乳腺癌组织中LVD、VEGF-C和MMP-9表达及其相关性

目的 检测乳腺浸润性导管癌(invasive ductal carcinoma,IDC)组织中的淋巴管密度(lymphatic vessel density,LVD)、VEGF-C及MMP-9的表达,探讨其临床意义.方法 收集80例乳腺IDC和20例乳腺增生性病变,应用免疫组化PV-6000两步法检测淋巴管内皮透明质酸受体1(LYVE-1)、VEGF-C和MMP-9的表达.结果 乳腺癌组织中LYVE-1标记的LVD、VEGF-C和MMP-9的表达明显高于乳腺增生性病变,差异均有统计学意义(P<0.01).LVD、VEGF-C与临床病理参数之间均无差异(P>0.05).MMP-9的表达水平在肿瘤直径>2 cm组和淋巴结转移阳性组明显升高(P<0.05和P<0.01);而在组织学分级之间表达的差异无统计学意义(P>0.05).LVD与VEGF-C的表达呈正相关关系(r=0.398,P<0.01);LVD和MMP-9之间无相关关系(P>0.05).结论 IDC中淋巴管生成增多,VEGF-C是乳腺癌中淋巴管生成的重要刺激因子,促进乳腺癌中淋巴管的生成;但乳腺癌淋巴管密度与淋巴道转移无关.MMP-9参与肿瘤的侵袭和转移的过程,促进乳腺癌淋巴道转移.     

抑癌基因PTEN和血管内皮生长因子-C表达与乳腺癌淋巴管密度的相关性及其临床意义

目的:研究乳腺癌中抑癌基因PTEN和血管内皮生长因子-C(VEGF-C)表达与肿瘤间质淋巴管密度(LVD)的相关性及其临床意义。方法:应用EliVision免疫组化法检测90例乳腺浸润性导管癌组织和30例乳腺纤维腺瘤组织中PTEN、VEGF-C和淋巴管内皮细胞透明质酸受体1(LYVE-1)的表达,计算LYVE-1标记的乳腺癌间质淋巴管的密度,分析PTEN和VEGF-C与LVD的关系。结果:PTEN和VEGF-C在乳腺癌中的表达阳性率分别为48.9%、47.8%,LVD为(8.03±2.26)个/HPF,与良性对照组比较均有显著差异(P<0.01)。PTEN阳性率随乳腺癌淋巴结转移的出现和临床分期的升高而降低(P<0.05),乳腺癌VEGF-C阳性率随淋巴结转移的出现和临床分期的升高而增加(P<0.05),PTEN阳性组乳腺癌的LVD水平低于PTEN阴性组(P<0.05),VEGF-C阳性组乳腺癌的LVD水平高于VEGF-C阴性组(P<0.01),PTEN阳性组乳腺癌的VEGF-C阳性率低于PTEN阴性组(P<0.05)。结论:PTEN和VEGF-C的异常表达与乳腺癌间质淋巴管密度密切相关,在乳腺癌的淋巴管生成及侵袭转移过程中具有重要作用。     

VEGF-C、VEGFR-3和iNOS在人乳腺癌淋巴管的表达

目的检测乳腺癌组织血管内皮生长因子-C(VEGF-C)及其受体(VEGFR-3)、iNOS基因表达的相关性及这三个基因mRNA表达水平与淋巴管密度(LVD)的相关性,为阐明乳腺癌淋巴管生成的分子机理提供实验依据。方法RT-PCR检测乳腺癌组织及正常乳腺组织VEGF-C、VEGFR-3、iNOSmRNA的表达水平;免疫组织化学染色法检测淋巴管内皮细胞上VEGFR-3的表达,测定淋巴管密度。结果乳腺癌LVD为20.35±4.23,显著高于正常对照组(P<0.05);乳腺癌组织VEGF-C、VEGFR-3、iNOSmRNA三者的表达率分别为74.0%、84.0%、82.0%,显著高于正常对照组(P<0.05);VEGF-C、VEGFR-3、iNOS阳性组中LVD分别为21.34±3.45、18.54±4.68、17.43±4.76,显著高于对照组(P<0.05)。结论VEGF-C、VEGFR-3、iNOSmRNA表达与淋巴管密度之间呈正相关,并且3者之间的表达亦具有相关性,这些基因的表达增高可能在乳腺癌淋巴管生成过程中具有重要作用。     

