Comparison of Fluorescence in situ Hybridization and Immunohistochemistry for Assessment of HER-2 Status in Breast Cancer Patients

The accurate assessment ofa proto-oncogene, human epidermal growth factor receptor-2 gene （HER-2）, is extremely important for the therapy and prognosis of breast cancer. Currently, immunohistochemistry （IHC） is the method widely used for the detection of HER-2 protein. Fluorescence in situ hybridization （FISH） has been suggested to be a golden standard assay for HER-2 amplification. This study examined the expression and amplification of HER-2 in paraffin-embedded sections of breast cancer tissues, and compared the two methods on the measurement of HER-2 status. HER-2 gene and protein were determined in breast cancer samples from 52 Chinese women by FISH and IHC respectively. The findings indicated that the HER-2 gene amplification was found in 18 cases （34.6%） by FISH and the HER-2 protein over-expression （score 3＋） in 15 cases （28.8%） by IHC. hnmunohistochemically, 28.6% of the cases scored as 2＋ and 93.3% of the cases scored as 3＋ were HER-2-positive by FISH. There was a significant correlation between the HER-2 gene amplification and HER-2 protein over-expression in breast cancer （P〈0.005）. No correlation was noted between the HER-2 gene amplification and any of the clinicopathological parameters examined, including age, menopausal status, menarche age, tumor size, histological tumor type, histological grade, lymph node status, and the expression of ER and PR. It was concluded that the detection of HER-2 gene amplification in breast cancer by FISH is valuable and can compare with HER-2 protein detection by IHC.

DISTRIBUTION OF VEGF mRNA IN BREAST CANCER WITH NONRADIOACTIVE IN SITU HYBRIDIZATION AT ELECTRON MICROSCOPIC LEVELS

Object: To localize the mRNA coding for VEGF at Ultrastractural level in human breast cancer by using digoxigenin-labeled cDNA probes. Methods: Nonradioactive in situ hybridization at electron microscopic level was employed to detected VEGF mRNA in breast cancer. Result: Cancer cells and endothelial cell of angiogensis show dark color in experiment sections. No dark color can be found in control sections. Positive hybridization signals showed dark dot and were located in various compartments of the breast cancer cell and endothelial cell in experiment section. No labeling was observed in control sections. In experiment sections, the staining appeared concentrated in cytoplasm and nucleus of the breast cancer cell and endothelial cell. Conclusion: Nonradioactive in situ hybridization at electron microscopic level is efficient for direct observation of the target site mRNA of VEGF in the cytoplasm and nucleus.

Decorin mRNA和p57^(KIP2) mRNA在非小细胞肺癌中的表达及其临床意义

目的检测Decorin mRNA和p57^(KIP2) mRNA在非小细胞肺癌中的表达情况,并分析它们与非小细胞肺癌的病理、临床特征及预后的关系。方法选取石蜡包埋的59例非小细胞肺癌及12例癌旁正常肺组织的标本制作组织芯片。利用原位杂交法检测Decorin mRNA和p57^(KIP2) mRNA的表达情况。结果Decorin mRNA在癌旁正常肺组织和非小细胞肺癌组织中的表达率分别为66.7%(8/12)和20.3%(12/59),正常组织中Decorin mRNA的表达高于肺癌组织(P<0.05)。大细胞肺癌中Decorin mRNA的表达高于鳞癌(P<0.05)。鳞癌中p57^(KIP2) mRNA的阳性表达率为63.2%(12/19),高于腺癌、大细胞癌和小细胞癌(P<0.05)。在无淋巴结转移的非小细胞肺癌标本中Decorin mRNA和p57^(KIP2) mRNA的表达呈增高趋势,与有淋巴结转移的标本比较其差异具有统计学意义。Decorin mRNA和p57^(KIP2) mRNA的表达无相关性(P>0.05)。结论Decorin和p57^(KIP2)的表达可能与非小细胞肺癌的组织学类型和转移有关,有望成为监测非小细胞肺癌患者病情发展及评价预后的有用指标。

Comparison of immunohistochemistry with fluorescence in situ hybridization in determining the human epidermal growth factor receptor 2 status of breast cancer specimens： a multicenter study of 3149 Chinese patients

