Role of hypoxia-inducible factor 1α in gastric cancer cell growth, angiogenesis, and vessel maturation
Role of hypoxia-inducible factor 1α in gastric cancer cell growth, angiogenesis, and vessel maturation
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DOI：
10.1093/jnci/djh168
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发表时间：
2004-06-16
影响因子：
10.3
通讯作者：
Ellis, LM
中科院分区：
文献类型：
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作者：
Stoeltzing, O;McCarty, MF;Ellis, LM
背景资料：缺氧诱导因子1（HIF-1）是一种由氧调节亚基、HIF-1 α和HIF-1 β组成的异源二聚体，介导血管内皮生长因子（VEGF）基因的转录。HIF-α的过表达与肿瘤血管生成和肿瘤细胞增殖和侵袭有关。我们研究了抑制HIF-1 α活性对血管生成和人胃癌生长的影响。研究方法：人胃癌TMK-1细胞用pHIF-1alphaDN或空表达载体（pCEP 4）稳定转染，pHIF-1alphaDN是编码显性负性形式的HIF-1 α的表达质粒，其与内源性HIF-1 β二聚化以产生不能激活转录的HIF-1复合物。在所有实验中测试pHIF-1 α DN转染的细胞的两个克隆，DN 2和DN 3。我们使用酶联免疫吸附试验来测量VEGF分泌转染细胞培养在缺氧（1%O-2）或非缺氧（20%O-2）条件下。我们使用皮下和原位小鼠肿瘤模型来检查来自注射pHIF-1 α DN或pCEP 4转染细胞的肿瘤的生长。通过免疫组化或免疫荧光染色分析肿瘤细胞增殖、血管面积（功能性血管体积的测量）和肿瘤内皮细胞与周细胞样细胞的相关性（血管成熟的测量）。所有统计检验均为双侧检验。结果：当在非缺氧或缺氧条件下培养时，DN 2细胞和DN 3细胞比pCEP 4转染的TMK-1细胞分泌较少的VEGF（例如，在非缺氧条件下，DN 2与pCEP 4：645 pg VEGF/10（6）细胞与1591 pg VEGF/10（6）细胞，差异= 946 pg VEGF/10（6）细胞[95%置信区间{CI} = 640至1251 pg VEGF/10（6）细胞; P =.006];在缺氧条件下，DN 2与pCEP 4：785 pg VEGF/10（6）细胞vs 2807 pg VEGF/10（6）细胞，差异= 2022 pg VEGF/10（6）细胞[95% CI = 1871 - 2152 pg VEGF/10（6）细胞; P
Background: Hypoxia-inducible factor 1 (HIF-1), a heterodimer comprising the oxygen-regulated subunit, HIF-1alpha, and HIF-1beta, mediates transcription of the gene for vascular endothelial growth factor (VEGF). Overexpression of HIF-alpha is associated with tumor angiogenesis and tumor cell proliferation and invasion. We examined the effects of inhibiting HIF-1alpha activity on angiogenesis and human gastric cancer growth in vivo. Methods: Human gastric cancer TMK-1 cells were stably transfected with pHIF-1alphaDN, an expression plasmid encoding a dominant-negative form of HIF-1alpha that dimerizes with endogenous HIF-1beta to produce HIF-1 complexes that cannot activate transcription, or with the empty expression vector (pCEP4). Two clones of pHIF-1alphaDN-transfected cells, DN2 and DN3, were tested in all experiments. We used an enzyme-linked immunosorbent assay to measure VEGF secretion by transfected cells cultured in hypoxic (1% O-2) or nonhypoxic (20% O-2) conditions. We used subcutaneous and orthotopic mouse tumor models to examine the growth of tumors derived from injected pHIF-1alphaDN- or pCEP4-transfected cells. Tumor cell proliferation, vessel area (a measure of functional vascular volume), and tumor endothelial cell association with pericyte-like cells (a measure of vessel maturation) were analyzed by immunohistochemical or immunofluorescent staining. All statistical tests were two-sided. Results: DN2 cells and DN3 cells secreted less VEGF than pCEP4-transfected TMK-1 cells when cultured in nonhypoxic or hypoxic conditions (e.g., DN2 versus pCEP4 in nonhypoxic conditions: 645 pg of VEGF/10(6) cells versus 1591 pg of VEGF/10(6) cells, difference = 946 pg of VEGF/10(6) cells [95% confidence interval {CI} = 640 to 1251 pg of VEGF/10(6) cells; P =.006]; DN2 versus pCEP4 in hypoxic conditions: 785 pg of VEGF/10(6) cells versus 2807 pg of VEGF/10(6) cells, difference = 2022 pg of VEGF/10(6) cells [95% CI = 1871 to 2152 pg of VEGF/10(6) cells; P