基质金属蛋白酶2和9及血管内皮生长因子C在乳腺癌淋巴结转移中的作用

目的 研究血管内皮生长因子C(VEGF-C)与基质金属蛋白酶(MMP)-2、MMP-9在乳腺癌组织中的表达及三者在乳腺癌淋巴结转移中的作用.方法 应用免疫组织化学SP法对84例乳腺癌(有腋淋巴结转移者52例,无腋淋巴结转移者32例)的VEGF-C、MMP-2和-9以及血管内皮透明质酸受体-1(LYVE-1)的表达进行检测,统计分析VEGF-C、MMP-2和-9与乳腺癌淋巴管生成的关系.利用重组质粒表达载体介导的短发夹式干扰RNA(shRNA)靶向沉默乳腺癌MCF-7细胞株中VEGF-C基因,PCR检测VEGF-C、MMP-2和-9的表达.结果 VEGF-C与MMP-2和-9在乳腺癌组织中均呈过表达,分别为83.3%(70/84)、89.3%(75/84)和75.0%(63/84),且在腋淋巴结转移组[94.2%(49/52)、98.1%(51/52)和88.5%(46/52)]比无淋巴结转移组[65.6%(21/32)、75.0%(24/32)和53.1%(17/32)]明显高表达(P＜0.05);淋巴管密度在腋淋巴结转移组及无转移组的表达有统计学意义(P＜0.05),随着VEGF-C与MMP-2和-9表达强度增加,淋巴管数量也增加(P＜0.05);转染重组质粒后MCF-7细胞株的VEGF-C及MMP-2和-9的mRNA表达水平下调,抑制率分别为95.0%、53.0%和77.0%(P＜0.05).结论 VEGF-C与MMP-2和-9协同促进乳腺癌组织的淋巴管新生和乳腺癌淋巴结转移.     

VEGF-C及VEGFR-3在乳腺癌中的表达

目的探讨乳腺癌中血管内皮生长因子-C(VEGF-C)、血管内皮生长因子受体-3(VEGFR-3)的表达及与淋巴管生成的关系。方法采用免疫组化SP法检测57例乳腺癌组织中VEGF-C及VEGFR-3的表达,并在显微镜下记数VEGFR-3标记的脉管。结果淋巴结转移组VEGF-C的阳性率(90.48%)显著高于无淋巴结转移组(47.22%)(P<0.05);淋巴结转移组VEGFR-3阳性脉管数(7.62±1.18)显著高于无淋巴结转移组(5.27±0.96)(P<0.05);VEGF-C的阳性表达与VEGFR-3阳性脉管数呈正相关,相关系数为r为0.882(P<0.05)。结论 VEGF-C及VEGFR-3与乳腺癌的生长,淋巴管的生长及淋巴结的转移有关。     

喉癌中血管内皮素-1的表达与血管内皮生长因子-C、淋巴管生成及预后的关系

目的:探讨血管内皮素-1(ET-1)在喉癌组织中的表达与血管内皮生长因子-C(VEGF-C)、淋巴管密度(LVD)、淋巴转移之间的关系及其在喉癌预后中的意义.方法:以45例经病理确诊的喉癌组织为实验组,20例喉良性病变组织为对照组,采用免疫组织化学SP法对上述组织中ET-1、VEGF-C蛋白的表达进行分析,同时采用5′-核苷酸酶-碱性磷酸酶双重染色法(5'-Nase-ALP)计数LVD,并结合临床病理特征和生存资料进行相关分析.结果:喉癌组织中ET-1和VEGF-C的表达均高于良性病变组织,且ET-1和VEGF-C的表达具有相关性;在喉癌组织中ET-1的表达与瘤内LVD和瘤周LVD、淋巴结转移、淋巴管浸润、TNM临床分期呈正相关,ET-1+/VEGF-C+、VEGF-C的表达与生存率呈负相关,其中ET-1+/VEGF-C+组具有高危死亡率.结论:ET-1促进喉癌淋巴管生成和淋巴管转移,ET-1信号途径和VEGF-C信号途径可能对喉癌淋巴管生成和转移有协同促进作用,联合检测ET-1和VEGF-C的表达可成为独立判断喉癌预后的新的生物学指标.     

