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目的探讨肝癌衍生生长因子(HDGF)和血管内皮生长因子(VEGF)在脑胶质瘤中的表达及其与肿瘤微血管密度(MVD)的相关性。方法选取2008年3月—2013年6月于右江民族医学院附属医院诊治的脑胶质瘤患者52例为脑胶质瘤组,另选取同期于本院行颅脑损伤内减压术患者52例为对照组。收集脑胶质瘤组肿瘤组织和对照组正常脑组织,冷冻保存。实时荧光定量反转录PCR法测定HDGF、VEGF mRNA表达水平。将肿瘤组织和正常脑组织裂解后提取蛋白,分别采用酶联免疫吸附试验(ELISA)和Western blotting测定HDGF、VEGF表达水平。以CD105作为血管内皮细胞的特异性标志物,显微镜下观察5个视野内微血管数计算MVD。自病理检查确诊起,追踪随访观察脑胶质瘤患者至2015-07-30,随访终点为死亡。结果脑胶质瘤组HDGF、VEGF mRNA表达水平高于对照组,经ELISA、Western blotting测定的HDGF、VEGF表达水平均高于对照组,差异有统计学意义(P<0.05)。脑胶质瘤组MVD为(74.3±10.4)个/视野,高于对照组的(9.6±1.4)个/视野,差异有统计学意义(t=15.941,P<0.001)。脑胶质瘤组HDGF mRNA、蛋白(分别经ELISA、Western blotting测定)表达水平与MVD呈正相关(r=0.562、0.382、0.225,P<0.05),VEGF mRNA、蛋白(分别经ELISA、Western blotting测定)表达水平与MVD呈正相关(r=0.676、0.471、0.184,P<0.05)。HDGF表达低水平、高水平的脑胶质瘤患者中位生存时间分别为13.6〔95%CI(11.0,16.1)〕、8.4〔95%CI(6.6,10.2)〕个月,其生存曲线比较,差异有统计学意义(χ~2=9.938,P=0.002)。VEGF表达低水平、高水平的脑胶质瘤患者中位生存时间分别为14.3〔95%CI(12.2,16.8)〕、12.6〔95%CI(10.0,15.1)〕个月,其生存曲线比较,差异无统计学意义(χ~2=0.735,P=0.391)。MVD低水平、高水平的脑胶质瘤患者中位生存时间分别为14.8〔95%CI(12.3,17.3)〕、10.4〔95%CI(8.1,12.6)〕个月,其生存曲线比较,差异有统计学意义(χ~2=4.307,P=0.038)。结论脑胶质瘤组织中HDGF、VEGF表达水平异常升高与肿瘤微血管形成紧密相关,可作为判断肿瘤血管生成和疾病预后的参考标志物。
Objective To investigate the expression of hepatocellular carcinoma-derived growth factor (HDGF) and vascular endothelial growth factor (VEGF) in gliomas and its relationship with tumor microvessel density (MVD). Methods From March 2008 to June 2013, 52 glioma patients diagnosed and treated in the Youjiang Medical College Affiliated Hospital were selected as glioma group. In the same period, 52 cases as control group. The brain tissue of glioma group and normal brain tissue were collected and cryopreserved. Real-time fluorescence quantitative reverse transcription PCR was used to detect the expression of HDGF and VEGF mRNA. The tumor tissue and normal brain tissue were lysed and the protein was extracted. The expression of HDGF and VEGF were detected by enzyme-linked immunosorbent assay (ELISA) and Western blotting respectively. Using CD105 as a specific marker of vascular endothelial cells, MVD was calculated under 5 microscopic visual fields. Since the diagnosis of pathological examination, follow-up observation of glioma patients until 2015-07-30, the end of follow-up was death. Results The expression of HDGF and VEGF mRNA in glioma group was higher than that in control group. The levels of HDGF and VEGF in glioma group were higher than those in control group (P <0.05). The MVD in glioma group was (74.3 ± 10.4) / field, which was higher than that in control group (9.6 ± 1.4) / field, the difference was statistically significant (t = 15.941, P <0.001). The expression of HDGF mRNA and protein (respectively by ELISA and Western blotting) in glioma group was positively correlated with MVD (r = 0.562,0.382,0.225, P <0.05), VEGF mRNA and protein (respectively by ELISA and Western blotting ) Was positively correlated with MVD (r = 0.676,0.471,0.184, P <0.05). The median survival time of HDGF patients with low-grade and high-grade gliomas were 13.6 (95.0% CI, 11.0, 16.1) and 8.4% (95% CI 6.6, 10.2) The difference was statistically significant (χ ~ 2 = 9.938, P = 0.002). The median survival time was 14.3 [95% CI (12.2, 16.8)] and 12.6 [95% CI (10.0, 15.1)] respectively in patients with low-grade and high-grade gliomas. The difference was not statistically significant (χ ~ 2 = 0.735, P = 0.391). The median survival time was 14.8 [95% CI (12.3, 17.3)] and 10.4 [95% CI (8.1, 12.6)] months in MVD patients with low and high grade glioma, respectively. There was statistical significance (χ ~ 2 = 4.307, P = 0.038). Conclusions The abnormal expression of HDGF and VEGF in glioma tissue is closely related to the formation of tumor microvessel, which can be used as a reference marker to judge the tumor angiogenesis and prognosis.