KLF2調控S1P1～S1P5對類中性粒細胞遷移的作用機制_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; 類中性粒細胞; 遷移

DOI：

10.7507/1671-6205.202202009

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目的研究KLF2是否通過調控S1P1～S1P5影響類中性粒細胞遷移。方法構建攜帶有siRNA的慢病毒載體，特異性沉默KLF2基因，利用細胞遷移實驗檢測類中性粒細胞遷移情況，利用蛋白印跡法、實時PCR分別檢測KLF2、S1P1～S1P5蛋白和mRNA表達情況。結果 ① KLF2病毒干擾組較空白組、空載病毒組，遷移率明顯減少（P<0.05），S1P從0 μmol/L至0.1 μmol/L，隨著濃度的增加，各組類中性粒細胞遷移率都明顯增加（P<0.05），S1P從0.1 μmol/L至0.4 μmol/L ，隨著濃度的增加，各組類中性粒細胞遷移率基本相同（P>0.05）；② KLF2病毒干擾組與空白組、空載病毒組比較，S1P1蛋白及mRNA相對表達量減少（P<0.05）；③ KLF2病毒干擾組與空白組、空載病毒組比較，S1P2 、S1P3、S1P4、S1P5 mRNA表達量無差異（P>0.05）。結論 S1P對類中性粒細胞遷移的最佳濃度為0.1 μmol/L； KLF2通過正調控S1P1表達，促進類中性粒細胞遷移；在類中性粒細胞中KLF2不調控S1P2～S1P5表達。

Citation： 石丹丹, 朱黎明. KLF2調控S1P1～S1P5對類中性粒細胞遷移的作用機制. Chinese Journal of Respiratory and Critical Care Medicine, 2023, 22(2): 124-128. doi: 10.7507/1671-6205.202202009 Copy

1.	黃進, 朱黎明, 朱應群. KLF2/RelA比例失衡與哮喘患者中性粒細胞凋亡的關系研究. 中國呼吸與危重監護雜志, 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. 黃進, 朱黎明, 朱應群. KLF2/RelA比例失衡與哮喘患者中性粒細胞凋亡的關系研究. 中國呼吸與危重監護雜志, 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調控S1P1～S1P5對類中性粒細胞遷移的作用機制

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