Lymph node micrometastasis and its correlation with MMP-2 expression in gastric carcinoma

AIM： To examine matrix metalloproteinase-2 （MMP-2） expression in gastric cancer tissues and to evaluate its relationship with lymph node micrometastasis. MATERIALS： The authors studied 850 lymph nodes resected from 30 patients with gastric carcinoma who underwent gastrectomy with lymphadenetomy using reverse transcription polymerase chain reaction （RT-PCR） assay in addition to H-E staining. MMP-2 expression of the tumor tissues was detected by immunohistochemical technique （EliVision^TM plus）. RESULTS： MMP-2 expression was positive in 21 （70%） cases and negative in g （30%） cases. No significant correlations were found between MMP-2 expression and other variables such as age, gender, tumor location, tumor diameter, Lauren classification and lymphatic invasion. In contrast, MMP-2 expression correlated significantly with depth of tumor infiltration （P = 0.022）, lymph node metastasis （P = 0.030） and tumor differentiation （P = 0.043）. Lymph node micrometastases were detected in 77 （12.5%） lymph nodes of 14 （46.7%） gastric carcinoma patients. MMP-2 expression was positive in 12 （85.7%） of the 14 patients with lymph node micrometastasis, and in g （56.3%） of the 16 patients without lymph node micrometastasis （P = 0.118）. CONCLUSIONS： Our results demonstrate that MMP-2 expression has significant correlation with tumor invasion, tumor differentiation and lymph node metastases. MMP-2 expression may be an important biological characteristics and significant prognostic parameter of gastric carcinoma. We also conclude that MMP-2 may participate in the development of lymph node micrometastasis of gastric carcinoma. Further investigations are needed to draw a conclusion.

Expression of E-cadherin in gastric carcinoma and its correlation with lymph node micrometastasis

AIM: To examine the expression of E-cadherin in the primary tumor and to evaluate its relationship with lymph node micrometastasis (LNM).METHODS: The authors studied 850 lymph nodes resected from 30 patients with gastric carcinoma who underwent gastrectomy with lymphadenectomy using reverse transcription polymerase chain reaction (RT-PCR) assay in addition to H&E staining. Cytokeratin-20 (CK-20)gene marker was used in this assay. The level of E-cadherin expression in the primary tumor was examined by immunochemical technique (EliVisionTM plus).RESULTS: LNM was detected in 77 (12.5%) lymph nodes of 14 patients (46.7%) with gastric carcinoma. The incidence of LNM was significantly higher in the diffuse type (12 of 19 cases, 63.2%) than in the intestinal type of gastric carcinoma (2 of 11 cases, 18.2%, P = 0.026). The incidence of LNM also increased in accordance with the depth of tumor invasion. The loss of expression of E cadherin in primary tumors was found in 14 (46.7) of 30 tumors. The absence of E-cadherin expression was significantly associated with the Lauren classification (P = 0.026), lymph node metastasis (P = 0.011), the grade of differentiation (P = 0.004) and the lymphatic invasion (P = 0.001). Expression of E-cadherin was negative in 10 (71.4%) of the 14 patients with LNM, and in 4 (25%) of the 16 patients without LNM (P = 0.026). CONCLUSION: The current results indicate that the RT PCR assay is useful for the detection of LNM and can significantly increase the detection rate of lymph node metastasis in patients with gastric carcinoma. The Laurenclassification and depth of tumor invasion are significantlyassociated with lymph node micrometastases. Our findings also indicate that E-cadherin may play an important role in determining the growth type and differentiation of gastric carcinoma. The loss of E-cadherin expression may contribute to LNM.

Impact of lymph node micrometastasis in hilar bile duct carcinoma patients

AIM- To immunohistochemicaUy examine micrometastasis and VEGF-C expression in hilar bile duct carcinoma （HBDC） and to evaluate the clinical significance of the results. METHODS： A total of 361 regional lymph nodes from 25 patients with node-negative HBDC were immunostained with an antibody against cytokeratins 8 and 18 （CAM 5.2）, and immunohistochemical staining of VEGF-C was performed in 34 primary resected tumors. RESULTS： Lymph node micrometastasis was detected in 6 （24%） of the 25 patients and 10 （2.8%） of the 361 lymph nodes. Patients with micrometastasis showed significantly poorer survival rates than those without （P= 0.025）. VEGF-C expression was positive in 17 （50%） of 34 HBDC, and significantly correlated with lymph node metastasis （P=0.042） and microscopic venous invasion （P=0.035）. CONCLUSIONS： It is suggested that immunohistochemically detected lymph node micrometastasis has an impact on the outcome of HBDC. VEGF-C expression is highly correlated with lymph node metastasis in HBDC and might therefore be a useful predictor.

