骨髓基质细胞HS-分泌物白细胞介素-对人急性髓细胞性白血病细胞系HL-0抗凋亡的作用

doi:10.16098/j.issn.0529-1356.2020.01.006

摘要/Abstract

摘要：

目的探讨骨髓基质细胞HS-分泌物白细胞介素-6(IL-6)急性髓细胞性白血病（AML）细胞系HL-0增殖与凋亡的影响。方法外培养HL-60细胞与HS-5细胞，构建共培养体系，应用扫描电子显微镜、ELISA法、细胞计数试剂盒8（CCK-8）法、AnnexinV-FITC/PI 双染流式细胞术、Real-time PCR和Western blotting技术分别检测HL-60细胞增殖和凋亡的变化；将不同组的HL-60细胞分别接种于BALB/c裸鼠，观察并记录成瘤情况。结果骨髓基质细胞HS-5分泌的IL-6能够促进HL-60细胞增殖、抑制其凋亡，并下调HL-60细胞中的促凋亡基因Bax的表达，上调抗凋亡基因Bcl-2的表达。共培养组HL-60细胞在BALB/c裸鼠中成瘤能力最强，HL-60细胞单独培养组成瘤能力最弱。结论骨髓基质细胞HS-5促进HL-60细胞增殖、抑制其凋亡的部分作用机制是通过分泌IL-6实现的。

关键词: 人急性髓细胞性白血病, 骨髓基质细胞, 白细胞介素-6, 免疫印迹法, 裸鼠

Abstract:

Objective To investigate the effect of interleukin(IL)-6 secreted by bone marrow stromal cells HS-5 on activity and apoptosis of human acute myeloid leukemia(AML) cells HL-60 and its possible mechanism. Methods HL-60 cells and HS-5 cells were cultured in vitro, and the co-culture system was established. Scanning electron microscope, ELISA, cell counting kit-8(CCK-8), AnnexinV-FITC/PI, double-staining flow cytometry, Real-time PCR and Western blotting techniques were used to detect the changes of viability and apoptosis of HL-60 cells, respectively. HL-60 cells from different groups were inoculated into BALB/c nude mice to observe and record tumorigenesis. Results IL-6 secreted by bone marrow stromal cells HS-5 could enhance the viability and inhibit the apoptosis of HL-60 cells, and down-regulate the expression of pro-apoptotic gene Bax and and up-regulate the expression of anti-apoptotic gene Bcl-2 in HL-60 cells. HL-60 cells in co-culture group had the strongest tumorigenicity in BALB/c nude mice, while HL-60 cells alone had the weakest tumorigenicity. Conclusion Part of the mechanism by which bone marrow stromal cells HS-5 promote the proliferation of HL-60 cells and inhibit their apoptosis may be through the secretion of IL-6.

Key words: Human acute myeloid leukemia, Bone marrow stromal cells, Interleukin-6, Western blotting, Nude mice

中图分类号:

R321.33

陈雅琳白天鸽李瑶瑶王文强魏虹. 骨髓基质细胞HS-分泌物白细胞介素-对人急性髓细胞性白血病细胞系HL-0抗凋亡的作用[J]. 解剖学报, 2020, 51(1): 32-39.

CHEN Ya-lin BAI Tian-ge LI Yao-yao WANG Wen-qiang WEI Hong. Anti-apoptotic effect of human acute myeloid leukemia cell line HL-60 stimulated by interleukin-6 from bone marrow stromal cells HS-5[J]. Acta Anatomica Sinica, 2020, 51(1): 32-39.

参考文献

［1］ Mambet C, Chivu-Economescu M, Matei L, et al. Murine models based on acute myeloid leukemia-initiating stem cells xenografting［J］. World J Stem Cells, 2018, 10(6): 57-65.

［2］ Saif A, Kazmi SFA, Naseem R, et al. Acute myeloid leukemia: is that all there is［J］? Cureus, 2018,10(8): e3198. 

［3］ Seckinger A, Hose D. Interaction between myeloma cells and bone tissue［J］. Radiologe, 2014,54(6): 545-550.

［4］ Xynos A, Corbella P, Belmonte N, et al. Bone marrow-derived hematopoietic cells undergo myogenic differentiation following a Pax-7 independent pathway［J］. Stem Cells, 2010,28(5): 965-973.

［5］ Wang JG, Chen XH, Zhou F, et al. Effect of main microenvironment components on chemosensitivity in bone marrow stromal cell-leukemia co-culture model［J］. Tissue Engineering Research and Clinical Rehabilitation in China, 2008(43): 8504-8508.(in Chinese)

王吉刚, 陈幸华, 周凡, 等.骨髓基质细胞白血病细胞共培养模型中主要微环境成分对化疗敏感性的影响［J］.中国组织工程研究与临床康复,2008(43): 8504-8508.

