血管内皮细胞生长因子与增殖细胞核抗原联合检测用于大肠癌临床预后判定

目的　探讨大肠癌血管内皮细胞生长因子 (VEGF)及增殖细胞核抗原 (PCNA)与大肠癌术后有无潜在性转移及复发的关系。方法　对 5 9例已获确诊的大肠癌石蜡标本作免疫组化SP法染色 ,检测其VEGF及PCNA的表达 ,并与 12例正常大肠组织和 16例大肠腺瘤进行比较。结果　大肠癌VEGF的表达明显高于大肠腺瘤 (P<0 .0 5 ) ;DukesA +B期大肠癌VEGF的表达与DukesC +D期比较 ,差异有显著性 (P<0 .0 5 ) ;术后复发组VEGF的表达明显高于无复发组 (P<0 .0 5 )。增殖活性表达提示 ,大肠癌分化程度越低 ,有淋巴结或肝转移时 ,其PCNA指数增高 ;术后有、无复发者之间 ,其PCNA指数差异有显著性 (P<0 .0 5 )。结论　大肠癌VEGF与PCNA的表达与肿瘤的浸润、转移密切相关。手术时虽然无明显转移灶 ,VEGF阳性及PCNA活性增强时仍可能有潜在的转移存在。     

大肠癌围手术期血清血管内皮生长因子表达与预后的关系

为探讨大肠癌围手术期患者血清血管内皮生长因子（VEGF）表达与预后的关系,采用酶链免疫吸附测定试剂盒检测8名健康体检者和40例大肠癌患者术前与术后1周血清VEGF的含量,并分析大肠癌患者术后VEGF水平下降程度对持续无瘤生存期的影响。结果显示,大肠癌患者术前VEGF水平为（497．68±128．36）pg／ml,术后1周为（368．56±98．72）Pg／ml,均明显高于健康体检者（224．54±68．23）pg／ml,q术前=8．951,q术后=4．720,P均〈0．01。大肠癌患者术后1周VEGF水平明显下降,q=7．329,P〈0．01。术后VEGF水平下降〈50％组持续无瘤生存期低于VEGF水平下降≥50％组,x2=8．903,P〈0．05。结果表明,VEGF在大肠癌患者血清中高表达,与患者预后显著相关,可作为判断大肠癌预后的有效指标。     

Profile of Plasma Angiogenic Factors Before and After Hepatectomy for Colorectal Cancer Liver Metastases

Background Circulating angiogenic factors in patients with colorectal cancer liver metastases may promote tumor growth and contribute to liver regeneration after partial hepatectomy. Methods We analyzed blood samples from 26 patients with colorectal cancer liver metastases before and after liver resection and used samples from 20 healthy controls as a reference. Plasma levels of vascular endothelial growth factor (VEGF), basic fibroblast growth factor (bFGF), epidermal growth factor (EGF), and hepatocyte growth factor (HGF) were measured, and levels were correlated with recurrence. Results The median preoperative levels of all four factors were significantly higher and more variable in colorectal cancer liver metastasis patients than in controls. HGF and bFGF levels increased significantly 3 days and 1 month after hepatectomy, respectively, and returned to near preoperative levels at 3 months. Postoperative VEGF and EGF levels remained relatively stably increased over 3 months. After a median follow-up of 19 months, 10 patients (42%) experienced recurrence. Higher preoperative VEGF and HGF levels correlated with subsequent recurrence (P = .018 and .021, respectively), and a preoperative adjusted total value of all four factors accurately identified patients at low, moderate, and high risk of recurrence (P = .034). Patients who experienced disease recurrence also had relatively higher bFGF levels 3 months after operation (P = .035). Conclusions Plasma angiogenic factors are increased in patients with colorectal cancer liver metastases and remain increased at least 3 months after partial hepatectomy. Measurement of certain factors before and after hepatic resection can predict recurrence. Targeted biological agents may counteract the tumor-promoting effects of these circulating factors on subclinical disease.     