Smad4和VEGF-C的表达与乳腺癌淋巴管生成之间的关系

目的观察Smad4和血管内皮生长因子(VEGF)-C在人乳腺癌组织内的表达情况,分析Smad4和VEGF-C的表达与乳腺癌淋巴管生成及淋巴结转移之间的关系。方法取乳腺癌病例56例,其中,淋巴结转移组35例,无淋巴结转移组21例。应用免疫组化法和Western blot技术观察Smad4和VEGF-C在乳腺癌组织内的表达。以D2-40作为淋巴管内皮特异性标记物,检测乳腺癌组织内淋巴管生成情况。结果 Smad4在无淋巴结转移组的表达率明显高于其在有淋巴结转移组的表达率,Smad4表达阳性组的淋巴管数密度(LVD)明显低于Smad4表达阴性组。而VEGF-C在淋巴结转移组的表达率明显高于其在无淋巴结转移组的表达率。Smad4的表达与VEGF-C的表达呈显著的负相关性(r=-0.314)。Western blot检测结果表明,VEGF-C蛋白在有淋巴结转移乳腺癌组织中的表达量高于其在无淋巴结转移乳腺癌组织内的表达量,而Smad4在有淋巴结转移乳腺癌组织内的表达量明显低于其在无淋巴结转移组的表达量。结论 VEGF-C在乳腺癌淋巴管的发生及淋巴结转移中发挥重要作用,与Smad4的表达呈负相关,Smad4可能通过调节VEGF-C蛋白的表达而抑制乳腺癌淋巴管生成和淋巴道转移的作用。     

VEGF-C在淋巴管生成及乳腺癌淋巴道转移中的作用

目的： 探讨阻断VEGF-C/Flt-4调控系统对淋巴管生成和乳腺癌淋巴道转移的影响。方法： 体外培养胎牛胸导管内皮细胞，观察VEGF-C和抗Flt-4抗体对淋巴管内皮细胞增殖的影响；设计合成VEGF-C反义脱氧寡核苷酸（ASODN），体外实验观察其对VEGF-C基因表达的影响；建立乳腺癌裸鼠原位移植瘤模型并观察ASODN对肿瘤淋巴管生成及肿瘤生长转移的影响。结果： 加入前列腺癌细胞PC3(高表达VEGF-C)上清后，淋巴管内皮细胞增殖活跃；加入抗Flt-4抗体后各时段细胞计数均明显少于其它各组。体外实验RT-PCR及 Western blotting显示ASODN作用的MCF-7细胞组VEGF-C mRNA及蛋白表达均低于对照组。体内RT-PCR检测表明ASODN组移植瘤VEGF-C mRNA表达明显受抑制；5-Nase-ALPase双重酶组织化学法结果显示ASODN组移植瘤的淋巴管生成明显减少；ASODN组肿瘤生长速度较对照组缓慢，且肿瘤体积、淋巴结转移明显低于对照组。结论： VEGF-C/Flt-4调控系统与乳腺癌组织的淋巴管生成及肿瘤的淋巴道转移密切相关。阻断淋巴管内皮细胞Flt-4表达，可在一定程度上抑制肿瘤细胞诱导的淋巴管内皮细胞增殖；ASODN通过下调乳腺癌VEGF-C的表达，减少肿瘤淋巴管的生成及淋巴结转移。     

乳腺癌层粘连蛋白受体表达与微血管密度和转移及复发的关系

目的 :探讨乳腺癌层粘连蛋白受体 (LN R)与间质微血管密度和肿瘤转移、复发的关系及意义。方法 :应用免疫组化LSAB法检测 73例乳腺癌LN R、FⅧAg的表达 ,并对 2 8例病人进行随访分析。结果 :LN R的表达程度与乳腺癌淋巴结转移存在正相关 (P <0 0 0 0 1) ,与病理分级无关 (P >0 0 5 ) ,与乳腺癌间质微血管密度 (MVD)存在正相关 (P <0 0 0 0 1)。淋巴结转移阳性组其MVD值明显高于阴性组 (P <0 0 0 1)。随访 2 8例病人中 ,复发组的LN R表达增强比率及MVD值明显高于未复发组 (P <0 0 5 )。结论 :LN R的表达程度及MVD值均可作为判断乳腺癌病人转移及复发的独立指标     

