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目的探讨脯氨酸羟化酶2(PHD2)蛋白在肝细胞癌(HCC)血管生成中的作用。方法 (1)采用免疫组织化学S-P法,检测72例肝细胞癌组织中的PHD2表达,结合肝细胞癌临床病理和MVD资料加以分析。(2)以不同浓度二氯化钴(CoCl2)诱导体外培养微血管内皮细胞(HMEC-1)缺氧。Western-blot方法检测PHD2、HIF-1α、VEGF蛋白表达水平,基质胶内皮细胞二维成管实验检测HMEC-1细胞的成管能力。结果 PHD2在72例HCC组织中的阳性表达率22.2%(16/72),低于在癌旁肝组织中阳性表达率68.1%(49/72)。HCC组织中PHD2高表达组MVD计数(52.1±4.3)显著性低于PHD2低表达组(69.2±5.5)。CoCl2处理后HMEC-1细胞PHD2蛋白表达下调,HIF-1α蛋白降解减少,VEGF表达上调,HMEC-1细胞成管能力下降,与对照组(正常肝组织)比较差异具有统计学意义(P<0.05)。结论缺氧微环境下,PHD2表达水平下调,内皮细胞增殖,但细胞成管能力下降,是导致HCC瘤体血管成熟度下降,促进HCC侵袭转移的重要影响因素之一。
Objective To investigate the role of prolyl hydroxylase 2 (PHD2) in angiogenesis of hepatocellular carcinoma (HCC). Methods (1) The expression of PHD2 in 72 cases of hepatocellular carcinoma was detected by immunohistochemical S-P method, and combined with clinicopathological and MVD data of hepatocellular carcinoma. (2) Induced hypoxia of cultured microvascular endothelial cells (HMEC-1) with different concentrations of cobalt chloride (CoCl2). The protein expression of PHD2, HIF-1α and VEGF was detected by Western-blot and the tube formation ability of HMEC-1 cells was detected by two-dimensional tube formation assay. Results The positive expression rate of PHD2 in 72 HCC tissues was 22.2% (16/72), which was lower than that in para-cancerous liver tissues (68.1%, 49/72). The MVD count (52.1 ± 4.3) in HCC tissues with PHD2 high expression group was significantly lower than that in PHD2 low expression group (69.2 ± 5.5). Compared with the control group (normal liver tissue), the difference of PHD2 protein expression, the decrease of HIF-1α protein expression, the increase of VEGF expression and the ability of tube formation of HMEC-1 cells in CoCl2 treatment group were statistically significant (P <0.05 ). Conclusion Under hypoxic conditions, the expression of PHD2 is down-regulated and endothelial cells proliferate, but the cell-forming ability is decreased. This is one of the important factors that lead to the decrease of vascular maturity and the invasion and metastasis of HCC.