Evaluation of stromal metalloproteinases and vascular endothelial growth factors in a spontaneous metastasis model

This study aims to investigate MMP2 and MT1-MMP protein as well as VEGF-C and VEGF-D mRNA expression in tumor cells and distant organs considered to be targets for metastasis in a tumor spontaneous metastasis model previously described. Cultured tumor cells, able to express pro-MMP2, MMP2, pro-MMP9, and MT1-MMP, develop tumor growth and metastasis, mainly in the liver and spleen, when they are injected in the mammary pad gland of Wistar rats. Immunohistochemical studies of tumor masses showed small groups of tumor cells staining for MT1-MMP but not for MMP2. In the liver, tumor metastatic foci and a stromal positive staining for both MMP2 and MT1-MMP were shown. The spleen and lymph nodes, with only scattered metastatic cells, did not show MMPs immunostaining. Using RT-PCR, a significantly higher VEGF-C and VEGF-D gene expression was shown in the liver of tumor-bearing rats respect to normal rats, whereas spleen and lymph nodes did not show significant differences in mRNA VEGF-C/D levels. Taken together, our results suggest that the stroma microenvironment of target organs for metastasis has the ability to produce MMPs and VEGFs that facilitate the anchorage of tumor cells and promote tumor cell growth and angiogenesis.     

胃癌组织中类肝素酶mRNA表达及与VEGF-D的关系

目的：探讨胃癌组织中类肝素酶、VEGF—D表达及其与胃癌临床病理参数及血管、淋巴管生成的关系。方法：采用逆转录聚合酶链反应（RT—PCR）法及免疫组织化学方法分别检测48例胃癌及癌旁组织中类肝素酶mRNA和VEGF—D的表达。结果：类肝素酶在胃癌组织中的阳性表达显著高于癌旁组织（P〈0．05）。在高中分化、Ⅰ～Ⅱ期的胃癌组织中类肝素酶的表达明显比低分化、Ⅲ～Ⅳ期减低（P〈0．01）。有淋巴结转移组类肝素酶表达明显比无淋巴结转移组增高（P〈0．01）。VEGF-D在胃癌组织中表达亦显著高于癌旁组织（P〈0．05），且亦与淋巴结转移有关（P〈0．05）。在类肝素酶阳性表达组VEGF—D表达显著高于类肝素酶阴性表达组（P〈0．05）。结论：类肝素酶促进了胃癌的生长、浸润及转移和血管、淋巴管生成，可作为胃癌预后的参考指标。     

VEGF-C、VEGF-D及其受体FLt-4与大肠癌淋巴转移及预后的相关性研究

目的:研究VEGF-C、VEGF-D及其受体Flt-4在大肠癌组织中的表达规律,探讨其与大肠癌浸润特点、淋巴转移等生物学特性及预后的关系。方法:采用免疫组织化学染色法检测45例大肠癌组织中VEGF-C、VEGF-D及Flt-4表达情况,评估VEGF-C、VEGF-D及Flt-4的表达与大肠癌生物学特性及预后的关系。结果:研究发现大肠癌组织VEGF-C和VEGF-D及其受体Flt-4阳性表达率显著高于正常组织(P<0.05)。并且VEGF-C/VEGF-D表达与Flt-4蛋白表达呈显著正相关(P<0.01)。VEGF-C、VEGF-D及Flt-4的表达在有无淋巴管浸润组、有无淋巴结转移组及不同Dukes分期中亦有显著性差异(P<0.05),45例大肠癌患者术后5年生存率为53.3%。其中VEGF-C、VEGF-D和Flt-4阳性患者5年生存率均分别低于阴性患者组(P<0.05)。结论:在大肠癌组织中VEGF-C和VEGF-D与其受体Flt-4的表达与大肠癌淋巴管的发生及促进肿瘤细胞经淋巴管转移可能起一定作用,VEGF-C和VEGF-D表达的检测可作为预测大肠癌淋巴结转移的辅助指标;VEGF-C和VEGF-D蛋白高表达与大肠癌患者预后相关,可能成为今后大肠癌预后评估的分子指标之一。     

食管鳞癌中VEGF-C、VEGF-D的表达与临床病理特征的相关性研究

目的 检测血管内皮生长因子-C、D(VEGF-C和VEGF-D)在食管鳞癌中的表达与各临床病理特征之间的关系,探索食管鳞癌淋巴结转移的机制.方法 免疫组化法检测VEGF-C、D在62例食管鳞癌根治术后标本及20例癌旁正常组织中的表达,计数微淋巴管密度(MLD).分析VEGF-C、D表达及MLD与临床病理特征之间的关系.结果 肿瘤组织中VEGF-C和VEGF-D的高表达率分别是74%(46/62)和82%(51/62);肿瘤边缘MLD(5.387±4.986)明显高于癌旁正常组织(1.900±2.245)(P<0.05).VEGF-C、D高表达组与低表达组相比,淋巴结转移更常见、临床分期也较晚,MLD增加与VEGF-C、D高表达、有淋巴结转移及临床分期较晚有关.VEGF-C和VEGF-D的表达有正相关关系(r=0.691,P<0.001).结论 VEGF-C和VEGF-D在食管鳞癌中可能通过增加肿瘤周边区域的淋巴管新生促进肿瘤的淋巴道转移.     

