T淋巴细胞免疫功能重建对小鼠肝癌转移的影响

目的研究T淋巴细胞免疫功能重建对小鼠肝癌转移程度的影响,进一步探讨免疫对肝癌转移的影响因素,寻找肝癌转移的免疫干预手段。方法建立T淋巴细胞免疫功能重建的模型。将裸鼠分成高、低转移的T淋巴细胞免疫功能重建组和未重建组。取淋巴结通过病理切片观察转移程度。计算成瘤时间和生长速度的差异,检测肿瘤细胞株组织相容性复合物(MHC)Ⅰ、Ⅱ表达。应用Th1—Th2-Th3基因芯片检测不同转移潜能小鼠体内T淋巴细胞相关基因的差异。结果进行T淋巴细胞免疫功能重建后裸鼠外周血CD 3、CD4、CD 8和CD4/CD 8比例均高于未重建组;高、低转移潜能的肝癌细胞在裸鼠体内平均成瘤时间为(7.7±0.6)d,正常Balb/c小鼠为(11.5±1.3)d。高、低转移潜能肝癌细胞的肿瘤生长速度具有明显的差异。免疫重建后的裸鼠肿瘤淋巴结远处转移显著降低(P<0.0 5)。高转移组肿瘤细胞的M H C Ⅰ表达明显低于低转移组(P<0.0 5)。高转移组T淋巴细胞Th1/Th2相关基因的表达低于低转移组。结论免疫功能重建可以在一定程度上影响小鼠肝癌的转移。肿瘤M H C分子表达的改变和机体淋巴细胞T h 1/T h 2 相关基因的表达降低可能是影响肝癌转移潜能的原因。     

小鼠口腔癌不同时期淋巴结和骨髓的VEGF表达情况及其微环境对淋巴道转移的影响

目的 探讨小鼠口腔癌不同时期淋巴结和骨髓的血管内皮生长因子(VEGF)表达情况,及其微环境对口腔癌淋巴道转移的影响.方法 选入40只6周龄雄性健康Balb/c小鼠,将其随机分为A、B、C、D、E组,每组8只.采用4-硝基喹啉-1-氧化物(4-NQO)饮水法构建小鼠口腔癌淋巴道转移模型.其中A组为正常小鼠;B组小鼠舌部黏...     

消化道恶性肿瘤淋巴管生成的研究进展

消化道恶性肿瘤的发病率日益增高,而肿瘤的淋巴道转移是导致患者最终死亡的重要因素之一．在某些恶性肿瘤中已经证实有淋巴管生成,被称为淋巴管生成因子的VEGF-C、 VEGF-D及其受体VEGFR-3能够诱导淋巴管的生成．恶性肿瘤中的淋巴管生成因子不但能够促进肿瘤淋巴管的生成,而且与淋巴道转移密切相关．近年来对于消化道恶性肿瘤的淋巴管生成以及淋巴道转移的研究正在开展,本文将对消化道恶性肿瘤淋巴管生成机制,淋巴管生成与肿瘤淋巴道转移的关系,最新研究进展以及此项研究的临床意义加以综述．     

淋巴管新生与肿瘤淋巴道转移的相关性

在实体恶性肿瘤中,淋巴道是肿瘤细胞转移的主要途径,淋巴管生成在肿瘤的淋巴转移过程中起重要作用。随着淋巴管内皮特异标记物和淋巴管生长因子的发现,肿瘤诱发淋巴管生成的分子机制和以抑制肿瘤淋巴管生成作为抗肿瘤转移靶点的研究引起临床关注。现将淋巴管生成于肿瘤淋巴道转移相关性的研究进展综述如下。     

血管内皮生长因子C与肺癌关系的研究进展

包括肺癌在内的恶性肿瘤是严重危害人民健康的疾病。近年，血管内皮生长因子（VEGF）家族与肿瘤关系的研究成为热点。血管内皮生长因子C（VEGF—C）是VEGF家族的新成员，作为第一个被发现的促淋巴管生成因子，由于其强大、特异的促淋巴管生成作用，在与肿瘤、特别是肿瘤淋巴转移的相关研究中备受关注。现就近年VEGF—C与肺癌关系的研究进展作一综述。     