Background Accurate detection of human epidermal growth factor receptor 2 （HER2） expression and gene amplification is crucial for the application of HER2-specific therapy and for evaluating the response of patients with breast cancer.A uniform and standard procedure of immunohistochemistry （IHC） and fluorescence in situ hybridization （FISH） needs to be established for evaluating the HER2 status in breast cancer tissues for the treatment of patients with real HER2-positive tumors.The present multicenter study was aimed to examine the HER2 status in breast cancer specimens from Chinese patients using both IHC and FISH methods.Methods A multicenter study was performed on the HER2 status in 3 149 breast cancer specimens from different ethnic populations and areas in China by IHC and FISH assays.The potential association of HER2 status with demographic and clinical characteristics was analyzed.Results The positive rates for HER2 over-expression and HER2 amplification were 23.3％ and 27.5％ in this study,respectively.The concordance between IHC and FISH was 71.2％ （K=0.494,P ＜0.001）.Furthermore,72.9％ of specimens with IHC 2＋ were negative to FISH.The discordance rates among laboratories were from 5％ to 28％ for IHC and 1％ to 16％ for FISH.HER2 amplification was associated significantly with advanced tumor stage （Ⅲ or Ⅳ,P=0.002）,large tumor size （＞5 cm,P=0.002）,moderate and poor histological grades （P ＜0.0001）,post-menopause （P ＜0.0001）,ER-PR-（P=0.002）,and having ＞4 lymph nodes affected （P ＜0.0001） in this population.The positive rates of HER2 amplification in specimens from Man and Hui Chinese were significantly higher than that in other Chinese populations.There are slightly higher positive rates of HER2 expression and amplification in Chinese patients with breast cancer.Conclusion These findings may provide new insights into understanding the epidemiological features of HER2 expression and amplification,and may be valuable for clinical practice.

Correlations of β-catenin,Ki67 and Her-2/neu with gastric cancer

Objective:To study the clinical pathologic characteristics ofβ-catenin,Ki67 and Her-2/neu in gastric cancer and the correlation ofβ-catenin and Ki67 to the protein expression and gene conditions of Her-2/Neu.Methods:The protein expression ofβ-catenin,Ki67 and Her-2/Neu was detected by immunohistochemistry in 101 cases of gastric cancer and the gene conditions of Her-2/Neu by fluorescence in situ hybridization(FISH).Results:The protein expression ofβ-catenin,Ki67 and Her-2/Neu had close relationship with the clinical pathologic characteristics of gastric cancer.Theβ-catenin and Ki67 had obvious correlation to the differentiation,infiltration and lymphatic metastasis of the gastric cancer(P<0.05).The Ki67 had close relationship with the tumor-node-metastasis staging staging of gastric cancer(P<0.05).Her-2/Neu had close relationship with the differentiation and tumor-node-metastasis staging of gastric cancer(P<0.05)but had no relationship with the infiltration and lymphatic metastasis of the gastric cancer(P<0.05).The protein expression of Ki67 had significantly positive correlation to the protein expression and gene amplification conditions of Her-2/Neu(r=0.567,P<0.05 for protein;r=0.304,P<0.05 for gene).Conclusions:Combined detection ofβ-catenin,Ki67 and Her-2/Neu can be used as a reliable method to help the observation of biological behavior,diagnosis and prognosis of gastric cancer,and Ki67 can be used to serve the preliminary screening of Her-2/Neu gene state.

Polypeptide Daintain as a New Biomarker for Detecting Breast Tumor

Daintain, a novel bioactive peptide produced and secreted by macrophages, was expressed in breast tumor tissues. The spatial distributions of daintain in 66 breast tumor specimens were investigated with immuno-histochemistry method. Reverse transcription polymerase chain reaction （RT-PCR） and in situ hybridization inspection system were also used to detect daintain in 45 cases of malignant breast tumors. The final results show that 93% high positive responses to daintain on breast cancer tumors. RT-PCR demonstrated that, no smear of daintain transcripted in benign tissues was found, and light smear in peri-cancer tissue was observed. Distribution of daintain was distinguishable among benign tissues, hyperplasia tissues, immature hyperplasia and invasive breast cancer, which can be used to mark the progression of the malignant lesion development. We conclude that the expression of daintain is up-regulated in breast cancers, which indicates that the peptide is closely associated with the disease progression. So daintain could be used as the biomarker for detecting breast cancer.