VEGF、ANG-1、ANG-2、TSP-1的表达与胆管细胞性肝癌血管生成和侵润转移的关系

目的：研究血管内皮生长因子（VEGF）、血管生成素-1（ANG-1）、血管生成素-2（ANG-2）、血小板反应蛋白-1（TSP-1）的表达与胆管细胞性肝癌(CCC)血管生成和侵润转移的关系。方法： 对33例手术切除的CCC标本进行CD34、VEGF、 ANG-1、 ANG-2 和TSP-1的免疫组化染色，研究VEGF、ANG-1、ANG-2、TSP-1的表达与胆管细胞性肝癌血管生成和肿瘤门静脉侵犯、肝内转移、淋巴结转移以及肿瘤分化水平之间的关系。 结果： 本组CCC的微血管密度（MVD）为(87.2±52.6)/mm2，VEGF、ANG-1、ANG-2 和TSP-1的阳性率分别为75.6%、36.0%、57.6%和45.5%。VEGF和ANG-2的阳性表达与高MVD相关，TSP-1则与MVD负相关（P<0.01,P<0.05,P<0.01）。阳性TSP-1与肝内转移正相关（46.7% vs 5.6%，P<0.05）。结论： CCC瘤内的血管新生活跃，VEGF和ANG-2的阳性表达与CCC血管生成正相关，TSP-1则与其负相关，TSP-1的阳性表达还与肝内转移相关，VEGF、ANG-1、ANG-2的表达与肿瘤的侵润转移未见显著相关。     

The significance of c-erbB-2 overexpression and p53 expression in patients with axillary lymph node--negative breast cancer: a tissue microarray study

We conducted this study to examine whether the expression of c-erbB-2 and p53 is the prognostic indicator for patients with early-stage breast cancer in which axillary lymph node metastasis is absent. We examined 326 patients with early-stage breast cancer in which axillary lymph node metastasis is absent. Tissue microarrays were constructed. Following this, immunohistochemical staining was done for estrogen receptor (ER), progesterone receptor (PR), c-erbB2, and p53. The results were as follows: (1) expression of c-erbB-2 was correlated with other clinicopathologic factors (eg, patient's age, presence of menopause, tumor size, histologic and nuclear grade, and presence of hormone receptors such as ER and PR); and (2) expression of p53 was correlated with survival rate, patient's age, presence of menopause, and tumor size. However, these results were not statistically significant. In conclusion, our results indicate that expression of c-erbB-2 and p53 did not have any prognostic value in patients with early-stage breast cancer in which axillary lymph node metastasis is absent.     

多项生物指标在乳腺癌中的表达及其关系与预后

目的 探讨多项生物学指标c-erbB-2,PCNA,P53,EGFR,nm23,ER,PR在乳腺癌中的表达与临床乳腺癌预后的关系及这些指标间的相互关系。方法 应用免疫组织化学方法对87例乳腺癌组织切片c-erbB-2,PCNA,P53,EGFR,nm23,ER,PR的表达进行检测。结果 乳腺癌远处转移率与腋淋巴结转移数目有关，c-erbB-2阳性率与肿瘤大小，临床分期正相关，PCNA阳性率与肿瘤大小，腋淋巴结转移数目正相关，P53与肿瘤大小正相关，c-erbB-2与PCNA，P53，EGFR，PR相关，P53与PR相关，ER与PR相关。结论 肿瘤大小，腋淋巴结转移数目，临床分期，c-erbB-2,PCNA,P53,PR等多指标联合应用有助于提高对乳腺癌预后评价的正确性。     

Lymphatic Vessel Density and Vascular Endothelial Growth Factor Expression in Squamous Cell Carcinomas of Lip and Oral Cavity: A Clinicopathological Analysis with Immunohistochemistry Using Antibodies to D2-40, VEGF-C and VEGF-D

Background

Squamous cell carcinoma (SCC) of the oral region often metastasizes to the cervical lymph nodes. To investigate whether the risk of cervical lymph node metastasis are predictable through lymphatic vessel density (LVD) and vascular endothelial growth factor (VEGF) expression, we assessed the relationship between LVD and clinicopathological parameters, and VEGF expression in SCC of the oral region.

Methods

The subjects were 109 patients with SCC of the oral region including the lip. Clinicopathological parameters examined for the association with LVD in a peritumoral hot spot were lymph node metastasis, histological grade and disease stage. The association with VEGF expression was similarly studied. LVD was detected by immunohistochemistry using D2-40.

Results

LVD was significantly higher in lip cancer than in other oral tumors (P < 0.0001), while there were no significant differences of LVD among other cancers of the oral cavity. LVD tended to decrease with disease progression, increase of tumor size and increase of metastatic lymph node size. Eighty-four of 109 tumors were positive for VEGF-C or D. VEGF-C-positive tumor lesions were also positive for VEGF-D. Significantly higher levels of VEGF-C and D expressions were associated with large size of lymph node metastases (P = 0.02).