Mapping of Sentinel Lymph Node Micrometastasis in Dukes-B Colorectal Carcinoma and Its Clinical Significance

OBJECTIVE To investigate the detection of sentinel lymphnode (SLN) micrometastasis (MM) and the clinical significancein patients with Dukes-B colorectal carcinoma (CRC) using H&Estained slides.METHODS In a prospective study of 64 patients with Dukes-BCRC undergoing radical surgery,routine H&E staining combinedwith the immunohistochemical SP method was used to detect theexpression of the cytokeratin 20 (CK20) and telomerase (TLMA)in 122 SLNs from 61 of the 64 cases.During a 3-year follow up,the patients'clinicopathologic parameters data,survival and theirrelationship were collected and analyzed.RESULTS i) In 6 out of the 61 cases with successful mapping,9SLNs were positive after routine detection using H&E staining.ii) In the other 55 with negative results from H&E staining,thepositive rate of CK20 expression was 27.3% (15/55),and thepositive rate of TLMA 21.8% (12/55),after immunohistochemicalexamination (IHC) on the SLN.iii) Detection by combining the 2methods showed that the micrometastasis rate was 38.2% (21/55).iv) The recurrence or metastasis rate of CRC was significantlyhigher in the Dukes-B patients with sentinel lymph nodemicrometastasis (SLNMM) (+) than in those with SLNMM (-)(P<0.05),and simultaneously,the survival rate decreased by alarge margin in the patients with SLNMM (+),compared to thosewith SLNMM (-) (P<0.05).There were significant differences inthe recurrence or metastasis rate of cancer and the survival ratebetween the Dukes-B patients with SLNMM (-) and the Dukes-Cpatients (P<0.05).CONCLUSION Examination of anti-CK20 and anti-TLMA usingimmunohistochemical staining can be used to detect SLNMM,andthe combined examination of the 2 can increase the detection rate.The detection of SLNMM can precisely determine the Dukes stageof colorectal carcinoma,which is of help in directing postoperativeadjunctive therapy and in assessing prognosis.

Signif icance of Sentinel Lymph Node Biopsy and Immunohistochemistry in Diagnosis and Staging of Stage-cN0 Oral Squamous Cell Carcinoma

OBJECTIVE To assess the significance of sentinel lymph node biopsy （SLNB）, serial section and cytokeratin immunohistochemical staining in the diagnosis and staging of Stage-cNO oral squamous cell carcinoma （OSCC）, METHODS A blue stain, 99mTc-dextran SPECT lymphoscintigrapgy and intraoperative y-ray probes were used to examine the sentinel nodes in 31 cases with Stage-oNO oral cancer, The H＆E staining and a cytokeratin AE1/ AE3 immunohistochemistry （IHC） assessment, with serial sections, were conducted to provide results obtained from a routine pathological examination of lymph nodes, The value of the routine pathological examination of the sentinel lymph node （SLN）, serial sections and IHC determination for cervical lymph node metastasis of Stage-cN0 OSCC was appraised, RESULTS A total of 45, 55 and 51 SLNs were examined in 25 （80%）, 31 （100%） and 30 （96,5%） of the cases, by using the blue stain, y-ray probes, and SPECT lymphoscintigraphy, respectively, The average SLNs found in each case of the groups was 1,4 （1 to 3） and there were 1,302 non-NSLNs, Six positive SLN metastases were detected by routine pathological examination, among which 1 case was found to be an accompanied positive metastasis of non-SLN, One positive SLN metastasis was found after examination of serial sections plus routine H＆E staining and 2 were detected using serial sections plus AE3 immunohistochemical staining methods, No positive NSLNs were found in the study, CONCLUSION In order to make more progress in accurate SLNB diagnosis, serial sections and IHC （AE1/AE3） methods can be used for examination of the micrometastases which are difficult to identify by routine pathological sections and H＆E staining.