HER-2 expression after neoadjuvant chemotherapy of the breast cancers

Objective:The aim of this study was to study changes of HER-2 expression after neoadjuvant chemotherapy in the breast cancer cases.Methods:One hundred and thirty-seven female patients with primary breast cancers,who received neoadjuvant chemotherapy,underwent core needle puncture and Mammotome biopsy before chemotherapy,and the biopsy results were used as the basis of histological diagnosis,fluorescence in situ hybridization (FISH) was performed to test HER2 status of tumor tissues before and after chemotherapy.All patients underwent FEC,TE,or AC neoadjuvant chemotherapy of 2-6 cycles before surgery.Results:Twenty-two patients were positive according to FISH test among 137 preoperative patients,8 patients achieved pathological complete remission after chemotherapy (three HER-2 positive patients and five negative patients),91 patients achieved partial remission,24 patients were stable,and 14 cases were invalid.Twenty-two patients were positive according to FISH test (8 patients with pathological complete remission did not undergo test),and positive patients still expressed positively after chemotherapy before neoadjuvant chemotherapy.Three negative patients were converted to be positive,and changes before and after chemotherapy had no statistical difference (P>0.05).Conclusion:Neoadjuvant chemotherapy makes no influence on patients with HER-2 positive expression,while patients with negative expression can be converted to be positive,but without significant difference.

Predictive biomarker and clinicopathological characteristics analysis for recurrence of early gastric cancer

Objective: The aim of this study was to identify the correlation between the clinicopathological characteristics and recurrence in early gastric cancer (EGC), what's more, we attempt to look for a predictive biomarker to predict and treat for re-currence of EGC. Methods: This study retrospectively analyzed 178 early gastric cancer patients who had the complete post-operative and follow-up medical records in the First Affiliated Hospital of Yangtze University (China) between January 1995 to December 2005. All of them were followed-up to December 2009 regularly. Computer tomography (CT), endoscopy, and single photon emission computed tomography (SPET-CT) were used to diagnose for recurrence of EGC. Immunohistochem-istry (IHC) and fluorescence in situ hybridization (FISH) were used for the detection of cerbB2. Chi-square test was applied to this study for statistics analysis. Results: Fourteen patients had recurrence. Eighteen patients were cerbB2-positive, including twelve recurrence patients and six norecurrence patients. Sex, tumor depth, and lymph node metastasis were related to the recurrence of EGC. Also, cerbB2-positive patients had the higher recurrence rate compared to the cerbB2-negative patients. Conclusion: Recurrence of EGC after curative resection can be predicted by using some clinicopathological characteristics. CerbB2 can be used as a predictive biomarker for recurrence of EGC.

Immunohistochemical Biomarkers in Ductal Carcinoma <i>In Situ</i>

Introduction: Breast ductal carcinoma In Situ (DCIS) can be defined as a malignant epithelial proliferation with growth limited by the basal membrane of the ductal epithelium, with no evidence of stromal invasion. There has been a trend of trying to subcategorize DCIS based on cell proliferation assays (Ki67) and the expression of hormone receptors and the human epidermal growth receptor (HER-2) as detected by immunohistochemistry, similar to invasive breast carcinomas (IBC). The aims were to evaluate the expression of breast cancer marker proteins in DCIS by immunohistochemistry to better categorize it. Methods: 46 biopsies from women with DCIS and IBC Luminal A-like were evaluated by immunohistochemistry staining of proteins already known to be biomarkers in IBC. For controls, normal breast tissue from mammoplasty (n = 3) was used. Results: Our results showed an increase of estrogen receptor (ER) and progesterone receptor (PR) expression relative to that in normal tissue samples (p < 0.0001). No differences in steroid hormone expression patterns were seen between DCIS and IBC tumors (p = 0.3145;p = 0.7341, respectively). The proliferation levels of the DCIS and IBC samples were similar as evaluated by the Ki67 labeling index. Only 12.90% of samples showed amplification of HER-2. Conclusion: The biology of DCIS is not well understood given the complexity and heterogeneity of the disease, which makes it important to better sub-categorize this tumor, especially considering the possibility of identifying DCIS cases with the potential for recurrence and evolution into IBC.

