摘要目的:检测子宫内膜癌中VEGF-C、VEGF-D的表达及D2-40标记的淋巴管密度的关系,探讨其临床意义。方法:收集52例子宫内膜癌术后存档的石蜡标本作为实验组,选取40例增生期子宫内膜标本作为对照组,应用免疫组织化学技术检测二组中VEGF-C、VEGF-D及D2-40标记的淋巴管密度(MLVD)的表达。结果:子宫内膜癌中VEGF-C、VEGF-D表达的阳性率明显高于对照组(P<0.05),实验组中D2-40标记的微淋巴管密度明显高于对照组(P<0.05),VEGF-C、VEGF-D及MLVD与子宫内膜癌的浸润深度及淋巴结转移密切相关(P<0.05)。结论:子宫内膜癌中存在D2-40阳性的淋巴管新生,VEGF-C及VEGF-D不仅参与淋巴管新生,还可能与患者预后相关。

    关键词子宫内膜癌;VEGF-C;VEGF-D;淋巴管;D2-40;免疫组织化学

    AbstractObjective:To detect the expressions of VEGF-C, VEGF-D and D2-40 in endometrial carcinoma, and investigate their relationship with lymphangiogenesis in different clinical characteristic of neoplasm. Methods: The study included 52 cases of endometrial carcinoma as experimental group and 40 cases of proliferative endometrium tissues as control group. We analyzed their expressions of VEGF-C, VEGF-D and D2-40 in different pathological character by using the IHC. Results: Comparisons of the primary tumors with endometrial carcinoma and proliferative endometrium tissues showed that there were significant high-regulation of VEGF-C, VEGF-D and LMVD(P<0.05). The expressions of VEGF-C, VEGF-Dand LMVD were associated with infiltration degree and lymph node metastasis(P<0.05). There was positive correlationship between VEGF-C and VEGF-D, VEGF-C and LMVD, VEGF-D and LMVD in endometrial carcinoma(P<0.05). Conclusions:VEGF-C and VEGF-D may promote the lymphangiogenesis of positive D2-40. The combined examination of VEGF-C and VEGF-D may be helpful to predict the prognosis in patients with endometrial carcinoma. ......
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     摘要目的:检测子宫内膜癌中VEGF-C、VEGF-D的表达及D2-40标记的淋巴管密度的关系,探讨其临床意义。方法:收集52例子宫内膜癌术后存档的石蜡标本作为实验组,选取40例增生期子宫内膜标本作为对照组,应用免疫组织化学技术检测二组中VEGF-C、VEGF-D及D2-40标记的淋巴管密度(MLVD)的表达。结果:子宫内膜癌中VEGF-C、VEGF-D表达的阳性率明显高于对照组(P<0.05),实验组中D2-40标记的微淋巴管密度明显高于对照组(P<0.05),VEGF-C、VEGF-D及MLVD与子宫内膜癌的浸润深度及淋巴结转移密切相关(P<0.05)。结论:子宫内膜癌中存在D2-40阳性的淋巴管新生,VEGF-C及VEGF-D不仅参与淋巴管新生,还可能与患者预后相关。

    关键词子宫内膜癌;VEGF-C;VEGF-D;淋巴管;D2-40;免疫组织化学

    AbstractObjective:To detect the expressions of VEGF-C, VEGF-D and D2-40 in endometrial carcinoma, and investigate their relationship with lymphangiogenesis in different clinical characteristic of neoplasm. Methods: The study included 52 cases of endometrial carcinoma as experimental group and 40 cases of proliferative endometrium tissues as control group. We analyzed their expressions of VEGF-C, VEGF-D and D2-40 in different pathological character by using the IHC. Results: Comparisons of the primary tumors with endometrial carcinoma and proliferative endometrium tissues showed that there were significant high-regulation of VEGF-C, VEGF-D and LMVD(P<0.05). The expressions of VEGF-C, VEGF-Dand LMVD were associated with infiltration degree and lymph node metastasis(P<0.05). There was positive correlationship between VEGF-C and VEGF-D, VEGF-C and LMVD, VEGF-D and LMVD in endometrial carcinoma(P<0.05). Conclusions:VEGF-C and VEGF-D may promote the lymphangiogenesis of positive D2-40. The combined examination of VEGF-C and VEGF-D may be helpful to predict the prognosis in patients with endometrial carcinoma.

