EphA1/EphrinA反馈环调控内皮祖细胞促肝细胞癌血管生成潜能

doi:10.3969/j.issn.2095-9400.2019.11.002

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Abstract Objective: To study the effect of regulated EphA1/EphrinA1 signaling pathways in endothelial progenitor cells (EPCs) on their cell biological behavior and promotion of angiogenesis potency in hepatocellular carcinoma. Methods: Different concentrations of doxycycline were used to regulate the EphA1 gene express in EPCs line EPCsEphA1/SiRNA-Tet in vitro and in vivo. Western blot was used to test the EphA1/EphrinA1 signaling pathway variation; CCK8 method, wound healing and cell invasion assay were used to test the changes in cell proliferation, migration and invasion ability; xenograft tumor growth curve and immunohistochemical staining of tumor microvascular were used to test their promote angiogenesis potency in hepatocellular carcinoma. T test was used to compare parameters between the groups. Results: In vitro assay it was showed that 10 μg/mL doxycycline could inhibit EphA1/EphrinA1 signaling pathway activity to the maximum extent. Compared with β-actin, the gray value of EphA1 and its ligand EphrinA1 protein expressions were 0.293±0.029 and 0.603±0.038 respectively, being statistically different (t=12.940, 4.864; P<0.001, 0.008) from Dox(-) group 0.943±0.041 and 0.960±0.062. The results of cell biological behavior showed that regulated EphA1 gene expression in EPCs had no effected on their proliferation ability. Compared with Dox(-) group, 10 μg/mL doxycycline regulation could inhibit EPCs’ motility and invasion ability (t=4.435, 2.467; P=0.002, 0.039) to the maximum extent. 100 μg/mL doxycycline induction in vitro could decrease to capacity EphA1 gene expression in EPCs in vivo. Compared with tumor volume (924.5±81.8) mm3 in Dox(-) group, the regulated group tumor volume (543.8±24.6) mm3 of nude mice was decreased at the end of sixth week (t=4.909, P=0.001) and compared with Dox(-) group (37.0±4.1), the microvascular density in regulated group 21.6±3.6 was significantly decreased (t=2.823, P=0.024). Conclusion: Different concentrations of doxycycline can precisely regulate the activity of EphA1/EphrinA1 signaling axis in EPCs in vitro and in vivo, thus controlling the promoted angiogenesis potency in hepatocellular carcinoma.

Key words： hepatocellular carcinoma endothelial progenitor cells angiogenesis EphA1/EphrinA1

Received: 25 June 2019

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	YU Haitao
	GUO Pengyi
	XIE Xiaozai
	CHEN Gang

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https://xb.wmu.edu.cn/EN/10.3969/j.issn.2095-9400.2019.11.002 OR https://xb.wmu.edu.cn/EN/Y2019/V49/I11/791

[1] 陈钢, 金炜东, 王怡, 等. EphA1基因可控性双稳转内皮祖细胞系EPCsTet-On-EphA1SiRNA的建立[J]. 肝胆胰外科杂志, 2012, 24(3): 220-223.
[2] DEBATIN K M, WEI J, BELTINGER C. Endothelial progenitor cells for cancer gene therapy[J]. Gene Ther, 2008, 15(10): 780-786.
[3] OH H K, HA J M, O E, et al. Tumor angiogenesis promoted by ex vivo differentiated endothelial progenitor cells is effectively inhibited by an angiogenesis inhibitor, TK1-2[J]. Cancer Res, 2007, 67(10): 4851-4859.
[4] GAO D, NOLAN D J, MELLICK A S, et al. Endothelial progenitor cells control the angiogenic switch in mouse lung metastasis[J]. Science, 2008, 319(5860): 195-198.
[5] WEI J, BLUM S, UNGER M, et al. Embryonic endothelial progenitor cells armed with a suicide gene target hypoxic lung metastases after in avenous delivery[J]. Cancer Cell, 2004, 5(5): 477-488.
[6] MELLICK A S, PLUMMER P N, NOLAN D J, et al. Using the transcription factor inhibitor of DNA binding 1 to selectively target endothelial progenitor cells offers novel strategies to inhibit tumor angiogenesis and growth[J]. Cancer Res, 2010, 70(18): 7273-7282.
[7] LEE R J, SPRINGER M L, BLANCO-BOSE W E, et al. VEGF gene delivery to myocardium:deleterious effects of unregulated expression[J]. Circulation, 2000, 102(8): 898-901.
[8] GOVERDHANA S, PUNTEL M, XIONG W, et al. Regulatable gene expression systems for gene therapy applications: progress and future challenges[J]. Mol Ther, 2005, 12(2): 189-211.
[9] BARDE I, ZANTA-BOUSSIF M A, PAISANT S, et al. Efficient control of gene expression in the hematopoietic system using a single Tet-on inducible lentiviral vector[J]. Mol Ther, 2006, 13(2): 382-390.
[10] 陈钢, 王怡, 易继林, 等. 肝细胞癌中Ephrin-A1及其受体的表达与血管生成的关系[J]. 中国普通外科杂志, 2007, 16(8): 778-782.
[11] CHEN G, WANG Y, ZHOU M, et al. EphA1 receptor silencing by small interfering RNA has antiangiogenic and antitumor efficacy in hepatocellular carcinoma[J]. Oncol Rep, 2010, 23(2): 563-570.
[12] 陈钢, 王怡, 周蒙滔, 等. 抑制PI3K/Akt信号传导对Ephrin-A1介导的肝癌细胞运动和侵袭能力的影响[J]. 中华普通外科杂志, 2009, 24(10): 788-791.
[13] ZHU Z, CHEN G, LI X, et al. Endothelial progenitor cells homing to orthotopic implanted liver tumor of nude mice[J]. J Huazhong Univ Sci Technolog Med Sci, 2012, 32(5): 675-679.
[14] 朱智, 易继林, 陈钢, 等. EphrinA1在肝细胞癌患者内皮祖细胞中的表达及其临床意义[J]. 临床外科杂志, 2011, 19(11): 742-744.