非小细胞肺癌VEGF-C、VEGF-D、VEGFR-3表达与淋巴管生成和淋巴转移关系的研究

Investigation on the relationship between VEGF-C,VEGF-D,VEGFR-3 and lymphangio-genesis and lymph node metastasis in non-small cell lung cancer

目的:探讨血管内皮生长因子(VEGF)-C、-D及其受体3(VEGFR-3)在非小细胞肺癌(NSCLC)中的表达规律、与微淋巴管密度(MLVD)、淋巴转移之间的关系及其在NSCLC发生发展、预后中的意义。方法:以40例经病理确诊的NSCLC组织为实验组,以11例肺良性病变组织为对照组,采用免疫组织化学染色法对上述组织中VEGF-C、VEGF-D、VEGFR-3、淋巴管内皮透明质酸受体-1(LYVE-1)、同源异型盒转录因子(Prox-1)蛋白的表达进行分析,并计数微淋巴管密度。结果:①NSCLC组织中VEGF-C、VEGF-D及VEGFR-3蛋白的阳性率分别为77.50%(31/40例)、67.50%(27/40例)、62.50%(25/40例),明显高于肿瘤周边组织(距肿瘤边缘100μm区域内)及肺良性病变组织(P<0.01)。②VEGF-C与VEGFR-3(r=0.409,P=0.005)、VEGF-D与VEGFR-3蛋白表达(r=0.492,P=0.000)均存在相关性;而VEGF-C与VEGF-D蛋白表达无相关性(r=0.256,P=0.093)。③VEGF-C、VEGF-D及VEGFR-3蛋白的表达与NSCLC患者的性别、年龄、肿瘤的大小、组织学类型、分化程度无关,但与肿瘤的淋巴结转移(P<0.05)、PTNM分期(P<0.05)显著相关。④NSCLC肿瘤中心组织中LYVE-1及Prox-1标记的MLVD分别为4.22±1.25、1.99±1.49,明显低于肺良性病变组织(P=0.000),而NSCLC肿瘤周边部位的MLVD显著高于肿瘤中心部位及肺良性病变组织(P=0.000)。VEGF-C、VEGF-D、VEGFR-3表达阳性的组织中,MLVD显著高于阴性组织(P<0.05)。MLVD与肿瘤的淋巴结转移(P=0.000)、PTNM分期(P=0.000)显著相关。结论:淋巴管生成因子VEGF-C、VEGF-D及其受体VEGFR-3蛋白在NSCLC中表达显著增高,并且通过VEGF-C、VEGF-D/VEGFR-3信号通路诱导淋巴管内皮细胞新生和淋巴管生成,从而促进淋巴结转移和肿瘤生长;VEGF-C、VEGF-D及其受体VEGFR-3可能成为检测NSCLC淋巴转移和评估预后的重要分子指标;淋巴管内皮细胞特异性标志物LYVE-1、Prox-1可以较严格地区分血管和淋巴管内皮,相对精确地评价肿瘤的脉管系统。

Objective：To investigate the expression of vascular endothelial growth factor （VEGF）-C,VEGF-D and their receptor-3 （VEGFR-3） in patients with non-small cell lung cancer （NSCLC）,and the relationship with lymphangiogenesis and lymph node metastasis and their clinicopathological significance. Methods：Forty specimens of the NSCLC and eleven benign pulmonary diseases were studied. The expressions of VEGF-C, VEGF-D, VEGFR-3, LVYVE-1 and Prox-1 protein in specimens of NSCLC and benign pulmonary diseases tissues were studied by immunohistochemical technique. Microlymphatic vessel density （MLVD） was counted. Results： ①Among 40 cases of NSCLC, the positive rates of VEGF-C, VEGF-D and VEGFR-3 were 77.50%（31/40）, 67.50%（27/40） and 62.50% （25/40） respectively,which were significantly higher than those of the paraneoplastic and pulmonary benign diseases tissues （P 〈 0.01）. ②Significant correlations between VEGF-C and VEGFR-3 （r=0.409,P=0.005）, VEGF-D and VEGFR-3 （r=0.492,P=0.000） expression were observed in NSCLC;while no correlation between VEGF-C and VEGF-D（r=0.256,P=0.093） was observed. ③The expressions of VEGF-C,VEGF-D and VEGFR-3 protein in NSCLC were not correlated with age,gender,site and dimension of lesion,types of histological and degree of differentiation,but correlated significantly with lymph node metastasis （P 〈 0.05） and PTNM stage （P 〈 0.05）. ④In the center of NSCLC cancerous tissues among 40 cases,the microlymphatic vessel densities marked by LYVE-1 and Prox-1 were 4.22± 1.25 and 1.99 ± 1.49 respectively,which were significantly lower than those of the pulmonary benign diseases tissues （P=0.000）. However, the MLVDs in cancerous invasive edge were significantly higher than those in the center of cancerous tissues and those in the pulmonary benign diseases tissues （P=0.000）. The MLVDs in the positive VEGF-C, VEGF-D and VEGFR-3 groups were significantly higher than those in the negative groups （P〈 0.05）. MLVDs were correlated significantly with lymph node metastasis （P=O.O00） and PTNM stage （P=0.000）. Conclusion：Non-small cell lung cancer ceils may secrete lymphangiogenetic growth factors VEGF-C,VEGF-D and their receptor VEGFR-3, which induce the growth of lymphatic vessel endothelium and lymphangiogenesis by VEGF-C, VEGF-D/VEGFR-3 signaling pathway, and further accelerate lymphatic metastasis of NSCLC. VEGF-C,VEGF-D and VEGFR-3 might be acted as molecular phenotypes of lymphangioghesis in NSCLC and important markers for evaluating lymphaticmetastasis and prognosis in patients with NSCLC. As specific markers of lymphatic vessel endotheliocyte,LYVE-1 and Prox-1 could identify blood vessels and lymphatic vessels strictly and evaluate vascular system of tumor fairly exactly.