乳腺癌患者血清和肿瘤组织VEGF表达与临床预后的关系

目的探讨乳腺癌患者血清和肿瘤组织中血管内皮细胞生长因子（VEGF）的表达以及与临床预后的关系。方法以44例乳腺癌患者、13例乳腺良性疾病患者和40例正常人为研究对象，分别采用酶联免疫吸附法（ELISA）检测其血清VEGF水平，采用免疫组化LSAB法检测其组织中VEGF、雌激素受体（ER）和原癌基因C-erbB-2蛋白的表达，并分析其与临床预后因素如淋巴结转移情况及临床分期的关系。结果乳腺癌组血清和组织中VEGF表达水平（113．88pg／ml，20723．99）均明显高于乳腺良性疾病组（55．79pg／ml，3594．74），P〈0．001，而乳腺良性疾病组与正常对照组（41．30pg／ml，-）比较差异无统计学意义（P〉0．05）；且血清和组织中VEGF的表达呈正相关（r=0．48，P〈0．01）。有淋巴结转移者血清和组织中VEGF的表达水平（129．60pg／ml，28506．82）明显高于无淋巴结转移者（80．80pg／ml，14656．73），P〈0．01；血清和组织中VEGF的表达与肿瘤的临床分期有关（P〈0．01），但与患者年龄和肿瘤大小无关（P〉0．05）。乳腺癌患者血清和组织中VEGF表达水平与组织中ER表达呈负相关（r=-0．45，P〈0．05），与C-erbB-2表达呈正相关（r=0．48，P〈0．01）。结论乳腺癌患者血清和肿瘤组织中VEGF表达呈正相关，且与其临床预后有关，可作为乳腺癌患者预后判断的重要参考指标之一。     

大肠癌组织VEGF-C表达与血管生成和细胞增殖关系的研究

目的：研究大肠癌组织血管内皮生长因子C（VEGF—C）表达与肿瘤血管生成及肿瘤细胞增殖的关系。方法：采用SP免疫组织化学方法，检测81例大肠癌VEGF-C、CD34和Ki-67的表达，并分析VEGF-C与微血管密度（MVD）、Ki-67增殖指数（Ki-67PI）的相关性。结果：81例大肠癌中，VEGF-C表达阳性45例（55．56％）；VEGFC表达阳性的肿瘤其MVD和Ki-67PI高于VEGF-C者（P〈0.05）。VEGF-C表达与MVD、Ki-67PI呈正相关（P〈0.05）。有转移组（包括淋巴结转移和远处转移）的VEGF-C表达明显高于无转移组，表达随肿瘤分期的升高而增强。结论：VEGF-C通过诱导新生血管形成，促进肿瘤细胞的增殖生长，影响着大肠癌的浸润和转移。     

Elevated Serum Vascular Endothelial Growth Factor and Basic Fibroblast Growth Factor Levels in Patients with Thymic Epithelial Neoplasms

Neovascularization, an essential event for the growth of solid tumors, is regulated by a number of angiogenic factors, among which vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF), are considered to exert potent angiogenic activity. In this study, we investigated whether serum VEGF and bFGF levels could be predictors of the development and extension of thymic epithelial neoplasms. The subjects of this study were 37 patients with thymoma, 6 with thymic carcinoma, and 23 healthy volunteers. Serum samples were collected before clinical treatment. Serum VEGF levels were significantly (P < 0.05) elevated in the patients with thymic carcinoma (1 080 ± 1 185 pg/ml) compared with those in the healthy volunteers (407 ± 589 pg/ml). Serum bFGF levels were also significantly (P < 0.05) elevated in the patients with thymic carcinoma (2 740 ± 631 pg/ml) compared with those in the healthy volunteers (1 728 ± 1 192 pg/ml). However, the serum VEGF and bFGF levels did not significantly differ between the patients with thymoma and the healthy volunteers. Serum VEGF and bFGF levels did not significantly differ according to the stage and pathological subtype of thymoma. Moreover, there was no correlation between the serum levels of VEGF and those of bFGF. Thus, while serum VEGF and bFGF levels may serve as markers for thymic epithelial tumors, it is unlikely that circulating VEGF and bFGF could be used as markers for assessing the progression of thymoma tumors. Received: November 10, 2000 / Accepted: May 15, 2001     