Snail基因及其蛋白、E-cadherin蛋白在胃癌中的表达及其临床意义

目的观察Snail mRNA及其蛋白、E-cadherin蛋白在胃癌组织中的表达及其与胃癌临床病理特征的关系,并探讨它们在胃癌发生、发展中的作用及其临床应用价值。方法收集96例手术切除胃癌标本,同时取80例癌旁组织作为对照。应用免疫组织化学S-P法检测胃癌组织、癌旁组织中snail蛋白、E-cadherin蛋白的表达;运用原位分子杂交技术检测胃癌组织、癌旁组织中Snail mRNA的表达。结果(1)Snail蛋白在胃癌组织阳性率(83.3%)显著高于癌旁组织(41.25%)(P<0.05);高、中分化组Snail蛋白阳性表达率显著低于低分化组(P<0.05);Snail蛋白的阳性表达率在乳头状腺癌、管状腺癌及低分化腺癌与黏液癌之间差异有显著性(P<0.05);Snail蛋白的表达与胃癌浸润深度、淋巴结转移及远处转移有关(P<0.05),与性别、年龄、肿瘤大小、肿瘤部位及临床分期无关(P>0.05);(2)胃癌组织中snail mR-NA的阳性率(76%)显著高于癌旁组织(30%)(P<0.05);高、中分化组Snail mRNA阳性表达率显著低于低分化组(P<0.05);Snail mRNA的阳性表达率在乳头状腺癌、管状腺癌及低分化腺癌与黏液癌之间差异有显著性(P<0.05);Snail mRNA的表达与浸润深度及淋巴结转移有关(P<0.05),与性别、年龄、肿瘤大小、肿瘤部位、临床分期及远处转移无关(P>0.05);(3)E-cadherin蛋白在胃癌组织阳性率(37.5%)显著低于癌旁组织(100%)(P<0.05);高、中分化组E-cadherin蛋白阳性率显著高于低分化组(P<0.05);E-cadherin蛋白阳性率在乳头状腺癌、管状腺癌及低分化腺癌与黏液癌之间差异有显著性(P<0.05);E-cadherin蛋白的表达与胃癌浸润深度、淋巴结转移、临床分期及远处转移有关(P<0.05),与性别、年龄、肿瘤大小、肿瘤部位均无关(P>0.05);(4)胃癌组织中snail mRNA和snail蛋白的表达呈正相关(r=0.594,P<0.05);Snail蛋白和E-cadherin蛋白的表达呈负相关(r=-0.234,P<0.05)。结论(1)E-cadher-in蛋白低表达与Snail蛋白高表达可能是胃黏膜恶性转变以及胃癌发生浸润转移的重要生物学标志;联合检测E-cadherin蛋白与Snail蛋白对预测胃癌浸润转移有重要意义。(2)Snail蛋白可能在转录水平上调控E-cadherin蛋白的表达。

Tenascin-C在乳腺导管原位癌的表达及其意义

目的:探讨Tenascin-C(Tn-C)在乳腺导管原位癌中的表达及其意义。方法:应用免疫组化S-P方法检测52例乳腺导管原位癌Tn-C的表达,及其与ER、PR和HER-2等表达的相关性。结果:Tn-C表达阳性率为60%,其中Van-Nuys1组的阳性率为38%,Van-Nuys2组的阳性率为70%,Van-Nuys3组的阳性率为88%,组间阳性表达差异有统计学意义(P<0.01)。Tenascin-C的阳性表达和ER、PR阴性表达及HER-2阳性表达呈正相关(P<0.05)。结论:Tn-C可以作为判断乳腺导管原位癌恶性程度、预后的一个指标。

Stathmin在胰腺癌中的表达及意义

目的探讨Stathmin在胰腺癌中的表达水平以及与临床、病理特征方面的关系。方法通过免疫组化法和原位杂交技术分别检测40例胰腺癌癌组织、27例胰腺癌旁正常组织中Stathmin及其mRNA的表达情况。结果 Stathmin在胰腺癌癌组织的阳性率显著高于胰腺癌旁正常组织(77.5%vs 7.4%),差异有统计学意义(P<0.05);Stathmin mRNA的表达阳性率也明显高于癌旁正常组织(80.0% vs7.4%),差异有统计学意义(P<0.05)。不同分化程度、有无淋巴结转移、有无远处转移以及TNM分期等方面胰腺癌组织之间的Stathmin表达差异均有统计学意义(P<0.05)。结论 Stathmin及其mRNA在胰腺癌组织的高水平表达,与胰腺癌的分化程度、淋巴结转移、远处转移及TNM分期密切相关,提示Stathmin可能参与了胰腺癌发生、发展,Stathmin对胰腺癌的早期诊断及预后判断有重要意义。