Conclusion

SCC of the oral region including the lip that produces VEGF-C and D is significantly more likely to cause cervical lymph node metastasis. LVD in a peritumoral hot spot does not directly indicate the risk of cervical lymph node metastasis, but instead may reflect lymphangiogenesis due to VEGF together with loss of lymphatic vessels through tumor growth and progression.     

血管内皮生长因子C和nm23表达与微血管密度及宫颈癌转移的关系

目的探讨血管内皮生长因子-C(vascular endothelial growth factor-C,VEGF-C)和nm23(non-metastasis23)表达与微血管密度(microvessel density,MVD)及宫颈癌转移的关系。方法采用免疫组化SABC法检测16例正常宫颈上皮及68例宫颈癌中VEGF-C、nm23的表达。并用CD105标记新生血管内皮细胞,计算肿瘤内MVD。结果VEGF-C在宫颈癌组织中的阳性表达率显著高于正常组织(P<0.01),淋巴结转移组VEGF-C阳性率高于无淋巴结转移组(P<0.05),VEGF-C表达强度与MVD呈正相关(P<0.05)。nm23蛋白表达率显著低于正常组织(P<0.01),淋巴结转移组nm23阳性率低于无淋巴结转移组(P<0.05)。nm23在宫颈癌中的表达与VEGF-C呈负相关(P<0.01)。结论宫颈癌组织VEGF-C呈高表达、nm23呈低表达,可能在宫颈癌血管生长、肿瘤浸润转移过程中起重要作用。检测宫颈癌中VEGF-C、nm23表达对进一步了解宫颈癌局部血管生成情况及判断预后有一定的应用价值。     

Vascular endothelial growth factor-C expression in human gallbladder cancer and its relationship to lymph node metastasis

Lymph node metastasis is a major prognostic factor in human cancer. Vascular endothelial growth factor C (VEGF-C) is a lymphangiogenic polypeptide that has been implicated in several human solid tumors. However, the clinical significance of VEGF-C has remained unknown in gallbladder carcinoma. Paraffin-embedded tumor specimens of 52 surgically resected gallbladder cancers were immunohistochemically stained for VEGF-C, VEGF, and CD34. The correlations among VEGF-C expression, VEGF expression, microvessel density (MVD), clinicopathologic features, and clinical outcomes were statistically analyzed. Thirty-three (63%) of the 52 gallbladder cancers were highly positive for VEGF-C protein by immunohistochemistry. VEGF-C expression was significantly correlated with lymphatic vessel involvement, lymph node metastasis, and worse outcomes after operation (p<0.001, p<0.001, p<0.001, respectively), but not with MVD. By the Cox regression model, lymphatic vessel involvement emerged as an independent prognostic parameter. These results suggest that VEGF-C may play a role in tumor progression via lymphangiogenesis and lymph node metastasis in human gallbladder cancer.     

Lymphatic vessel density in pulmonary adenocarcinoma immunohistochemically evaluated with anti-podoplanin or anti-D2-40 antibody is correlated with lymphatic invasion or lymph node metastases

In lung cancers, lymph node metastasis of cancer cells is one of the most important prognostic factors, and lymphatic vessel invasion (LVI) is very important in the stage preceding lymph node metastases. Recently, it has been reported that lymphatic vessel density (LVD) is associated with lymph node metastasis. The aim of the present study was to evaluate the relationship between LVD and LVI based on the immunohistochemical expression of podoplanin or D2-40, which are new specific markers for lymphatic endothelium. Using 76 cases of pulmonary adenocarcinoma, the relationship between LVD and LVI, lymph node metastases, vascular endothelial growth factor C (VEGF-C), VEGF-D or hepatocyte growth factor (HGF) expression was investigated. LVD was significantly associated with LVI, lymph node metastases and VEGF-D expression. LVI was also associated with lymph node metastases, histological subtype, VEGF-C or VEGF-D expression. High LVD, induced by VEGF-C or VEGF-D expression of cancer cells, is a good indicator of lymphatic metastases and LVI in pulmonary adenocarcinoma.     