Clinical Significance of K-Ras Mutations in Intraoperative Tumor Drainage Blood From Patients With Colorectal Carcinoma

Background: Recurrent and metastatic carcinoma of the colorectum remains a major problem. This may be ascribed to the presence of micrometastasis at diagnosis. The purpose of this study was to analyze prospectively the clinical value of detecting K-ras mutations in the perioperative circulating blood from patients with colorectal carcinoma.Methods: Twenty-four patients whose tumor carried mutations in codon 12 of the K-ras gene were studied for the presence of cancer cells in perioperative blood samples, in particular, tumor drainage samples. A detection assay using CD45 immunomagnetic separation plus nested mutant allele specific amplification (MASA) was performed.Results: K-ras mutations in CD45 negative cells in tumor drainage blood were detected in 7 (29.2%) of 24 patients. There was no significant relationship between the presence of a K-ras mutation and clinicopathological features. Four (57.1%) of the seven patients with a positive K-ras mutation in drainage blood had early recurrent disease. Of the 17 patients with no K-ras mutation, none developed metastatic disease. The recurrence rate of the K-ras mutation positive group was higher than that of the K-ras mutation negative group (P < .01). There was a significant difference, regarding prognosis, between K-ras mutation positive and negative groups (P < .01).Conclusions: This preliminary study demonstrates that the detection of circulating cancer cells in the tumor drainage blood by our new assay system may provide a predictor of recurrence and metastasis of colorectal cancer.     

新生淋巴管与肺癌转移相关性研究进展

新生淋巴管形成对肺癌的转移起着积极的促进作用。目前，新生淋巴管形成是肿瘤转移研究中的一个热点，在肿瘤转移中所起的作用正受到越来越多的关注。肺癌淋巴结转移是肺癌患者预后不良的重要因素，新生淋巴管与肺癌转移之间的关系现还有许多不明之处。现对其研究进展进行综述。     

A prospective analysis of plasma endostatin levels in colorectal cancer patients with liver metastases

Background: Circulating inhibitors of angiogenesis have been suggested to affect the growth of distant micrometastatic disease in patients with cancer. This study was designed to evaluate circulating endostatin levels in colorectal cancer patients with liver metastases.Methods: Plasma samples from 30 colorectal cancer patients with liver metastases were analyzed for endostatin and vascular endothelial growth factor (VEGF) by using competitive enzyme immunoassays. Samples were compared with plasma from age- and sex-matched healthy controls; values >2 SD above the control mean were considered elevated.Results: Plasma endostatin levels were significantly higher in the 30 cancer patients than controls (P < .0001) and correlated with preoperative VEGF levels (P = .0008). Eighteen patients underwent surgical treatment (liver resection, n = 10; or isolated hepatic perfusion with melphalan, n = 8). Seventeen treated patients were available for follow-up. Eight of 11 patients who progressed had elevated plasma endostatin levels at the time of progression. None of six patients who remained progression free had elevated endostatin levels at last follow-up (P = .02).Conclusions: Plasma endostatin levels are elevated in colorectal cancer patients with liver metastases and correlate with VEGF levels. Elevated endostatin levels during follow-up are associated with disease progression. Understanding the role of endogenous endostatin in cancer patients may lead to novel strategies to inhibit tumor angiogenesis.     