VEGF-D在口腔鳞癌相关成纤维细胞的表达研究

目的:研究VEGF-D在口腔癌相关成纤维细胞(carcinoma-associated fibroblasts,CAFs)的表达情况以及其表达水平是否与口腔鳞癌淋巴结转移相关。方法:通过免疫组化和RT-PCR检测VEGF-D在NFs、无淋巴结转移CAFs、淋巴结转移CAFs的蛋白和mRNA表达情况。结果:成功培养和纯化CAFs,与NFs相比,CAFs阳性表达α-SMA,生长增殖速度明显增加,细胞排列混乱;VEGF-D在淋巴结转移CAFs的染色强度明显增加,用2-ΔΔCT法计算VEGF-D的mRNA表达水平分别为0.806 7±0.117、1.192 5±0.125、2.085 3±0.131。结论:与NFs相比,CAFs表达VEGF-D明显增加,且与患者淋巴结转移相关。     

乳腺癌中ErbB2过表达与血管内皮生长因子A,C和D高表达的关系

目的:探讨在乳腺癌中ErbB2过表达与血管内皮生长因子A,C和D的高表达关系。方法:用免疫组织化学方法检测107例乳腺癌患者和22例良性肿瘤中ErbB2、VEGF-A、VEGF-C和VEGF-D的表达。结果:VEGF-C和VEGF-D在乳腺癌的表达率高于乳腺的良性肿瘤,然而,VEGF-A、VEGF-C和VEGF-D的表达与erbB2的表达呈显著正相关,VEGF-A高表达而生存率降低,乳腺癌患者中VEGF-C或者VEGF—D高表达时,生存率更低,但是没有统计学意义,然而VEGF-A和VEGF—C联合表达水平比单一表达更有统计学意义。结论:血管与淋巴管的表达与ErbB2有一定的关系,VEGF-A和VEGFC的同时表达对患者预后生存率更有价值。     

Lymphangiogenesis induced by VEGF-C and VEGF-D promotes metastasis and a poor outcome in breast carcinoma: a retrospective study of 61 cases

Purpose To evaluate lymphangiogenesis in patients with breast carcinoma, explore the underlying mechanism, and study the relationship between lymphangiogenesis and progression of breast carcinoma. Methods Sixty-one cases of breast carcinoma with complete clinical and pathological data were analyzed. Using an anti-podoplanin monoclonal antibody, an immunohistochemical study was made of all specimens to detect lymphatic vessel density (LVD) and to investigate its clinicopathological and prognostic value. VEGF-C and VEGF-D were observed by RT-PCR and immunostaining to investigate their clinicopathological and prognostic values and their relationship with lymphangiogenesis. Results LVD in breast carcinoma (6.28 ± 3.73) was significantly higher than in benign mammary lesions (0.50 ± 1.27), P < 0.01 and was significantly associated with lymphatic metastasis and high TNM stage, P < 0.01. The level of VEGF-C and VEGF-D expression was also significantly higher in breast carcinomas than in benign mammary lesions, P < 0.01. LVD increased significantly with higher expression of VEGF-C and VEGF-D, P < 0.01. Patients with high expression of VEGF-C and VEGF-D were observed to be more likely to have a bad outcome, P < 0.05. Conclusions Lymphangiogenesis was significantly associated with lymph node metastasis, high TNM, and poor outcome in breast carcinoma. LVD may serve as a predictor of lymph node metastasis and a prognostic factor in breast carcinoma. VEGF-C and VEGF-D play an important role in lymphangiogenesis making the carcinoma more aggressive and leading to a poor prognosis.     

结肠癌组织ＨＭＧＢ１和ＶＥＧＦ-Ｄ的表达及其相关性

目的：探讨高迁移率族蛋白１（ＨＭＧＢ１）和血管内皮生长因子Ｄ（ＶＥＧＦ-Ｄ）在人结肠癌组织中的表达及其二者之间的相关性。方法：收集２００３～２００５年手术切除的７０例结肠癌组织标本,用免疫组织化学染色法检测结肠癌组织ＨＭＧＢ１和ＶＥＧＦ-Ｄ的表达情况,同时对两者行相关性分析。ＨＭＧＢ１作为干预因素,检测经ＨＭＧＢ１刺激后结肠癌细胞ＨＣＴ１１６中ＶＥＧＦ-ＤｍＲＮＡ表达的变化。结果：ＨＭＧＢ１在结肠癌组织中的阳性表达率为７２.９％,其中伴有淋巴结转移的表达率为８３３％,无淋巴结转移的表达率为５７.１％,两者差异有统计学意义。ＶＥＧＦ-Ｄ在结肠癌组织中的阳性表达率为３０％。ＨＭＧＢ１表达与ＶＥＧＦ-Ｄ表达呈正相关。结肠癌细胞ＨＣＴ１１６中ＶＥＧＦ-ＤｍＲＮＡ的表达受ＨＭＧＢ１的影响。结论：ＨＭＧＢ１可能通过诱导结肠癌组织中ＶＥＧＦ-Ｄ的表达,从而促进结肠癌的淋巴结转移。