大肠癌血管内皮生长因子的表达及其意义

目的：探讨血管内皮生长因子（VEGF）mRNA在大肠癌和癌周组织中的表达及其与临床特征之间的关系,为在分子水平干预肿瘤血管形成,预防大肠癌复发和转移打下基础。方法：采用逆转录－聚合酶链反应（RT－PCR）检测方法,对68例大肠癌手术标本癌和癌周组织中VEGF mRNA的表达水平进行了相对定量研究。结果：肿瘤组织中VEGF mRNA的表达率为67．6％（46／68）,癌周组织表达率为32．4％（22／68）;肿瘤组织中VEGF mRNA表达水平在浆膜浸润组,淋巴结转移,远处转移和Dukes D期组分别显著高于未侵及浆膜组,无淋巴结转移组,无远处转移和Dukes A,B,C期组;肿瘤组织中VEGF mRNA表达水平在肿瘤直径大的大肠癌组（＞5cm)与大小肠癌组（≤5cm)以及不同组织学分组组之间相比差异无显著性。结论：VEGF在大肠癌浸润和转移过程中发挥重要作用,肿瘤血管形成与大肠癌浸润和转移关系密切。     

肺腺癌组织中VEGF-D的表达变化及意义

目的观察肺腺癌组织中血管内皮生长因子D（VEGF—D）的表达,探讨其意义。方法分别采用RT-PCR法、免疫组化法检测48例肺腺癌组织中的VEGF—D mRNA和VEGF—D、微淋巴管密度（MLVD）、微血管密度（MVD）。结果VEGF—D mRNA在肺腺癌组织中的表达高于正常肺组织（P〈0．01）,VEGF—D蛋白阳性率肿瘤周边显著高于肿瘤中心（P〈0．01）;其表达与肿瘤分化、MVD无关,与TNM分期、MLVD、淋巴结转移有关（P均〈0．05）。结论肺腺癌组织中VEGF—D高表达,与淋巴管的生成及转移有关。     

血管内皮生长因子-C表达与非小细胞肺癌淋巴管生成和淋巴结转移的相关性

目的观察血管内皮生长因子(VEGF)-C在非小细胞肺癌(NSCLC)组织中央部位、侵犯边缘和癌周肺组织的表达情况,分析VEGFC与淋巴结转移、淋巴管侵犯和淋巴管生成的相关性。方法选择人NSCLC新鲜组织样本96例,采用免疫组织化学法和实时定量RT-PCR(QRTPCR)法检测NSCLC组织中央部位、侵犯边缘和癌周肺组织的VEGF-C蛋白及mRNA表达情况,用D2-40标记淋巴管检测NSCLC组织中的淋巴管密度。结果 QRT-PCR显示,VEGF-C mRNA在肿瘤边缘部位的表达明显高于在肿瘤中央部位的表达,而且VEGF-C mRNA的表达亦高于其在周围肺组织的表达。免疫组织化学结果显示了同样的表达不均一性。肿瘤边缘VEGF-C高表达组淋巴结转移的发生率明显高于低表达组,而且VEGF-C高表达组淋巴管侵犯的发生率亦明显增高。肿瘤边缘高淋巴管密度组的VEGF-C mRNA的表达也明显高于低密度组。结论肿瘤侵犯边缘的VEGFC高表达与淋巴结转移和淋巴管侵犯密切相关,参与了NSCLC淋巴管生成和淋巴结转移过程。     

膜-细胞骨架联接分子ezrin对肝癌细胞系生长和侵袭能力的影响

目的探讨膜-细胞骨架联接蛋白ezrin在肝细胞肝癌生长和转移过程中的作用。方法分别应用免疫荧光、逆转录聚合酶链反应（RT—PCR）和Western blot检测ezrin和骨架蛋白在不同转移潜能肝癌细胞系中的表达。选取高转移潜能SF7721（SMMC-7721经转基因后稳定表达肝细胞生长因子，从而获得高转移潜能的细胞系）和MHCC97-H细胞系为研究对象。通过RNA干扰技术下调SF7721和MHCC97-H细胞系中ezrin蛋白的表达，观察其运动和侵袭能力的变化：通过四甲基偶氮唑盐检测细胞增殖能力变化；电子显微镜观察细胞伪足；Transwell检测细胞的运动侵袭能力。结果免疫荧光显示ezrin和骨架蛋白表达于细胞质，且双色荧光证实两者存在共表达，高转移潜能细胞系SF7721。MHCC—I、MHCC97-Hezrin和骨架蛋白的表达明显高于低转移潜能细胞系SMMC-7721、Hep3B、HepG2细胞（x^2=13．277，P=0．010；x^2=21．815，P〈0．01）。D-肌动蛋白在高低转移潜能细胞系的表达差异无统计学意义。通过RNA干扰技术抑制ezrin蛋白表达后。SF7721和MHHC97-H的细胞的增殖和侵袭能力均显著下降。结论ezrin和骨架蛋白的过表达与肝癌的转移潜能相关，通过下调ezrin的表达可明显抑制肝癌细胞系SMMC-7721和MHCC97-H细胞的增殖和运动侵袭能力。     