结肠癌前哨淋巴结微转移与FGFR4蛋白表达的关系

目的：探讨结肠癌淋巴结转移与成纤维细胞生长因子受体-4（FGFR4）蛋白的关系。方法：68例结肠癌患者，男43例，女25例，年龄34～86岁。术中染料法定位前哨淋巴结（SLN），抗CEA免疫组化法检测SLN微转移，Western Blot法测定组织内FGFR4蛋白表达。结果：每例患者的SLN数量1～4枚，平均为2.4枚（158/65）。SLN转移率为69.2％（45/65）。癌组织内FGFR4蛋白表达率为47.1％（32/68），有SLN转移者癌组织内FGFR4蛋白表达率为55.3％（26/47），无SLN转移者达率为28.6％（6/21），有显著性差别（P〈0.05）。结论：部分结肠癌组织内有FGFR4蛋白表达是结肠癌易发生淋巴结微转移原因之一。

Dukes B期大肠癌前哨淋巴结微转移的定位检测及临床意义

目的:探讨大肠癌前哨淋巴结(sentinellymph node,SLN)微转移(micrometastasis,MM)的检测方法及其临床意义。方法:前瞻性研究64例行根治性手术的Dukes B期大肠癌患者,对其中61例定位成功的122枚SLN应用常规HE染色联合免疫组化SP法,检测其前哨淋巴结中细胞角蛋白20(CK20)及端粒酶的表达,随访3年,记录患者的临床病理参数和生存资料,分析其相关性。结果:1)61例中有6例患者9枚SLN经常规HE检测阳性。2)免疫组化法检测SLN:CK20阳性表达27.3%(15/55);端粒酶阳性表达21.8%(12/55)。3)两者联合检测前哨淋巴结微转移(SLNMM)检出率为38.2%(21/55)。4)Dukes B期患者SLNMM(+)组癌复发转移率明显高于同期SLNMM(-)组(P<0.05),生存率较低(P<0.05),而与Dukes C期对比,差异无统计学意义;SLNMM(-)组患者的复发转移率、生存率与同期Dukes C期患者对比,有显著性差异(P<0.05)。结论:CK20、端粒酶免疫组化法均可检测出大肠癌SLN中存在的微转移(MM),两者联合可提高检出率;大肠癌SLNMM的检出使大肠癌的Dukes分期更加精细,从而有助于指导术后的辅助治疗和预后判断。

乳腺癌中VEGF-C mRNA的表达与淋巴结微转移的关系

目的探讨腋淋巴结阴性(axillary node negative,ANN)乳腺癌中VEGF-C mRNA的表达与淋巴结微转移的关系及临床意义。方法随机抽取乳腺浸润性导管癌139例,在组织芯片上用原位杂交方法检测VEGF-C mRNA表达;用免疫组化EnVision法检测广谱角蛋白(AE1/AE3)在ANN组淋巴结中的表达,以判定微转移;同时检测ER、ki67和cerbB-2蛋白在肿瘤组织中的表达情况。结果VEGF-C mRNA在腋淋巴结阳性(axillary node positive,ANP)组中的表达(53.0%)显著高于ANN组(27.4%)(P=0.001 1)。ANN组中,有14例患者腋窝淋巴结发现微转移灶,微转移率为19.2%(14/73);有微转移的患者其原发灶中VEGF-C mRNA表达率(50.0%)高于同组无微转移患者(22.4%)。差别具有统计学意义(P=0.038 6)。ANN组中VEGF-C mRNA表达与肿块大小(临床分期)、组织学分级正相关;与ER、ki67和cerbB-2蛋白表达及患者年龄无关。结论在ANN乳腺癌患者中VEGF-C mRNA的表达与淋巴结微转移密切相关。检测癌组织中VEGF-C mRNA的表达可能为预测ANN乳腺癌患者淋巴道转移和微转移提供有用的观测指标。

