CXCR2对胃癌细胞侵袭、迁移及凋亡的影响

摘要/Abstract

摘要：

目的：探究CXC类趋化因子受体2（C-X-C motif chemokine receptor, CXCR2）对胃癌细胞侵袭、迁移及凋亡等生物学特性的影响。方法：取人胃癌MGC80-3细胞和SGC-7901细胞。转染建立CXCR2过表达细胞,质粒瞬时转染和RNA干扰建立敲减细胞。蛋白质印迹实验研究蛋白质相关细胞表型变化。通过transwell侵袭及迁移实验、增殖实验（cell counting kit-8, CCK8）、流式细胞实验Annexin Ⅴ-PI双染法以及实时定量PCR等进行细胞功能、增殖、凋亡以及CXCR2 mRNA表达的检测。结果：胃癌细胞过表达CXCR2后其侵袭、迁移能力显著增强（P<0.05）,凋亡率显著降低（P<0.01）。CXCR2下调后抑制胃癌细胞的侵袭、迁移能力（P<0.05）,凋亡率显著升高（P=0.02）。CXCR2过表达的胃癌细胞中Akt的磷酸化水平显著上调。Akt磷酸化水平与凋亡抑制蛋白Bcl-2的表达水平相一致。结论：CXCR2上调后增强胃癌细胞的侵袭、迁移及抗凋亡能力。胃癌细胞中CXCR2的抗凋亡能力与CXCR2/Akt/Bcl-2通路相关。

关键词:CXC类趋化因子受体2,胃癌,侵袭,迁移,凋亡

Abstract:ObjectiveTo investigate the effects of C-X-C motif chemokine receptor 2 (CXCR2) on invasion, migration and apoptosis of gastric cancer cells.MethodsGastric cancer cells MGC80-3 and SGC-7901 with upregulated CXCR2 were established by stable Lentivirus transfection. CXCR2 gene silencing cells were established by plasmid transient transfection and RNA interference. Proteins related to cell phenotype changes were detected by Western blotting. The functional changes of cells were detected by transwell invasion and migration assays. Cell proliferation was measured by CCK8 assays. Apoptosis was detected by Annexin V-PI double staining of flow cytometry. CXCR2 mRNA was detected via real-time quantitative PCR.ResultsUpregulation of CXCR2 in gastric cancer cells exhibited an increased ability of cells migration and invasion (P<0.05) and reduced the apoptosis rate (P<0.01). Downregulation of CXCR2 inhibited the invasion and migration ability of gastric cancer cells(P<0.05) and increased the apoptosis rate(P=0.02). Phosphorylation level of Akt in gastric cancer cells which overexpressed CXCR2 was significantly upregulated. Phosphorylation level of Akt was consistent with the expression of apoptosis inhibitor protein Bcl-2.ConclusionsUpregulation of CXCR2 enhanced the ability of invasion, migration and evasion of apoptosis of gastric cancer cell. The evasion of apoptosis was associated with CXCR2/Aki/Bcl-2 pathway.

Key words:C-X-C motif chemokine receptor 2,Gastric cancer,Invasion,Migration,Apoptosis

中图分类号:

R735.2

陆伟辉, 刘威, 王聪, 王正林. CXCR2对胃癌细胞侵袭、迁移及凋亡的影响[J]. 外科理论与实践, 2021, 26(05): 430-436.

LU Weihui, LIU Wei, WANG Cong, WANG Zhenglin. Effects of CXCR2 on invasion, migration and apoptosis of gastric cancer cells[J]. Journal of Surgery Concepts & Practice, 2021, 26(05): 430-436.