AnnexinⅡ、S100A4在甲状腺乳头状癌中的表达及意义

目的探讨甲状腺乳头状癌中钙磷脂结合蛋白Ⅱ(AnnexinⅡ)与钙结合蛋白S100A4的表达,分析其与甲状腺乳头状癌发生及淋巴结转移的关系以及二者的相关性。方法应用免疫组织化学SP法和原位杂交技术检测50例甲状腺乳头状癌组织、14例甲状腺腺瘤组织、26例结节性甲状腺肿组织和10例正常甲状腺组织中AnnexinⅡ、S100A4蛋白和AnnexinⅡmRNA的表达。结果甲状腺乳头状癌中AnnexinⅡ、S100A4蛋白和AnnexinⅡmRNA的阳性表达率分别为82.00%(41/50)、78.00%(39/50)和74.00%(37/50),均明显高于甲状腺腺瘤组织、结节性甲状腺肿组织及正常甲状腺组织(P<0.05);三者在癌组织中的表达与甲状腺乳头状癌淋巴结转移有关(P<0.05);AnnexinⅡ蛋白与S100A4蛋白的表达呈正相关性(r=0.380,P=0.025);AnnexinⅡ蛋白与AnnexinⅡmRNA的表达呈正相关性(r=0.434,P=0.008)。结论 AnnexinⅡ蛋白、AnnexinⅡmRNA及S100A4蛋白的高表达与甲状腺乳头状癌的发生和淋巴结转移密切相关,二者联合检测有助于鉴别甲状腺良恶性病变,并预测肿瘤的发展和预后。

E-cadherin与乳腺癌分化程度及其淋巴结转移的关系研究

目的 探讨E -cadherin基因表达在乳腺癌侵袭和转移中的作用。方法 采用原位杂交技术检测 30例正常乳腺组织和 4 8例乳腺癌原发肿瘤及其淋巴结转移灶中E -cad herinmRNA表达情况。结果 乳腺癌原发肿瘤组织和淋巴结转移灶中 ,E -cadherinmRNA阴性表达率 (分别为31.3% ,4 6 .7% )与正常乳腺组织之间的差异非常显著 (P <0 .0 0 1)。E -cadherinmRNA表达强度在乳腺癌Ⅰ～Ⅲ级之间及有无淋巴结转移的原发肿瘤之间具有显著性差异 (P <0 .0 5 )。结论 肿瘤抑制基因E

Survivin在人乳腺浸润性导管癌中的表达及意义

目的 :探讨Survivin在人乳腺浸润性导管癌中的表达及临床意义。方法 :用原位杂交、免疫组化对人乳腺浸润性导管癌未化疗组 2 8例、化疗组 17例、癌旁正常组织 15例、乳腺良性病变 2 0例检测。结果 :每指标的SurvivinmRNA与Survivin蛋白表达无显著性差异且正相关 ;Survivin在人乳腺浸润性导管癌未化疗组表达高 ,不同年龄、TNM分期、病理分级、淋巴结转移、受体类型组间有显著性差异 (P <0 .0 1) ;化疗组比未化疗组低 (P <0 .0 1) ;在癌旁正常组织中无表达 ,在乳腺良性病变中低表达。结论 :Survivin在人乳腺浸润性导管癌高表达 ,有利于其诊断及疗效的判定。

β-catenin在大肠癌的异常表达及其意义

目的研究β-catenin在大肠癌的异常表达及其意义。方法应用原位杂交方法检测β-catenin mRNA和免疫组化SP法检测β-catenin蛋白在正常黏膜、非瘤性息肉、腺瘤和大肠腺癌等不同大肠病变组织中的表达情况,分析其之间的相互关系。结果β-catenin mRNA和蛋白在大肠癌的发生过程中均有高表达,β-catenin蛋白在癌变过程中正常的细胞膜表达逐渐缺失,同时表现为异位高表达。结论由于β-catenin mRNA和蛋白的高表达以及β-catenin蛋白在细胞浆积聚并逐步进入细胞核最终导致细胞癌变。

survivin和hTERT在大肠黏膜癌变中表达及其与c-myc关系

目的 研究survivin和端粒酶催化亚单位 (hTERT)在大肠黏膜癌变中表达及其与c myc关系。方法 采用原位杂交和免疫组化 (SP法 )检测hTERTmRNA、survivin、c myc蛋白在 45例大肠腺癌、2 0例腺瘤、10例正常组织的表达 ,并对各指标阳性率和相关性分析。结果 肠腺癌中survivin阳性率 66 7% ,hTERTmRNA85 % ,c myc 5 3 % ,三者在腺瘤中分别为 3 0 %、45 %、2 0 % ,正常黏膜为0 %、10 %、0 % ,survivin和hTERTmRNA表达与大肠癌的分化程度及临床参数无明显相关性 ,与c myc明显相关 ,survivin与hTERTmR NA呈正相关性 ( χ2 =10 .2 3 5 ,P =0 0 0 1)。结论 survivin和hTERT在大肠癌中高表达 ,非癌组织中低表达 ,可成为诊断治疗的新靶点 ,但不能作为肿瘤进展的标志物 ,survivin、hTERT、c

