KLF2调控S1P1～S1P5对类中性粒细胞迁移的作用机制_Chinese Journal of Respiratory and Critical Care Medicine

Authors：

 石丹丹 , 朱黎明

1. ;
2. ;

Corresponding author：

石丹丹, Email: 562452193@qq.com

Keywords：

KLF2; S1P1～S1P5; S1P; 类中性粒细胞; 迁移

DOI：

10.7507/1671-6205.202202009

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目的研究KLF2是否通过调控S1P1～S1P5影响类中性粒细胞迁移。方法构建携带有siRNA的慢病毒载体，特异性沉默KLF2基因，利用细胞迁移实验检测类中性粒细胞迁移情况，利用蛋白印迹法、实时PCR分别检测KLF2、S1P1～S1P5蛋白和mRNA表达情况。结果 ① KLF2病毒干扰组较空白组、空载病毒组，迁移率明显减少（P<0.05），S1P从0 µmol/L至0.1 µmol/L，随着浓度的增加，各组类中性粒细胞迁移率都明显增加（P<0.05），S1P从0.1 µmol/L至0.4 µmol/L ，随着浓度的增加，各组类中性粒细胞迁移率基本相同（P>0.05）；② KLF2病毒干扰组与空白组、空载病毒组比较，S1P1蛋白及mRNA相对表达量减少（P<0.05）；③ KLF2病毒干扰组与空白组、空载病毒组比较，S1P2 、S1P3、S1P4、S1P5 mRNA表达量无差异（P>0.05）。结论 S1P对类中性粒细胞迁移的最佳浓度为0.1 µmol/L； KLF2通过正调控S1P1表达，促进类中性粒细胞迁移；在类中性粒细胞中KLF2不调控S1P2～S1P5表达。

Citation： 石丹丹, 朱黎明. KLF2调控S1P1～S1P5对类中性粒细胞迁移的作用机制. Chinese Journal of Respiratory and Critical Care Medicine, 2023, 22(2): 124-128. doi: 10.7507/1671-6205.202202009 Copy

1.	黄进, 朱黎明, 朱应群. KLF2/RelA比例失衡与哮喘患者中性粒细胞凋亡的关系研究. 中国呼吸与危重监护杂志, 2018, 17(1): 1-5.
2.	Schwab SR, Cyster JG. Finding a way out: lymphocyte egress from lymphoid organs. Nat Immunol, 2007, 8(12): 1295-1301.
3.	Pappu R, Schwab SR, Cornelissen I, et al. Promotion of lymphocyte egress into blood and lymph by distinct sources of sphingosine-1-phosphate. Science, 2007, 316(5822): 295-298.
4.	Odumade OA, Weinreich MA, Jameson SC, et al. Krüppel-like factor 2 regulates trafficking and homeostasis of γδ T cells. J Immunol, 2010, 184(11): 6060-6066.
5.	Bai AL, Hu H, Yeung M, et al. Krüppel-like factor 2 controls T cell trafficking by activating L-selectin (CD62L) and sphingosine-1-phosphate receptor 1 transcription. J Immunol, 2007, 178(12): 7632-7639.
6.	Rahaman M, Costello RW, Belmonte KE, et al. Neutrophil sphingosine 1-phosphate and lysophosphatidic acid receptors in pneumonia. Am J Respir Cell Mol Biol, 2006, 34(2): 233-241.
7.	Buckley AF, Kuo CT, Leiden JM. Transcription factor LKLF is sufficient to program T cell quiescence via a c-Myc-dependent pathway. Nat Immunol, 2001, 2(8): 698-704.
8.	Fathalla AM, Chow SH, Naderer T, et al. Polymyxin-induced cell death of human macrophage-like THP-1 and neutrophil-like HL-60 cells associated with the activation of apoptotic pathways. Antimicrob Agents Chemother, 2020, 64(9): e00013-20.
9.	Shi PL, Li MH, Song C, et al. Neutrophil-like cell membrane-coated siRNA of lncRNA AABR07017145.1 therapy for cardiac hypertrophy via inhibiting ferroptosis of CMECs. Mol Ther Nucleic Acids, 2021, 27: 16-36.
10.	Zou WY, Chu XW, Cai CQ, et al. AKT-mediated regulation of polarization in differentiated human neutrophil-like HL-60 cells. Inflamm Res, 2012, 61(8): 853-862.
11.	Millius A, Weiner OD. Manipulation of neutrophil-like HL-60 cells for the study of directed cell migration. Methods Mol Biol, 2010, 591: 147-158.
12.	Richardson MW, Jadlowsky J, Didigu CA, et al. Kruppel-like factor 2 modulates CCR5 expression and susceptibility to HIV-1 infection. J Immunol, 2012, 189(8): 3815-3821.
13.	Lu YQ, Qin HX, Jiang B, et al. KLF2 inhibits cancer cell migration and invasion by regulating ferroptosis through GPX4 in clear cell renal cell carcinoma. Cancer Lett, 2021, 522: 1-13.
14.	Wani MA, Wert SE, Lingrel JB. Lung Kruppel-like factor, a zinc finger transcription factor, is essential for normal lung development. J Biol Chem, 1999, 274(30): 21180-21185.
15.	Santos-Beneit AM, Mollinedo F. Expression of genes involved in initiation, regulation, and execution of apoptosis in human neutrophils and during neutrophil differentiation of HL-60 cells. J Leukoc Biol, 2000, 67(5): 712-724.
16.	Hauert AB, Martinelli S, Marone C, et al. Differentiated HL-60 cells are a valid model system for the analysis of human neutrophil migration and chemotaxis. Int J Biochem Cell Biol, 2002, 34(7): 838-854.
17.	Hosoki K, Ying S, Corrigan C, et al. Analysis of a panel of 48 cytokines in BAL fluids specifically identifies IL-8 levels as the only cytokine that distinguishes controlled asthma from uncontrolled asthma, and correlates inversely with FEV₁. PLoS One, 2015, 10(5): e0126035.

