR2 [J]. J Cell Sci, 2014, 127(Pt 20):4518-4530.
[9] Exertier P, Javerzat S, Wang B, et al. Impaired angiogenesis and tumor development by inhibition of the mitotic kinesin Eg5 [J]. Oncotarget, 2013, 4(12):2302-2316.
[10] Hanania R, Sun HS, Xu K, et al. Classically activated macrophages use stable microtubules for matrix metalloproteinase-9(MMP-9)secretion [J]. J Biol Chem, 2012, 287(11):8468-8483.
[11] Cornfine S, Himmel M, Kopp P, et al. The kinesin KIF9 and reggie/flotillin proteins regulate matrix degradation by macrophage podosomes [J]. Mol Biol Cell, 2011, 22(2):202-215.
[12] Wiesner C, Faix J, Himmel M, et al. KIF5B and KIF3A/KIF3B kinesins drive MT1-MMP surface exposure, CD44 shedding, and extracellular matrix degradation in primary macrophages[J]. Blood, 2010, 116(9):1559-1569.
[13] Zhao P, Gao D, Wang Q, et al. Response gene to complement 32(RGC-32)expression on M2-polarized and tumor-associated macrophages is M-CSF-dependent and enhanced by tumor-derived IL-4 [J]. Cell Mol Immunol, 2015, 12(6):692-699.
[14] Tahergorabi Z, Khazaei M. Leptin and its cardiovascular effects:focus on angiogenesis[J]. Adv Biomed Res, 2015, 4:79.
[15] Chakrabarti S, Rizvi M, Morin K, et al. The role of CD40L and VEGF in the modulation of angiogenesis and inflammation[J]. Vascul Pharmacol, 2010, 53(3-4):130-137.
[16] Ries C. Cytokine functions of TIMP-1 [J]. Cell Mol Life Sci, 2014, 71(4):659-672.
[17] 张静,刘晓惠. 可溶性血管内皮生长因子受体1 的研究进展 [J]. 中华临床医师杂志, 2015, 9(19):3625-3629.
[18] Ash J, McLeod DS, Lutty GA. Transgenic expression of leukemia inhibitory factor(LIF)blocks normal vascular development but not pathological neovascularization in the eye[J]. Mol Vis, 2005, 11:298-308.
[19] Melton DW, McManus LM, Gelfond JA, et al. Temporal phenotypic features distinguish polarized macrophages in vitro [J]. Autoimmunity, 2015, 48(3):161-176.
[20] Mazumdar M, Sundareshan S, Misteli T. Human chromokinesin KIF4A functions in chromosome condensation and segregation [J]. J Cell Biol, 2004, 166(5):613-620.
[21] Hu CK, Coughlin M, Field CM, et al. KIF4 regulates midzone length during cytokinesis[J]. Curr Biol, 2011, 21(10):815-824.
[22] Chen S, Han M, Chen W, et al. KIF1B promotes glioma migration and invasion via cell surface localization of MT1-MMP [J]. Oncol Rep, 2016, 35(2):971-977.
[23] Ghajar CM, George SC, Putnam AJ. Matrix metalloproteinase control of capillary morphogenesis[J]. Crit Rev Eukaryot Gene Expr, 2008, 18(3):251-278.
[24] Tabouret E, Boudouresque F, Farina P, et al. MMP2 and MMP9 as candidate biomarkers to monitor bevacizumab therapy in high-grade glioma [J]. Neuro Oncol, 2015, 17(8):1174-1176.
[25] Deryugina EI, Zajac E, Juncker-Jensen A, et al. Tissue-infiltrating neutrophils constitute the major in vivo source of angiogenesis-inducing MMP-9 in the tumor microenvironment[J]. Neoplasia, 2014, 16(10):771-788.

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