FGFR-1、cyclinD1在乳腺癌中的表达及其临床意义

目的探讨成纤维生长因子受体(fibroblast growth factor receptor,FGFR-1)及细胞周期蛋白D1(cyclinD1)在乳腺癌组织中的表达及其临床意义。方法应用原位杂交及免疫组织化学S-P法检测乳腺癌、乳腺纤维腺瘤和正常乳腺组织中FGFR-1、cyclinD1 mRNA及蛋白水平的表达。结果在乳腺癌中,FGFR-1蛋白和FGFR-1基因的阳性率分别为70.0%(21/30)、70.0%(21/30),cyclinD1蛋白和cyclinD1基因的阳性率分别为66.7%(20/30)、76.7%(23/30),明显高于纤维腺瘤与正常乳腺组织(P<0.05),FGFR-1表达与cyclinD1表达一致。结论 FGFR-1的过表达在人类乳腺癌的发生、发展中起重要作用,其机制可能通过上调cyclinD1的表达发挥作用,FGFR-1、cyclinD1的过度表达可作为判断乳腺癌预后的重要指标。

Relationships between mucinous gastric carcinoma, MUC2 expression and survival

AIM:To investigate the expression of the four secreted gel-forming mucins(MUC2,MUCSAC,MUCSB and MUC6)in a series of gastric carcinomas,classified according Laurén's,Mulligan's,WHO and Goseki's classifications,with special attention to all the different components(major and minor)present in tumors and to follow up clinical data.METHODS:Expression of MUC2,MUC5AC,MUC5B and MUC6 was investigated using immunohistochemistry and in situ hybridization.RESULTS:Expression of secreted gel-forming mucins in gastric carcinoma was particularly complex,each mucin being not restricted to any histopathological type even considering all components(major and minor)present in a given tumor.There was a worst survival in patients with a higher content of mucus(GosekiⅡorⅣ)and high positive MUC2 expression.CONCLUSION:Complexity of mucin gene expression patterns in gastric cancer may reflect a precise state of differentiation at the cell level not recognized in used morphologic classification systems.High expression of MUC2 was nevertheless associated with mucinous subtype of the WHO classification and with groupⅡof Goseki's classification identified by the major component of a particular tumor.The quantity and quality of mucus were related to survival.

Expression of Vascular Endothelial Growth Factor C and Its Clinical Significance in Human Esophageal Squamous Cell Carcinoma

OBJECTIVE To examine the expression of vascular endothelial growth factor C （VEGF-C） in human esophageal squamous cell carcinoma （ESCC）, and to clarify its role in lymphatic metastasis in ESCC patients.METHODS Esophageal carcinoma EC9706 cells and samples from 49 patients with primary ESCC were investigated by using S-P immunohistochemistry （IHC）, the semi-quantitative reverse transcriptase-polymerase chain reaction （RT-PCR） and in situ hybridization （ISH） methods for VEGF-C expression. RESULTS VEGF-C positive expression was found in EC9706 cells through IHC, ISH and RT-PCR. Positive IHC for VEGF-C was observed in 36 of 49 cases of ESCC. There was a significant difference between the expression of VEGF-C in a lymph-node-positive group compared to a node-negative group （χ^2=4.7, P〈0.05）. Positive ISH for VEGF-C mRNA was observed in 23 of 49 cases of ESCC. There was a significant difference between the expression of VEGF-C in the lymph-node-positive group and node-negative group （χ^2=31.3, P〈0.01）. The expression of VEGF-C was significantly higher in the lymph-node-positive group compared to the node-negative group. Of 49 ESCC tissues, RT-PCR for VEGF-C mRNA was observed positively in 29 cases. There was a significant difference between the expression of VEGF-C in the lymph-node-positive group and node-negative group （χ^2=23.3, P〈0.01）. The expression of VEGF-C was significantly higher in the lymphnode-positive group compared to the node-negative group. Expressions of VEGF-C were not significantly associated with age, gender, and pathological grade. There was a relationship between VEGF-C mRNA expressions by RT-PCR and ISH （χ^2=18.5, P〈0.01） in ESCC cases, but with no significant difference between the two methods. CONCLUSION VEGF-C expression may induce lymphangiogenesis in human ESCC. There was a close correlation between VEGF-C expression and lymph node metastasis. VEGF-C can serve as a useful prognostic factor for ESCC patients.