1. 黄进, 朱黎明, 朱应群. KLF2/RelA比例失衡与哮喘患者中性粒细胞凋亡的关系研究. 中国呼吸与危重监护杂志, 2018, 17(1): 1-5.
2. Schwab SR, Cyster JG. Finding a way out: lymphocyte egress from lymphoid organs. Nat Immunol, 2007, 8(12): 1295-1301.
3. Pappu R, Schwab SR, Cornelissen I, et al. Promotion of lymphocyte egress into blood and lymph by distinct sources of sphingosine-1-phosphate. Science, 2007, 316(5822): 295-298.
4. Odumade OA, Weinreich MA, Jameson SC, et al. Krüppel-like factor 2 regulates trafficking and homeostasis of γδ T cells. J Immunol, 2010, 184(11): 6060-6066.
5. Bai AL, Hu H, Yeung M, et al. Krüppel-like factor 2 controls T cell trafficking by activating L-selectin (CD62L) and sphingosine-1-phosphate receptor 1 transcription. J Immunol, 2007, 178(12): 7632-7639.
6. Rahaman M, Costello RW, Belmonte KE, et al. Neutrophil sphingosine 1-phosphate and lysophosphatidic acid receptors in pneumonia. Am J Respir Cell Mol Biol, 2006, 34(2): 233-241.
7. Buckley AF, Kuo CT, Leiden JM. Transcription factor LKLF is sufficient to program T cell quiescence via a c-Myc-dependent pathway. Nat Immunol, 2001, 2(8): 698-704.
8. Fathalla AM, Chow SH, Naderer T, et al. Polymyxin-induced cell death of human macrophage-like THP-1 and neutrophil-like HL-60 cells associated with the activation of apoptotic pathways. Antimicrob Agents Chemother, 2020, 64(9): e00013-20.
9. Shi PL, Li MH, Song C, et al. Neutrophil-like cell membrane-coated siRNA of lncRNA AABR07017145.1 therapy for cardiac hypertrophy via inhibiting ferroptosis of CMECs. Mol Ther Nucleic Acids, 2021, 27: 16-36.
10. Zou WY, Chu XW, Cai CQ, et al. AKT-mediated regulation of polarization in differentiated human neutrophil-like HL-60 cells. Inflamm Res, 2012, 61(8): 853-862.
11. Millius A, Weiner OD. Manipulation of neutrophil-like HL-60 cells for the study of directed cell migration. Methods Mol Biol, 2010, 591: 147-158.
12. Richardson MW, Jadlowsky J, Didigu CA, et al. Kruppel-like factor 2 modulates CCR5 expression and susceptibility to HIV-1 infection. J Immunol, 2012, 189(8): 3815-3821.
13. Lu YQ, Qin HX, Jiang B, et al. KLF2 inhibits cancer cell migration and invasion by regulating ferroptosis through GPX4 in clear cell renal cell carcinoma. Cancer Lett, 2021, 522: 1-13.
14. Wani MA, Wert SE, Lingrel JB. Lung Kruppel-like factor, a zinc finger transcription factor, is essential for normal lung development. J Biol Chem, 1999, 274(30): 21180-21185.
15. Santos-Beneit AM, Mollinedo F. Expression of genes involved in initiation, regulation, and execution of apoptosis in human neutrophils and during neutrophil differentiation of HL-60 cells. J Leukoc Biol, 2000, 67(5): 712-724.
16. Hauert AB, Martinelli S, Marone C, et al. Differentiated HL-60 cells are a valid model system for the analysis of human neutrophil migration and chemotaxis. Int J Biochem Cell Biol, 2002, 34(7): 838-854.
17. Hosoki K, Ying S, Corrigan C, et al. Analysis of a panel of 48 cytokines in BAL fluids specifically identifies IL-8 levels as the only cytokine that distinguishes controlled asthma from uncontrolled asthma, and correlates inversely with FEV₁. PLoS One, 2015, 10(5): e0126035.

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KLF2调控S1P1～S1P5对类中性粒细胞迁移的作用机制

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