［6］ Ma QY, Gu P, Wang YY, et al. Protein mass spectrometry analysis of bone marrow stromal cell secretory protein［J］. Acta Anatomica Sinica, 2010,41(1):60-64. (in Chinese)

马芹颖,顾平,王彦永,等.骨髓基质细胞分泌蛋白的蛋白质质谱分析［J］.解剖学报,2010,41(1):60-64.

［7］ He GC, Deng R, Liu YL, et al. Serum levels of IFN-gamma, TGF-beta, IL-6 and IL-17 in patients with acute leukemia and their clinical significance［J］. Medical and Pharmaceutical Journal of Chinese People’s Liberation Army, 2017,29(6):19-21, 32. (in Chinese)

何光翠,邓锐,刘一岚,等.急性白血病患者血清IFN-γ、TGF-β、IL-6和IL-17水平及临床意义［J］.解放军医药杂志,2017,29(6):19-21, 32.

［8］ Hu XP, Liu XM. The role and clinical significance of serum IL-6, IL-8, IL-10 and TNF-alpha in the occurrence, metastasis and prognosis of breast cancer ［J］. Maternal and Child Health Care of China, 2018,33(23):5375-5378. (in Chinese)

胡香萍,刘贤明.血清IL-6、IL-8、IL-10与TNF-α在乳腺癌发生、转移及预后中的作用与临床意义［J］.中国妇幼保健,2018,33(23):5375-5378.

［9］ Chen P, Jin Q, Fu Q, et al. Induction of multidrug resistance of acute myeloid leukemia cells by cocultured stromal cells via upregulation of the PI3K/Akt signaling pathway［J］. Oncol Res, 2016, 24(4):215-223.

［10］Moshaver B, van der Pol MA, Westra AH, et al. Chemotherapeutic treatment of bone marrow stromal cells strongly affects their protective effect on acute myeloid leukemia cell survival ［J］. Leuk Lymphoma, 2008, 49(1): 134-148.

［11］Lü M, Zhou XQ, Lü YL, et al. Effect of IL-1 and IL-6 on adhesion of multiple myeloma cells to bone marrow stromal cells［J］. Shanghai Journal of Immunology, 2000(3):183. (in Chinese)

吕鸣, 周晓青, 吕有灵, 等. IL-1和IL-6对多发性骨髓瘤细胞与骨髓基质细胞粘附作用的影响［J］. 上海免疫学杂志, 2000(3):183.

［12］Frick JS, Zahir N, Muller M, et al. Colitogenic and non-colitogenic commensal bacteria differentially trigger DC maturation and Th cell polarization: an important role for IL6 ［J］. Eur J Immunol, 2006,36(6):15371547.

［13］Yang HX, Meng TT, Guan DF, et al. Effect of bone marrow stromal cell HS-5 on apoptosis of HL-60 cells ［J］. Journal of Shihezi University (Natural Science Edition),2015,33(5):588-592. (in Chinese)

杨红霞,孟腾腾,管东方,等.骨髓基质细胞HS-5对HL-60细胞凋亡的影响［J］.石河子大学学报(自然科学版),2015,33(05):588-592.

［14］Cui JW, Xi YZh. IL-6/IL-6R systemmediated targeted therapy strategy for leukemia ［J］. Chinese Journal of Experimental Hematology, 2001(2):184-187. (in Chinese)

崔建武,奚永志. IL-6/IL-6R系统介导靶向治疗白血病策略［J］. 中国实验血液学杂志, 2001(2):184-187.

［15］Kitajima S, Yoshida A, Kohno S, et al. The RB-IL-6 axis controls self-renewal and endocrine therapy resistance by fine-tuning mitochondrial activity［J］. Oncogene, 2017,36(36): 5145-5157.

［16］Gasche JA, Hoffmann J, Boland CR, et al. Interleukin-6 promotes tumorigenesis by altering DNA methylation in oral cancer cells［J］. Int J Cancer,2011,129(5): 10531063.

［17］Hu CL, Zhang ZhQ, Wen H, et al. Establishment of nude mice models of esophageal cancer cell lines with different metastatic potentials［J］. Gastroenterology, 2013, 18(9):548-551. (in Chinese)

胡翠梨,张志强,温浩,等. 不同转移潜能食管癌细胞株裸鼠模型的建立［J］.胃肠病学,2013,18(9):548-551.

［18］Xu YCh. Isolation, identification and regulation of hepatocellular carcinoma tumor stem cells ［D］. Chinese People’s Liberation Army Academy of Military Medical Sciences, 2011. (in Chinese)

许英晨. 肝癌肿瘤干细胞的分离鉴定与调控机制初探［D］.中国人民解放军军事医学科学院,2011.

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