人胃癌VEGF和NOS的表达与肿瘤血管生成的关系

目的　研究血管内皮生长因子 (VEGF)、诱导型一氧化氮合酶 (iNOS)及内皮型一氧化氮合酶 (eNOS)在人胃癌表达的相关性及与胃癌血管生成的关系 ,探讨NO和VEGF的相互作用及NO在VEGF促肿瘤生长中的作用机理。方法　应用免疫组化方法检测 34例胃癌组织中VEGF、iNOS和eNOS的表达及分布 ,用血管内皮细胞第Ⅷ相关抗原 (FⅧRAg)行免疫特异性染色计数肿瘤微血管密度 (MVD)。 结果　 34例胃癌组织中 ,表达iNOS者占 73.5 % ,表达eNOS者占 82 .4 % ,表达VEGF者占 91.2 % ;VEGF与iNOS的表达具有相关性 (P<0 .0 0 5 ) ,VEGF与eNOS的表达则无相关性 (P>0 .0 5 ) ;表达VEGF的胃癌其MVD明显高于不表达VEGF的胃癌 (P<0 .0 2 5 ) ,表达iNOS的胃癌其MVD明显高于不表达iNOS的胃癌 (P<0 .0 5 ) ,表达eNOS的胃癌MVD与不表达eNOS的胃癌差异无显著性意义 (P>0 .0 5 )。结论　胃癌组织中MVD随着VEGF和iNOS表达的增加而增加 ,提示两者对胃癌的血管生成起促进作用 ;VEGF与iNOS的表达具有相关性 ,提示iNOS在VEGF的生成和发挥作用过程中起重要作用。     

大肠癌患者VEGF和Endostatin的表达及其与临床病理特征的关系

目的：探讨大肠癌患者手术前后血清血管内皮生长因子（VEGF）和Endostatin的动态变化规律及其与临床病理特征的关系。方法：ELISA法检测大肠癌患者（大肠癌组）术前及术后2周血清VEGF和Endostatin水平,并与大肠腺瘤患者（大肠腺瘤组）和健康对照者（对照组）进行比较。结果：1）大肠癌组术前血清VEGF水平显著高于大肠腺瘤组及对照组（P均〈0.01）。2）大肠癌组术前血清Endostatin水平显著高于大肠腺瘤组及对照组（P均〈0.01）。3）大肠癌组术前血清VEGF、Endostatin水平与原发肿瘤大小、细胞分化程度、区域淋巴结转移、肝转移及Dukes分期密切相关（P均〈0.05）,与性别、肿瘤部位等因素无关（P〉0.05）。4）大肠癌组术后2周血清VEGF水平较术前显著下降,而血清Endostatin水平较术前升高（P均〈0.01）。结论：大肠癌患者血清VEGF和Endostatin水平升高,且与原发肿瘤大小、细胞分化程度、区域淋巴结转移、肝脏转移及Dukes分期等因素有密切关系;血清VEGF和Endostatin水平是评价大肠癌恶性行为、预测浸润和转移程度的有效指标。     

Markedly Elevated Levels of Vascular Endothelial Growth Factor in Malignant Ascites

Background: Vascular endothelial growth factor (VEGF) is a potent angiogenic factor that also has the ability to increase vascular permeability. Malignant ascites has significant morbidity, but the mechanism of its development is unknown. Because of the permeability-inducing properties of VEGF, we hypothesized that malignant ascites formation is associated with high levels of VEGF. The purpose of our study was to determine the role of VEGF in malignant ascites formation.Methods: Ascites from 25 patients with gastric (n = 6), colon (n = 7), or ovarian (n = 12) cancers was collected by paracentesis or surgery. VEGF protein levels were determined by enzyme-linked immunosorbent assay. The effect of ascites on endothelial cell permeability was assessed by evaluating propidium iodide uptake by human umbilical vein endothelial cells (HUVECs) exposed to ascites. Neutralizing antibodies to VEGF added to ascites were used to determine the causal effect of VEGF in permeability induction.Results: VEGF protein levels were markedly increased in malignant ascites compared with levels in nonmalignant cirrhotic ascites (controls). VEGF protein levels in ovarian, gastric, and colon cancer ascites were found to be increased 45, 23, and 12 times, respectively, compared with levels in cirrhotic ascites. Malignant ascites from patients with colon and gastric cancer caused an increase in permeability in HUVECs in all cases. Neutralizing VEGF activity in colon cancer ascites decreased in-vitro HUVEC permeability in three of four cases.Conclusions: VEGF protein levels are markedly elevated in malignant ascites. VEGF may play a role in malignant ascites formation by increasing endothelial cell permeability.     