环氧合酶-2和血管内皮生长因子-C与胃癌淋巴管转移

目的　研究环氧合酶 2 (COX 2 )和血管内皮生长因子 (VEGF) C在胃癌组织中的表达及相关性 ,探讨二者在胃癌淋巴管生成和转移中的作用。方法　采用免疫组化方法和逆转录 PCR技术 ,分别检测了 6 4例胃癌石蜡组织中COX 2和VEGF C的表达及其中 2 2例胃癌新鲜组织中二者mRNA的表达。结果　 2 2例胃癌新鲜组织中COX 2和VEGF CmRNA表达阳性率分别为 82 %和73% ,其表达均高于相应的癌旁正常组织 (P <0 0 0 1) ,与 6 4例胃癌石蜡标本COX 2和VEGF C的表达结果较一致。且COX 2和VEGF C表达之间存在明显关联性 (P <0 0 5 )。二者在癌组织中的高表达与肿瘤浸润深度、淋巴结转移等密切相关 (P <0 0 5 )。结论　胃癌组织中有COX 2和VEGF C的高表达 ,而COX 2可能参与VEGF C淋巴管生成通路 ,它们的表达可能在胃癌淋巴管浸润和转移中发挥重要作用     

VEGFR-3信号途径相关蛋白在胃癌中的表达及意义

目的研究血管内皮生长因子受体（VEGFR）-3信号途径相关蛋白C、D（VEGF—C、VEGF—D）在胃癌中的表达并探讨其与淋巴结转移的关系。方法80例胃癌手术病例分为淋巴结阳性组（n=48）和淋巴结阴性组（n=32）,另设胃溃疡病例为对照组（n=10）。采用ELISA技术进行血清VEGF—C和VEGF-D水平检测,然后应用免疫组织化学EnVisionTM两步法检测胃溃疡组织和胃癌标本VEGF—C、VEGF-D表达。结果胃癌患者血清VEGF-C和VEGF—D水平明显高于对照组（P〈0．05）,胃癌患者血清VEGF-C水平与淋巴结转移有关（P〈0．05）;胃癌组织中VEGF—C、VEGF—D阳性表达率均高于对照组（P〈0．05）,伴淋巴结转移的胃癌组织VEGF-C阳性表达率较无淋巴结转移者更高（P〈0．01）。结论血清VEGF—C可作为胃癌的标记物,对术前判定淋巴结转移具有一定的临床价值。     

食管鳞癌组织中VEGF-D表达及其与淋巴管密度和淋巴转移的关系

目的 研究食管鳞状细胞癌(ESCC)组织中血管内皮细胞生长因子D(VEGF-D)的表达情况及其与淋巴管密度(LVD)的关系,并探讨其与食管癌淋巴结转移的关系.方法 免疫组化SABC法检测55例手术切除的食管鳞状细胞癌组织、癌旁组织、正常组织中淋巴管密度和VEGF-D的表达水平,对结果进行统计分析.结果 (1)食管癌组织...     

大肠癌组织中生长抑素和血管内皮生长因子-C的表达及其临床意义

目的探讨大肠癌组织中生长抑素（SS）和血管内皮生长因子-C（VEGF-C）的表达及其临床意义。方法采用免疫组化法检测60例大肠癌组织及20例正常大肠黏膜组织中SS蛋白和VEGF-C蛋白的表达，采用VEGF-C受体FLT-4阳性脉管标记淋巴管计数，分析SS与大肠癌淋巴管生成、转移的关系。结果SS蛋白表达阳性率在大肠癌组及正常大肠黏膜组分别为37．7％和65％，VEGF-C在癌及正常组阳性表达率分别为60％和30％，差别均有显著性；SS表达与肿瘤分化程度、淋巴结转移、淋巴管侵犯及远处转移密切相关，与浆面膜受累无关；VEGF-C表达与肿瘤淋巴结转移，淋巴管侵犯及远处转移密切相关，而与肿瘤分化和浆面膜受累无关；大肠癌组织中SS和VEGF-C蛋白表达呈显著负相关。结论SS可能通过对VEGF-C／FLT-4信号通路的阻滞而抑制大肠癌淋巴管生成及转移。     