联合免疫组化和PCR检测CK20在早期宫颈鳞癌淋巴结中的表达和意义

目的:探讨早期宫颈鳞癌淋巴结微转移阳性率,阳性淋巴结分布以及与病理类型、临床分期之间的关系。方法:用免疫组化法和RT-PCR法分别及联合检测492枚(52例)早期宫颈鳞癌手术切除淋巴结中CK20的表达情况。结果:联合法检测CK20阳性率与单独使用一种方法的阳性率相比有显著性差异,且显著高于常规HE染色(P<0.05)。闭孔淋巴结CK20阳性率最高。CK20表达与宫颈鳞癌临床分期、病理分级有关(P<0.05),而与年龄无关(P>0.05)。结论:联合免疫组化和PCR检测CK20敏感性较高,闭孔淋巴结可作为检测CK20表达的首选淋巴结,对于评价早期宫颈癌微转移具有重要意义。

淋巴结微转移检测对结直肠癌病理分期的影响

目的：探讨淋巴结微转移对结直肠癌患者病理分期的影响。方法：对2003年3月至2008年10月间手术切除的60例结直肠癌患者的990枚淋巴结进行常规HE染色和CK20免疫组化检查,检测结直肠癌的淋巴结转移,并对结果进行统计学分析。结果：应用HE染色法淋巴结转移检出率27.2%（269/990）,CK20免疫组化淋巴转移检测率为37.2%（368/990）,两者差异有统计学意义（P〈0.05）。99枚淋巴结检出有微转移,11例TNM分期提高,HE染色重新分期率为18.3%（11/60）。结论：CK20免疫组化的检测可以显著提高淋巴结转移的检出率,有助于更准确地进行结直肠癌的临床病理分期。

pN0期非小细胞肺癌淋巴结微转移的临床分析

目的:研究pN0期非小细胞肺癌(non-small cell lung cancer,NSCLC)淋巴结微转移的临床病理特征,并分析微转移与患者生存率的关系。方法:收集89例行肺癌根治术并经病理检查证实为pN0期NSCLC患者的淋巴结石蜡切片,使用抗细胞角蛋白抗体检测微转移,分析淋巴结微转移的临床病理特点及其与患者生存率的关系。结果:pN0期NSCLC淋巴结微转移率为18.0%,其与肿瘤部位(P=0.030)、T分期(P=0.041)有关;N1区域淋巴结微转移度高于N2区域淋巴结(P=0.024),N2区域淋巴结跳跃性微转移率为5.62%;淋巴结微转移患者5年生存率低于无转移者(P=0.036)。生存分析显示淋巴结微转移是pN0期NSCLC的独立危险因素。结论:pN0期NSCLC的淋巴结微转移率较高,并可出现跳跃性微转移。术中应重视肺门、纵隔淋巴结清扫。淋巴结微转移预示pN0期NSCLC患者预后不良。

早期乳腺癌前哨淋巴结转移39例病理学研究

目的 探讨提高早期乳腺癌前哨淋巴结活检阳性率的方法。方法 回顾性分析 39例早期乳腺癌前哨淋巴结定位活检病例。对 2 8例阴性前哨淋巴结及所属非前哨淋巴结 ,在原组织蜡块平面 10 0 μm深处切片 2张 ,1张HE染色 ,1张免疫组化染色 ;于 2 0 0 μm深处切片 1张 ,行HE染色。 结果 39例中前哨淋巴结转移率是非前哨淋巴结的 4 3倍 ;多层切片、HE染色及免疫组化染色检测 2 8例 6 7枚阴性前哨淋巴结和 4 2 4枚非前哨淋巴结 ,2例 (2枚 )发现有转移 ,与单切面HE染色法相比 ,敏感性提高 7 1% ;前哨淋巴结假阴性率由 8 3%降为 7 1%。结论 前哨淋巴结是乳腺癌最早发生也是最易发生转移的淋巴结 。

口腔鳞癌颈淋巴结微转移的检测及临床意义

目的:探讨口腔鳞癌颈淋巴结微转移的发生率及临床意义。方法:对46例口腔鳞癌常规病理学检查阴性的颈淋巴结标本作连续切片,并采用免疫组化和流式细胞技术(FCM)检测微转移,以阳性淋巴结作为对照。随访5a。结果:46例口腔鳞癌患者的276枚常规病理检查阴性的颈淋巴结,免疫组化检测有5例8枚淋巴结有微转移(10.87%,5/46)。FCM检测200枚淋巴结,原8枚免疫组化检测阳性的淋巴结全部为阳性,阳性数为0.49%～6.06%。5例淋巴结微转移患者,4例复发。结论:免疫组化和流式细胞技术可提高口腔鳞癌颈淋巴结微转移发现的机率,对术后辅助治疗和预后评估具有一定的指导意义。