大肠癌细胞和外周血淋巴细胞对化疗药物的体外敏感性

目的:探讨大肠癌细胞及外周血淋巴细胞对化疗药物体外敏感性的及二者的相关性。方法:用MTT法检测40份大肠癌标本及外周血淋巴细胞对氟尿嘧啶(5-FU)、奥沙利铂(L-OHP)、伊立替康(CPT)单药、两药及三药(全量或半量)应用的敏感性。结果:单药最有效的药物依次为伊立替康、奥沙利铂和氟尿嘧啶,敏感率分别为35.0%、27.5%和20.0%;三药联合应用的抑制效果显著优于两药联合(P<0.05),三药全量及半量联合应用效果差异无显著性(P>0.05);癌细胞及外周血淋巴细胞对3种化疗药物的敏感性有很好的相关性(r=0.969)。结论:MTT可用于为大肠癌患者选择合适的化疗药物,由氟尿嘧啶、奥沙利铂及伊立替康的联合应用具有高效协同抑制大肠癌细胞的作用。     

结直肠腺瘤及腺癌中P73表达与VEGF的关系

目的:探讨结直肠腺瘤和腺癌组织中P73的表达及其与血管内皮生长因子(vascular endothelial growth factor,VEGF)的关系.方法:采用免疫组化(SP法),检测P73抗体与VEGF蛋白的表达.结果:结直肠腺瘤中18例P73表达有17例(94.4%)阳性反应,与结直肠腺癌组间无显著性差异(P＞0.05),而与癌旁正常组织间存显著性差异(P＜0.01);同一标本中,癌组织中P73的染色强度均大于其癌旁组织(P＜0.01).腺癌各组VEGF阳性表达率高于腺瘤(P＜0.01), 整个腺癌组VEGF阳性表达率显著高于腺瘤组(84.1%∶33.3%,χ2=3.911,P＜0.05).结论:结直肠腺瘤及腺癌中P73与VEGF有显著的相关性, 加强术前检测P73,对高表达的患者有助于结直肠腺瘤癌变或腺癌早期诊断,及指导术后治疗.     

胃癌组织NOS和VEGF表达与癌细胞增殖相关性的研究

目的:研究人胃癌组织血管内皮生长因子(VEGF)、诱导型一氧化氮合酶(iNOS)、内皮型一氧化氮合酶(eNOS)的表达与癌细胞增殖的相关性。方法:应用免疫组化方法检测34例胃癌手术切除标本VEGF、iNOS、eNOS和增殖细胞核抗原(PCNA)的分布及表达。结果:①31例胃癌组织表达VEGF,25例(73.5%)表达iNOS,28例(82.4%)表达eNOS;②VEGF与iNOS的表达具有明显相关性,VEGF与eNOS的表达无明显相关性;③表达VEGF的胃癌PCNA标记指数(PLI)明显高于不表达VEGF的胃癌,表达iNOS的胃癌PLI明显高于不表达iNOS的胃癌,表达eNOS的胃癌PLI与不表达eNOS的胃癌无显著性差异(P<0.05)。结论:①VEGF与iNOS的表达具有明显相关性,说明iNOS在VEGF的生成和发挥作用过程中起重要作用;②PLI随着VEGF和iNOS表达的增加而增加,说明二者对胃癌细胞增殖具有促进作用。     

Vascular Endothelial Growth Factor C Promotes Lymph Node Metastasis in a Rectal Cancer Orthotopic Model