Vascular endothelial growth factor-C (VEGF-C) is a more specific risk factor for lymph node metastasis than VEGF-D in submucosal colorectal cancer

BACKGROUND/AIMS: It is useful to decide whether lymphatic involvement or lymph node metastasis exists before polypectomy or operation in submucosal colorectal cancer. Whether vascular endothelial growth factor-C (VEGF-C) or VEGF-D could predict lymph node metastasis and lymphatic involvement is uncertain. METHODOLOGY: Expression of the VEGF-C and VEGF-D in human submucosal colorectal cancers was investigated in paraffin-embedded stepwise sections by means of immunohistochemistry, and the correlation between immunohistochemical expression pattern and clinicopathological features was also evaluated. RESULTS: The results showed that VEGF-C overexpression correlated with lymphatic involvement (P = 0.01) and lymph node metastasis (P = 0.02), but VEGF-D overexpression did not correlate significantly. In multivariate analysis lymphatic invasion was the predictive factor (P = 0.0129), but VEGF-C positivity was not predictive (P = 0.3437). CONCLUSIONS: These results may suggest that VEGF-C is a more specific risk factor for lymph node metastasis than VEGF-D in submucosal colorectal cancer.     

胃癌淋巴结转移与VEGF—D、VEGFR-3、LVD的关系

目的探讨胃癌淋巴结转移与血管内皮生长因子D（VEGF—D）、血管内皮生长因子受体3（VEGFR-3）、淋巴管密度（LVD）表达间的关系。方法对59例胃癌术后标本采用免疫组织化学sP法检测VEGF—D表达,用VEGFR-3抗体免疫组织化学sP法标记淋巴管,检测肿瘤组织及正常组织LVD。结果VEGF—D、VEGFR-3在癌组织中表达率分别为59．32％、67．80％,明显高于癌旁不典型增生组织及正常胃黏膜组织（P〈0．05）。癌周组织的LVD表达为（21．29±8．21）,明显高于周边正常组织（P〈0．05）。VEGF—D表达阳性组35例与阴性组24例的LVD分别为（23．15±7．58）和（11．93±5．31）,其差异具有统计学意义（P〈0．05）。VEGF—D、LVD在癌组织中的表达与淋巴结直径、淋巴结分期、浸润深度、TNM分期及分化程度有关（P〈0．05）。结论VEGF．D在胃癌组织中高表达,并与肿瘤的淋巴管形成及淋巴结转移密切相关。     

Expression of vascular endothelial growth factor-C and the relationship between lymphangiogenesis and lymphatic metastasis in colorectal cancer

AIM: To investigate the expression of vascular endothelial growth factor-C (VEGF-C) and the relationship between VEGF-C and lymphangiogenesis, lymph node metastasis in colorectal cancer. METHODS: Fifty six cases of colorectal cancer were selected randomly. Expression of VEGF-C was detected by immunohistochemistry, and lymphatic vessels were stained by enzyme histochemical method. RESULTS: VEGF-C expression was found in 66.7% (37/56) patients. In VEGF-C positive and negative patients, the lymphatic vessel density was 25.16+/-7.52 and 17.14+/-7.22, respectively (P<0.05). The rate of lymph node metastasis in VEGF-C positive patients (81.1%) was significantly higher than that in the negative group (42.1%). CONCLUSION: VEGF-C expression may induce lymphangiogenesis in colorectal cancer, as a result, tumor cells can entry the lymphatic vessels easily. VEGF-C may serve as a useful prognotic factor in colorectal carcinoma.     