    Key Wordsendometrial carcinoma;VEGF-C;VEGF-D;lymphangiogenesis;D2-40;IHC

    中图分类号:R737.33文献识别码:A

    子宫内膜癌发展过程中淋巴道转移是其重要组成部分。血管内皮生长因子-C(vascular endothelial growth factor-C, VEGF-C)和血管内皮生长因子-D(vascular endothelial growth factor-C, VEGF-D)是新发现的,被认为是与淋巴管生成有关的抗体[1],而D2-40也是新发现的最特异的淋巴管标记物。本实验应用免疫组织化学技术检测子宫内膜癌中VEGF-C、VEGF-D及D2-40的表达,探讨VEGF-C、VEGF-D与D2-40标记的淋巴管密度(micro1ymphatic vessel density, MLVD)的关系,以期为研究肿瘤的生物学行为提供帮助。

    1资料与方法

    1.1 临床资料

    收集我院2005年1月至2009年6月子宫内膜癌手术后病理科存档的蜡块,所有标本均经病理主治医师再次确诊。本组共52例,其中绝经前12例,绝经后40例,年龄41～68岁,平均58.1岁。排除标准:①术前经抗癌治疗的患者;②合并其它器官恶性肿瘤的患者。同时选取诊刮后病理证实为增生期子宫内膜的标本40例作为对照。

    1.2 免疫组织化学方法

    实验用VEGF-C、VEGF-D及D2-40抗体均购自北京中杉金桥生物技术有限公司。免疫组织化学染色采用S-P法,具体步骤:具体步聚:脱蜡,水化组织切片,3%H2O2去离子水孵育10分钟,以阻断内源性过氧化物酶;抗原修复后分别滴加一抗,室温37度孵育1～2小时或过夜,PBS冲洗,5分钟×3次;滴加生物素标记的二抗,室温37度孵育20分钟,PBS冲洗,5分钟×3次;滴加辣根酶标记链霉卵白素,室温37度孵育20～30分钟,PBS冲洗,5分钟×3次;应用DAB显色剂显色;自来水充分冲洗、复染、脱水、透明、封片。本实验中所有标本的检测均由同一病理技师完成,并在显微镜下显色观察,严格质控。实验中设用PBS代替一抗作为阴性对照。

    1.3 结果判定标准

    免疫组化染色结果:VEGF-C、VEGF-D蛋白阳性染色为棕黄色颗粒样着色,均定位于细胞浆。随机选取10个高倍视野(400倍),计算每个视野内阳性染色细胞的百分数,并取平均值。肿瘤内微淋巴管密度计数:低倍镜下(40倍)首先排除肿瘤出血区及边缘区,每张切片中选择5个淋巴管密度最高的区域,然后在高倍镜下(400倍)统计每个区域的微淋巴管数目,取平均值即为微淋巴管密度。

    1.4 统计学方法

    实验数据应用SAS统计软件进行统计分析,定性资料两组间比较采用卡方检测,定量资料两组间比较采用t检验,并进行线性相关分析,以P<0.05为差异有统计学意义。

    2 结果

    2.1 VEGF-C、VEGF-D及LMVD在实验组与对照组中的表达

    实验组中VEGF-C和VEGF-D表达的阳性率明显高于对照组,实验组中微淋巴管计数明显高于对照组(见表1)。

    2.2 VEGF-C、VEGF-D及LMVD的表达与子宫内膜癌临床病理特征的关系

    实验组中VEGF-C、VEGF-D表达的阳性率、微淋巴管密度均与肿瘤的浸润深度及淋巴结转移有关(见表2)。

    2.3 VEGF-C、VEGF-D及LMVD在子宫内膜癌中表达的关系

    对子宫内膜癌组三种蛋白的表达进行线性相关分析,结果显示子宫内膜癌中VEGF-C与VEGF-D的表达呈正相关(r=0.36, P<0.05)。VEGF-C与LMVD呈正相关(r=0.42, P<0.05)。VEGF-D与LMVD呈正相关(r=0.45, P<0.05)。