Purpose Vascular endothelial growth factor C (VEGF-C), a novel member of the vascular endothelial growth factor family, is a relatively specific lymphangiogenic growth factor. It has been suggested that increased expression of VEGF-C in primary tumors is correlated with lymph node metastasis. We conducted this study to determine whether VEGF-C directly affects lymphangiogenesis and lymph node metastasis in colorectal cancer.Methods For an accurate analysis and clear visualization of metastases, the rectal cancer cell line, DLD1, was engineered to stably express green fluorescent protein (GFP) (DLD1/GFP). We implanted DLD1/GFP cells overexpressing VEGF-C orthotopically into the rectal walls of nude mice.Results Lymph node metastasis was confirmed in all (100%) of the mice bearing DLD1/GFP-VEGF-C tumors, but in only 25% of the mice bearing control tumors. There were more lymph node metastases per mouse in the mice bearing DLD1/GFP-VEGF-C tumors than in the mice bearing control tumors. There were no differences in cell growth and motility in vitro or in the resulting tumor volume from the implanted cells between the two groups. Immunohistochemical staining revealed that VEGF-C induced the growth of lymphatic vessels, which were enlarged in the tumor periphery and contained tumor cell emboli.Conclusion These results suggest that VEGF-C-induced lymphangiogenesis mediates tumor spread and the formation of lymph node metastasis.     

Expression of CD44, Vascular Endothelial Growth Factor, and Proliferating Cell Nuclear Antigen in Severe Venous Invasional Colorectal Cancer and Its Relationship to Liver Metastasis

(Received for publication on Jan. 5, 1999; accepted on Nov. 11, 1999)     

Expression of Vascular Endothelial Growth Factor (VEGF) and Epidermal Growth Factor Receptor (EGFR) is an Independent Prognostic Indicator of Worse Outcome in Gastric Cancer Patients

BACKGROUND: Unlike other human tumors, gastric cancer remains a great therapeutic challenge since no standardized postoperative treatment exists. Knowledge of molecular pathways determining the behavior of individual gastric tumors seems to be crucial for therapeutic decisions, and evaluation of vascular endothelial growth factor (VEGF) and epidermal growth factor receptor (EGFR) expression might be critical for prognosis, assessment, and identification of patients that could be treated with tailored therapies. METHODS: VEGF and EGFR determination was performed in 88 gastric cancer samples as well as 25 normal gastric mucosa specimens from non-cancer patients using a commercially available immunohistochemistry kit. In all samples, the correlation of VEGF and EGFR expression was investigated with each other, and with other prognostic indicators in all samples, and, finally, with survival rates in 69 patients undergoing potentially curative surgery. RESULTS: Forty-eight per cent (42 cases) of gastric cancers expressed VEGF, and 44% (39 cases) stained for EGFR. In curatively treated patients, VEGF and EGFR expression was demonstrated to correlate with worse survival in both univariate and multivariate analyses. Molecular profiling was shown to more accurately estimate the risk of cancer-related death than TNM stage, and, of most interest, to allow sorting out high-risk patients within the same stage. CONCLUSIONS: These findings provide evidence that contemporary evaluation of VEGF and EGFR expression may be crucial to select gastric cancer patients with poor prognosis who may benefit of tailored treatments.     

HGF/SF对姜黄素诱导大肠癌细胞凋亡的作用

目的 观察肝细胞生长因子／离散因子（HGF／SF）在姜黄素诱导的大肠癌细胞凋亡中的作用。方法 采用MTT法分析姜黄素对大肠癌细胞的抑制作用；流式细胞术评价HGF抗凋亡作用。结果不同浓度姜黄素作用Caco2细胞后发现，只在64μg／ml浓度发挥作用，即抑制细胞增殖；而同时加入不同浓度的HGF后可以拮抗姜黄素引起的Caco-2细胞生长的抑制作用，但HGF发挥作用无浓度依赖关系。流式细胞术检测结果发现，姜黄素只在64μg／ml浓度下诱导Caco-2细胞凋亡，在加入不同浓度HGF后，凋亡明显减少，但并无剂量依赖关系。MAPK途径在HGF拮抗姜黄素诱导的Caco-2细胞凋亡中的作用发现，阻断p42／p44MAPK和p38MAPK后，并不影响HGF对Caco-2细胞凋亡的保护作用。结论 HGF拈抗姜黄素诱导的Caco-2细胞凋亡，MAPK信号传导途径可能不参与此过程。