Vascular endothelial growth factors C and D and their VEGFR-2 and 3 receptors in blood and lymphatic vessels in healthy and arthritic synovium

OBJECTIVE: To localize vascular endothelial growth factor C (VEGF-C) and VEGF-D in synovial specimens in relation to their VEGFR-2 and VEGFR-3 receptors in blood and lymphatic vessels. METHODS: Immunohistochemical staining and messenger RNA analysis from control and arthritic synovial membrane specimens. RESULTS: Quantitative RT-PCR disclosed that VEGF-C mRNA copy numbers were higher than VEGF-D mRNA copy numbers in the rheumatoid arthritis (RA), osteoarthritis, and control patient groups studied (p < 0.01). Immunohistochemical staining localized VEGF-C to synovial lining cell layer, pericytes, and smooth muscle cells of blood vessels. The number of VEGF-C positive cells was increased in the synovial lining of ankylosing spondylitis (AS) and RA compared to control synovium. However, in contrast to control synovial lining, little if any VEGF-D was detected in AS or RA synovial lining. VEGFR-2 expressing stromal blood vessels, also positive for the vascular endothelial marker PAL-E and the basement membrane marker laminin, were more abundant in RA and AS than in controls. Interestingly, the lymphatic endothelial receptor VEGFR-3 was also expressed in most synovial vessels, especially in the sublining capillaries and venules. CONCLUSION: VEGF-C is strongly expressed in the hypertrophic synovial lining of arthritic joints, whereas VEGF-D expression is very low in AS and RA. The expression of VEGF-C and VEGF-D in pericytes and smooth muscle cells suggests that these factors may have a role in maintaining vascular homeostasis. The VEGF receptors VEGFR-2 and VEGFR-3 are present in most of the sublining blood vessels. The expression of the lymphatic marker VEGFR-3 in the sublining blood vessels may relate to fluid filtration and/or fenestrations. The relatively few lymphatic vessels along with increased vascular permeability in RA may contribute to the development of tissue edema and joint stiffness.     

KAI1 gene expression in colonic carcinoma and its clinical significances

AIM: To investigate KAI1 gene expression in the progression of human colonic carcinoma and its clinical significances. METHODS: KAI1 expression was detected by in situ hybridization and immunohistochemistry in the 4 established cell lines of colorectal carcinoma with different metastatic potentials, and in 80 specimens of colonic carcinoma, 21 colonic carcinoma specimens with lymphatic metastasis and 20 controls of normal colonic mucosa. RESULTS: The expressions of KAI1 in HT29 and SW480 cell lines were higher than those in LoVo and SW620. The expression of KAI1 gene was significantly higher in colorectal carcinoma compared with normal colonic mucosa and lymphatic metastasis (chi(2)=46.838, P<0.01). The expression of KAI1 gene had no relationship with histological grade. The KAI1 expressions in Dukes A and B carcinoma were higher at both mRNA and protein levels compared to Dukes C carcinoma (chi(2)=16.061, P<0.05). The expression of KAI1 in colonic carcinoma specimens with lymphatic metastasis was almost lost. The results of in situ hybridization were in concordance with immunohistochemistry. CONCLUSION: KAI1 is highly related to the metastasis of colonic carcinoma and may be a useful indicator of metastasis in colonic carcinoma.     

肿瘤抗淋巴管生成研究进展

肿瘤转移是癌症患者的主要死因之一,淋巴管转移是肿瘤转移的重要途径,研究发现VEGF-C/VEGF-D/VEGFR-3与肿瘤淋巴管生成、肿瘤转移、肿瘤预后密切相关.大量的研究证实VEGF-C/VEGF-D/VEGFR-3信号传导轴在调节肿瘤淋巴管生成中起主导作用,临床病理研究也显示VEGF-C/VEGF-D/VEGFR-...     

结肠癌患者外周血VEGF-C和CK20 mRNA的表达与淋巴转移的关系

目的探讨结肠癌患者外周血VEGF-C mRNA和CK20 mRNA的表达与淋巴转移的关系。方法应用RT-PCR检测80例结肠癌患者、30例结肠良性肿瘤患者、30例健康人外周血VEGF-C mRNA和CK20 mRNA的表达。结果 VEGF-C mRNA在结肠癌患者外周血的阳性表达率为52.5%,在结肠良性肿瘤、健康人的表达率分别为10.0%、0,其中有淋巴结转移的结肠癌患者的阳性表达明显高于无淋巴结转移的阳性表达(P<0.05)。CK20 mRNA在结肠癌患者外周血的阳性表达率为60.0%,在结肠良性肿瘤、健康人的表达率均为0,其中有淋巴结转移的结肠癌患者的阳性表达明显高于无淋巴结转移的阳性表达(P<0.05)。结论结肠癌患者外周血VEGF-C mRNA和CK20 mRNA的表达与淋巴转移相关。