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参考文献22

[1]	Rot A, von Andrian UH. Chemokines in innate and adaptive host defense: basic chemokinese grammar for immune cells[J]. Annu Rev Immunol, 2004, 22:891-928. doi:10.1146/annurev.immunol.22.012703.104543 URL
[2]	Cacalano G, Lee J, Kikly K, et al. Neutrophil and B cell expansion in mice that lack the murine IL-8 receptor homolog[J]. Science, 1994, 265(5172):682-684. pmid:8036519
[3]	Heidemann J, Ogawa H, Dwinell MB, et al. Angiogenic effects of interleukin 8 (CXCL8) in human intestinal microvascular endothelial cells are mediated by CXCR2[J]. J Biol Chem, 2003, 278(10):8508-8515. doi:10.1074/jbc.M208231200pmid:12496258
[4]	Robinson S, Cohen M, Prayson R, et al. Constitutive expression of growth-related oncogene and its receptor in oligodendrogliomas[J]. Neurosurgery, 2001, 48(4):864-873; pmid:11322447
	discussion 873-864. pmid:11322447
[5]	Bohrer LR, Schwertfeger KL. Macrophages promote fibroblast growth factor receptor-driven tumor cell migration and invasion in a CXCR2-dependent manner[J]. Mol Cancer Res, 2012, 10(10):1294-1305. doi:10.1158/1541-7786.MCR-12-0275pmid:22893608
[6]	Hiroaki K, Masakazu Y, Hirohisa N, et al. Clinicopathologic significance of the CXCL1-CXCR2 axis in the tumor microenvironment of gastric carcinoma[J]. PLoS One, 2017, 12(6):e0178635. doi:10.1371/journal.pone.0178635 URL
[7]	刘立, 盖金娜, 尹作文, 等. 趋化因子CXCL5调控NF-κB与Wnt/β-catenin信号通路抑制肿瘤免疫促进胃癌的机制研究[J]. 肿瘤防治研究, 2020, 47(5):340-345.
[8]	Wang Z, Liu H, Shen Z, et al. The prognostic value of CXC-chemokine receptor 2(CXCR2) in gastric cancer patients[J]. BMC Cancer, 2015, 15:766. doi:10.1186/s12885-015-1793-9 URL
[9]	Mantovani A, Allavena P, Sica A, et al. Cancer-related inflammation[J]. Nature, 2008, 454(7203):436-444. doi:10.1038/nature07205 URL
[10]	Mantovani A. The chemokine system: redundancy for robust outputs[J]. Immunol Today, 1999, 20(6):254-257. pmid:10354549
[11]	Allavena P, Germano G, Marchesi F, et al. Chemokines in cancer related inflammation[J]. Exp Cell Res, 2011, 317(5):664-673. doi:10.1016/j.yexcr.2010.11.013pmid:21134366
[12]	Lazennec G, Richmond A. Chemokines and chemokine receptors: new insights into cancer-related inflammation[J]. Trends Mol Med, 2010, 16(3):133-144. doi:10.1016/j.molmed.2010.01.003pmid:20163989
[13]	Waugh DJ, Wilson C. The interleukin-8 pathway in cancer[J]. Clin Can Res, 2008, 14(21):6735-6741. doi:10.1158/1078-0432.CCR-07-4843 URL
[14]	Wang RX, Ji P, Gong Y, et al. Value of CXCL8-CXCR1/2 axis in neoadjuvant chemotherapy for triple-negative breast cancer patients: a retrospective pilot study[J]. Breast Cancer Res Treat, 2020, 181(3):561-570. doi:10.1007/s10549-020-05660-z URL
[15]	Awaji M, Saxena S, Wu L, et al. CXCR2 signaling promotes secretory cancer-associated fibroblasts in pancrea-tic ductal adenocarcinoma[J]. FASEB J, 2020, 34(7):9405-9418. doi:10.1096/fj.201902990R URL
[16]	Murphy PM, Tiffany HL. Cloning of complementary DNA encoding a functional human interleukin-8 receptor[J]. Science, 1991, 253(5025):1280-1283. pmid:1891716
[17]	Holmes WE, Lee J, Kuang WJ, et al. Structure and functional expression of a human interleukin-8 receptor[J]. Science, 1991, 253(5025):1278-1280. pmid:1840701
[18]	Murphy PM, Baggiolini M, Charo IF, et al. International union of pharmacology.ⅩⅫ. Nomenclature for chemokine receptors[J]. Pharmacol Rev, 2000, 52(1):145-176. pmid:10699158
[19]	Murdoch C, Monk PN, Finn A. CXC chemokine receptor expression on human endothelial cells[J]. Cytokine, 1999, 11(9):704-712. pmid:10479407
[20]	Addison CL, Daniel TO, Burdick MD, et al. The CXC chemokine receptor 2, CXCR2, is the putative receptor for ELR+ CXC chemokine-induced angiogenic activity[J]. J Immunol, 2000, 165(9):5269-5277. doi:10.4049/jimmunol.165.9.5269pmid:11046061
[21]	徐佶, 张一楚. Bcl基因与Ki-67抗原在胃癌组织中表达的研究[J]. 外科理论与实践, 1998, 3(1):18-20.
[22]	Yang G, Rosen DG, Liu G, et al. CXCR2 promotes ova-rian cancer growth through dysregulated cell cycle, dimi-nished apoptosis, and enhanced angiogenesis[J]. Clin Can-cer Res, 2010, 16(15):3875-3886.