Hypoxia-induced factor-1 alpha upregulates vascular endothelial growth factor C to promote lymphangiogenesis and angiogenesis in breast cancer patients

Hypoxia-induced factor-1 alpha （HIF-1α） affects many effector molecules and regulates tumor lymphangio- genesis and angiogenesis during hypoxia. The aim of this study was to investigate the role of HIF-1α in the regu- lation of vascular endothelial growth factor C （VEGF-C） expression and its effect on lymphangiogenesis and an- giogenesis in breast cancer. Lymphatic vessel density （LVD）, microvessel density （MVD） and the expressions of HIF-1α and VEGF-C proteins were evaluated by immunohistochemistry in 75 breast cancer samples. There was a significant correlation between HIF-1α and VEGF-C （P = 0.014, r = 0.273, Spearman＇s coefficient of correlation）. HIF-1α and VEGF-C overexpression was significantly correlated with higher LVD （P = 0.003 and P = 0.017, re- spectively）, regional lymph nodal involvement （P = 0.002 and P = 0.004, respectively） and advanced tumor, node, metastasis （TNM） classification （P = 0.001 and P = 0.01, respectively）. Higher MVD was observed in the group expressing higher levels of HIF-1α and VEGF-C （P = 0.033 and P = 0.037, respectively）. Univariate analysis showed shorter survival time in patients expressing higher levels of HIF-1α and VEGF-C. HIF-1α was also found to be an independent prognostic factor of overall survival in multivariate analysis. The results suggest that HIF-1α may affect VEGF-C expression, thus acting as a crucial regulator of lymphangiogenesis and angiogenesis in breast cancer. This study highlights promising potential of HIF- 1α as a therapeutic target against tumor lymph node me- tastasis.     

Smad4的表达与卵巢癌淋巴管生成及淋巴结转移之间的关系

目的观察Smad4和血管内皮生长因子(VEGF)-C在卵巢癌组织内的表达情况,分析Smad4和VEGF-C的表达与卵巢癌淋巴管生成及淋巴结转移之间的关系。方法取卵巢癌病例60例,其中,淋巴结转移组36例,无淋巴结转移组24例。应用免疫组化法和Westernblot技术观察Smad4和VEGF-C在卵巢癌组织内的表达。以D2-40作为淋巴管内皮特异性标记物,检测卵巢癌组织内淋巴管生成情况。结果 Smad4表达于卵巢癌细胞胞浆和胞核内,其在无淋巴结转移组的表达率明显高于其在有淋巴结转移组的表达率。Smad4表达阳性组的淋巴管数密度(LVD)明显低于Smad4表达阴性组的LVD。VEGF-C主要表达于卵巢癌细胞胞浆内,其在淋巴结转移组的表达率明显高于其在无淋巴结转移组的表达率。Smad4的表达与VEGF-C的表达呈显著的负相关性。Western blot检测结果表明,VEGF-C蛋白在有淋巴结转移卵巢癌组织中的表达量高于其在无淋巴结转移卵巢癌组织内的表达量,而Smad4在有淋巴结转移卵巢癌组织内的表达量明显低于其在无淋巴结转移组的表达量。结论 Smad4与VEGF-C的表达呈负相关,Smad4可能通过调节VEGF-C蛋白的表达而抑制卵巢癌淋巴管生成和淋巴道转移。     

Expression of vascular endothelial growth factor-C and its receptor in invasive micropapillary carcinoma of the breast

Invasion to lymphatic vessels and metastasis to lymph nodes are frequent complications in invasive micropapillary carcinoma (IMPC) of human breast cancer. Vascular endothelial growth factor-C (VEGF-C) and its receptor, VEGFR-3 have been implicated as the important factors in the formation of lymphatic vessels and recent experimental evidence strongly suggests that lymphangiogenesis in tumor promotes lymphatic metastasis. To clarify the mechanism of its occurrence, the expression of VEGF-C, VEGFR-3 and lymphatic vessel density (LVD) was examined in 40 cases of IMPC (pure and mixed type) and in 40 cases of pseudo-IMPC. Cytoplasmic expression of VEGF-C and VEGFR-3 were more frequent in tumor cells of IMPC compared to those of pseudo-IMPC. A significant positive correlation was found between the expression of VEGF-C and VEGFR-3 in both IMPC and pseudo-IMPC. The expression of VEGF-C was also significantly associated with higher peritumoral LVD, lymphatic invasion and number of lymph node metastasis in IMPC. These findings suggest that VEGF-C promotes the proliferation of peritumoral lymphatic vessels and that lymphatic invasion and metastasis to lymph nodes are frequently induced in IMPC of breast.