SP1介导CD147通过HIF-1α/VEGF信号通路影响卵巢癌的血管形成

SP1 mediates CD147 to influence the angiogenesis of ovarian cancer through HIF-1α/VEGF signaling pathway

摘要:

目的： 探讨特异性蛋白（specific protein，SP1）介导CD147，即细胞外基质金属蛋白诱导因子（extracellular matrix metalloprotein inducer，EMMPRIN）的表达激活缺氧诱导因子-1α（hypoxia inducible factor-1α，HIF-1α）/血管内皮生长因子（vascular endothelial growth factor，VEGF）信号通路对卵巢癌血管形成的影响。方法： 血管生长是肿瘤发生的关键步骤，多数恶性肿瘤的血管生成密集且生长迅速。这是由于肿瘤细胞自身可分泌多种生长因子，诱导血管生成。因此，血管生成在肿瘤的发展转移过程中起到重要作用，抑制这一过程将能明显阻止肿瘤组织的发展和扩散转移。体外的血管生成试验能很好地模拟肿瘤的血管发生过程，实验分为过表达组（OE组）、沉默组（SI组）、控制组（CONTROL组）、阴性对照质粒组（NC组），通过沉默或过表达卵巢癌细胞株SKOV3中SP1或CD147，并使用HIF-1α蛋白翻译抑制剂KC7F2阻断HIF-1α/VEGF信号通路，然后用培养上清液作用于人脐静脉血管内皮细胞HUVEC-12，进行体外血管形成实验来模拟肿瘤血管生成改变，并使用荧光实时定量PCR技术检测CD147和VEGF在细胞中的表达分布。结果： 阴性对照组血管分支数（206.33±16.03）高于SI-CD147组血管分支数（137.00±15.09）；OE-SP1组的CD147 mRNA水平（2.38±0.26）高于SI-SP1组中CD147 mRNA水平（0.55±0.12）；SI-SP1+OE-CD147组血管分支数（200.00±19.52）高于SI-SP1组血管分支数（130.00±12.01）；OE-CD147组血管分支数（373.67±29.02）高于OE-CD147+KC7F2组血管分支数（187.00±19.52），OE-CD147组VEGF分子水平（4.05±0.09）高于OE-CD147+KC7F2组VEGF分子水平（1.02±0.11）。结论： 卵巢癌中CD147表达升高，上游转录因子SP1可以上调CD147表达，从而激活HIF-1α/VEGF信号通路，促进卵巢癌血管形成过程，最终参与卵巢癌发病。

Abstract:

Objective: To discuss the expression of CD147 [extracellular matrix metalloprotein inducer (EMMPRIN)]mediated by specific protein (SP1), the activation of hypoxia inducible factor-1α (HIF-1α), and the effect of vascular endothelial growth factor (VEGF) signaling pathway on angiogenesis in ovarian cancer. Methods: The angiogenesis is the critical step in tumorigenesis. Most malignant tumors have dense angiogenesis and grow fast, which is due to the tumor cells can secrete many kinds of growth factors themselves to induce angiogenesis. Therefore, angiogenesis plays an important role in the development and metastasis of tumors and inhibition of it will significantly prevent the development and metastasis of tumor tissues.In vitro angiogenesis assay could simulate the procession of tumor angiogenesis well. In this study, the experiment was divided into overexpression group (OE group), silencing group (SI group), CONTROL group (CONTROL group), and negative CONTROL group (NC group). By silencing or over-expressing of SP1 or CD147 in ovarian cancer cell line SKOV3, blocking HIF-1α/VEGF signal pathway with HIF-1α protein translation inhibitor KC7F2, and then using culture supernatants to act on human umbilical vein endothelial cells (HUVEC-12), in vitro angiogenesis assay was carried out to simulate tumor angiogenesis changes. Meanwhile, fluorescence real-time quantitative PCR (RT-qPCR) was used to examine the expressions of CD147 and VEGF in the cells. Results: The amount of vascular arborizations in NC group (206.33±16.03) was more than that in SI-CD147 group (137.00±15.09). The mRNA level of CD147 in OE-SP1 group (2.38±0.26) was higher than that in SI-SP1 group (0.55±0.12). The amount of vascular arborizations in SI-SP1+OE-CD147 group (200.00±19.52) was more than that in SI-SP1 group (130.00±12.01). The amount of vascular arborizations in OE-CD147 group (373.67±29.02) was more than that in OE-CD147+KC7F2 group (187.00±19.52). The molecular level of VEGF in OE-CD147 group (4.05±0.09) was higher than that in OE-CD147+KC7F2 group (1.02±0.11). Conclusion: The expression of CD147 increases in ovarian cancer, and the upstream transcription factor SP1 is able to upregulate the expression of CD147, thereby activating HIF-1α/VEGF signaling pathway to promote the angiogenesis of ovarian cancer, and ultimately leading to the pathogenesis of ovarian cancer.