流式细胞术检测食管鳞癌淋巴结微转移的临床病理意义

目的:选定食管鳞癌特异性标记物CK-19用流式细胞术检测食管鳞癌区域淋巴结是否存在微转移,分析其与预后的关系,探寻"逆向预后"的原因,指导临床治疗。方法:CK-19基因表达于食管上皮组织,但在正常淋巴结不表达。1998年1月至1999年12月随机抽取根治性切除食管鳞癌100例,共清扫出淋巴结1248枚。用流式细胞术检测光镜下阴性的淋巴结分析影响淋巴结微转移的相关因素。结果:279枚常规病理学检测转移的淋巴结经流式检测证实,67例及969枚光镜阴性淋巴结,流式细胞术检测其中28例及156枚淋巴结有微小转移,其微小转移率为41.8%,微小转移度为16.1%,其1、3、5年生存率分别为82.1%、53.5%及14.3%。淋巴结微小转移与病变浸润深度及癌细胞分化程度有关,而与年龄、性别、病变部位、病变长度及分期无关。淋巴结微转移和预后密切相关。结论:食管癌区域淋巴结确实存在普通光镜难以发现的微小转移,部分淋巴结阴性患者生存期短预后差原因可能与该患者存在淋巴结微小转移有关即"逆向预后",因此对食管癌手术标本应常规检测其淋巴结微转移,对存在淋巴结微转移者,术后给予相应的辅助治疗。

前哨淋巴结活检及微转移检测对乳腺癌的临床诊疗价值

目的研究前哨淋巴结活检及微转移检测对乳腺癌的临床诊疗价值。方法选取乳腺癌患者133例,对其首先进行前哨淋巴结定位活检术,然后行根治性腋窝淋巴结清扫术。结果 133例乳腺癌患者中,每例检出前哨淋巴结1～5枚,平均3枚,共399枚;检出非前哨淋巴结5～20枚,平均(12. 5±2. 4)枚,共1596枚。单一层面切片的HE染色,前哨淋巴结发现45枚(8例)(17. 7%),非前哨淋巴结41枚(14例)(34. 1%)发现转移。对399枚前哨淋巴结行多层切片HE染色及免疫组化染色检测,发现单层切片常规HE染色前哨淋巴结均有微小转移,余下HE染色阴性的前哨淋巴结有微小转移灶,直径均小于2 mm,累计有96枚前哨淋巴结内有转移灶;对早期乳腺癌前哨淋巴结分别进行多层切片HE染色及免疫组化染色法与单一层面切片HE染色法相比,检出率提高,差异有统计学意义(P <0. 05)。结论前哨淋巴结活检及微转移的检测结果可以用来指导是否对乳腺癌患者进行常规腋淋巴结清扫,对其进行连续切片及免疫组化染色检测比普通HE染色检出率明显提高。

病检阴性胃癌淋巴结转移的检测:免疫组化和RT-PCR的比较

目的:比较免疫组化和RT-PCR方法对常规病检阴性(pN0)胃癌淋巴结微转移的检测价值。方法:研究pN0胃癌14例249枚淋巴结,分别用癌胚抗原和CK19作为标记物,用免疫组化和RT-PCR方法检测淋巴结微转移。结果:在6例患者的淋巴结中发现微转移,免疫组化和RT-PCR法分别发现11枚和19枚微转移淋巴结,2种方法发现的微转移阳性率分别为4.4%和7.6%。在免疫组化检测阳性的11枚淋巴结中,8枚用CK19和癌胚抗原的抗体染色均为阳性,另3枚仅在CK19抗体染色时阳性而癌胚抗原的抗体染色阴性。在19枚RT-CPR检测阳性的淋巴结中,15枚用CK19和癌胚抗原作为标记物检测均为阳性,另4枚仅在用CK19检测阳性而用癌胚抗原作为标记物检测为阴性。所有用免疫组化检测为阳性的淋巴结用RT-PCR检测均为阳性,但19枚RT-PCR检测阳性的淋巴结中枚有10枚被免疫组化证实为阳性。结论:RT-PCR是比免疫组化更为敏感的检测胃癌淋巴结微转移的方法,而CK19作为检测标记物比癌胚抗原更敏感。

前哨淋巴结活检在早期胃癌中的应用及微转移检测

目的:探讨前哨淋巴结活检(SLNB)技术在早期胃癌中的应用及前哨淋巴结(SLN)微转移检测的临床意义。方法:34例早期胃癌术中亚甲蓝定位活检前哨淋巴结,术后行常规HE病理和角蛋白单克隆抗体AE1/AE3免疫组化(ELIVISION二步法)检查。结果:34例中33例检出SLN,检出率为97.1%。由SLN状态预测胃周淋巴结转移情况的准确率为93.9%(31/33),敏感性为88.9%(16/18),特异性为100.0%(16/16),假阴性率为11.1%(2/18)。免疫组化法与常规HE法对SLN转移的检出率相比较,差异有统计学意义(P=0.034),免疫组化法共检出12枚/8例SLN存在微转移现象。结论:SLN能较准确反映早期胃癌的淋巴结转移状况,免疫组化法较常规HE病理检查更为敏感,AE1/AE3免疫组化法能提高SLN微转移的检测,对确定临床分期、诊疗及判断预后有积极临床意义。

临床N0舌鳞癌哨位淋巴结微转移检测及其意义

目的：以CK19mRNA为标志物，采用RT—PCR法检测临床NO舌鳞癌哨位淋巴结（sentinel node。SN）的微转移状况，从基因水平探讨择区颈清扫的必要性。方法：采用1探头示踪法对21例临床NO舌鳞癌患者进行SN示踪，采用RT—PCR法检测CK19基因在SN中的表达，并与颈部慢性炎症淋巴结和舌鳞癌转移淋巴结各10枚的CK19基因表达进行对比。采用SPSS10．0软件包进行X^2检验。结果：21例临床N0舌鳞癌中，20例SN示踪成功，共取得SN53枚，每例1～4枚，平均2．65枚。RT—PCR法检测CK19基因表达诊断颈部淋巴结转移的敏感度为90％，特异度为80％。SN示踪成功的20例中，常规病理检查发现淋巴结转移2例，隐匿性转移率为10％；51枚常规病理阴性的SN中，CK19-mRNA阳性表达11枚（来自7例患者），微转移发生率为21．57％，隐匿性转移率为35％，2种方法评价舌鳞癌隐匿性颈淋巴结转移率有显著性差异，P=0．013。结论：由于微转移的存在，以SN常规病理分析来决定颈清扫术式并不可靠，择区颈清扫仍然是临床NO舌鳞癌颈部治疗的首选方法。

早期宫颈癌淋巴结微转移的临床研究

目的探讨早期宫颈癌淋巴结内细胞角蛋白(CK)和鳞状细胞癌抗原(SCCA)表达的临床意义。方法将39例宫颈鳞癌患者的269枚无癌转移的淋巴结行免疫组化染色检测CK、SCCA,分析微转移与临床预后相关因素的关系。结果在原发病灶中CK和SCCA均呈阳性表达(100.00%)。在所有淋巴结中,CK的阳性率为3.35%(9/269),SCCA的阳性率为1.49%(4/269)。随访发现,肿瘤的复发/转移情况与淋巴结的微转移状态存在明显相关(P=0.002)。结论早期宫颈癌患者存在淋巴结微转移风险,淋巴结的微转移状态与术后复发/转移情况密切相关。

食管癌淋巴结微转移检查

[目的]通过食管癌淋巴结微转移的检测,探讨食管癌淋巴结转移规律。[方法]对某医院2002～2004年间50例pN0食管鳞癌病人的淋巴结病理切片150张,根据淋巴结原发部位进行分组,单抗CK5&6免疫组化染色,比较不同部位淋巴结微转移的差异。[结果]微转移阳性检测率达12%,病灶旁淋巴结微转移阳性率最高,分别为21.7%和20%。左喉返神经旁淋巴结16.7%,胃左及小网膜淋巴结10.3%,隆突下淋巴结的阳性率最低,2%。[结论]胸段食管癌淋巴结微转移最常见的部位是病灶旁和左喉返神经旁,食管癌手术中应注意上述部位淋巴结的清扫。