Immunohistochemical detection of lymph node microinvolvement in node-negative gastric cancer

Background. Despite curative resection of the primary tumor with extensive dissection of lymph nodes, some patients with node-negative gastric cancer die of local relapse or metastatic disease. Microinvolvement of regional lymph nodes may play an important role in the prognosis. Methods. To evaluate the incidence and prognostic implications of regional lymph node microinvolvement in node-negative gastric cancer, we retrospectively analyzed tissue samples from 51 patients operated on for primary gastric cancer. They had tumors that invaded beyond the muscularis propria, but without metastasis to the lymph nodes, shown by the conventional H&E staining method. The regional lymph nodes were examined immunohistochemically, using monoclonal antibodies against cytokeratin. Results. Microinvolvement was found in 4.8% of lymph nodes (67/1390) and in 43.2% of the patients (22/51). The clinical outcome of the patients with microinvolvement was not significantly different from those without it. However, no patient died in the no-microinvolvement group, while three patients in the microinvolvement group died of recurrence. Conclusion. The incidence of microinvolvement in conventionally negative lymph nodes cannot be ignored, and detecting microinvolvement may be important for predicting recurrence of gastric cancer. Received for publication on April 26, 1999; accepted on Sept. 14, 1999     

Detection of sentinel lymph nodes in gastric cancers based on immunohistochemical analysis of micrometastases

BACKGROUND AND OBJECTIVES: The sentinel lymph node (SN) theory has the potential to change the trend of surgery for gastric cancer that is based on wide resection of the stomach with dissection of regional lymph nodes. However, feasibility tests of SN mapping procedures in gastric cancers with analysis of micrometastasis are rare. This study aimed to estimate the clinical usefulness of SN mapping using a dual procedure with dye- and gamma probe-guided techniques for gastric cancers, based on immunohistochemical staining (IHC) analysis. METHODS: SN mapping procedures were performed on 41 patients with T1-T2 gastric cancer, and gastrectomy with D2 lymphadenectomy followed. All SNs and non-SNs obtained from the patients were tested by IHC analysis using anti-cytokeratin antibodies. RESULTS: Using the dual mapping procedure, SNs were detected in all patients (100%). SN was positive in all patients with lymph node metastasis except in one with non-solid type poorly differentiated adenocarcinoma with marked lymphatic permeation, thus achieving an accuracy rate of 98%. CONCLUSIONS: The method was accurate in predicting nodal status and could be an indicator for less invasive treatment in patients with gastric cancer.     

The significances of lymph node micrometastasis and its correlation with E-cadherin expression in pT1-T3N0 gastric adenocarcinoma

BACKGROUND: Recently, micrometastasis (MM) in the lymph node in gastric cancer has been detected by cytokeratin immunostaining. However, clinical significance of MM and its relationship with reduced expression of E-cadherin in primary lesion have not been well studied. METHODS: The 4,990 lymph nodes from 184 pT1~T3N0 patients from 1995 to 2000 at Korea University Hospital were immunostained with the anticytokeratin AE1/3 antibody for detection of micometastasis. The primary lesions were also immunostained for E-cadherin expression. RESULTS: MM in the lymph node of gastric cancer was found in 131 (2.6%) of total dissected nodes, and 31 of 184 patients (16.8%) were shown to have MM. The MM was significantly correlated with the depth of invasion, tumor size, operation method, Lauren classification, lymphovascular invasion and loss of E-cadherin expression in primary tumor. On multivariate analysis, the independent risk factors for MM were the depth of invasion and loss of E-cadherin expression. The patients with MM had significantly lower 5-year overall and disease free survival rate than those without MM. CONCLUSION: Lymph node MM in histologically node-negative gastric cancer was significantly correlated with poor 5-year survival rate. The determination of E-cadherin expression in primary gastric tumor may be useful in prediction of the MM.     

Paraaortic lymph node micrometastasis and tumor cell microinvolvement in advanced gastric carcinoma

Background. Paraaortic lymph node dissection in advanced gastric carcinoma is controversial. The purpose of this study was to investigate the incidence and significance of micrometastasis (MM) or tumor cell microinvolvement (TCM) in these critical lymph nodes. Methods. A total of 2339 lymph nodes, including 390 paraaortic nodes, obtained from 47 patients with advanced gastric carcinoma were examined immunohistochemically, using cytokeratin antibody. Results. Lymph node metastasis was found in 95 of the 390 paraaortic nodes of 14 patients by routine histological examination. MM or TCM was immunohistochemically detected in 45 of the 295 negative paraaortic lymph nodes from 15 of 33 patients (MM, n = 5; TCM, n = 10). The 5-year-survival rate in the paraaortic node-negative group and cytokeratin-positive group was significantly higher that that of the hematoxilin and eosin-positive group. The total number of lymph node metastases by hematoxylin and eosin staining and the pathological lymph node compartments, by cytokeratin-positive nodes, were prognostic factors by multivariate analysis. Conclusions. We demonstrated a high rate of MM or TCM in the paraaortic lymph nodes and suggest that such harbored metastases are related to the prognosis of patients with advanced gastric carcinoma. On the basis of this study, a multi-institutional study should be considered. Received for publication on June 7, 1999; accepted on Sept. 30, 1999     

N0期胃癌患者的临床病理特征及预后分析

目的:探讨N0期胃癌患者的临床病理特征及影响预后的危险因素。方法:回顾性分析2003年3月至2012年3月我科收治的296例N0期胃癌患者的临床病理资料。分析其临床病理特征以及通过单因素和多因素分析影响其预后的危险因素。结果:本研究中男性248例,女性48例。中位年龄58岁（28~82岁）。中位肿瘤大小4 cm（0.3~15 cm）。168例患者为TNM I期,126例患者为TNM II期,仅2例患者为TNM III期。患者的中位随访时间为62.3个月（1~73.4个月）。1、3、5年总体生存率分别为97.6%、89.2%和83.7%。单因素分析显示切除方式、CEA、CA125、脉管侵犯、T分期和TNM分期为影响N0期胃癌患者预后的危险因素（P<0.05）。然而,仅CEA、CA125、脉管侵犯和T分期为影响预后的独立危险因素（P＜0.05）。结论:N0期胃癌患者总体预后较好。CEA、CA125、脉管侵犯和T分期为影响N0期胃癌患者预后的独立危险因素。     

Comparison of monoclonal antibodies for the detection of occult breast carcinoma metastases in bone marrow

Summary Twenty percent (n = 6) of Stage III or IV breast cancer patients (n = 30) had bone marrow metastases detected in bilateral bone marrow biopsy/aspiration preparations using standard histologic preparations. Each metastasis was also detected by four separate monoclonal antibodies (MAbs) which recognize breast carcinoma associated antigens (DF3, anti-EMA, HMFG-2, and CAM5.2). These MAbs were then utilized to stain other bone marrow preparations (n = 81) to determine their utility for the detection of micrometastatic breast carcinoma. MAbs HMFG-2, anti-EMA, and DF3 were each strongly reactive with bone marrows containing histologically-evident metastatic breast carcinoma (18/18). These anti-epithelial membrane antigen MAbs, however, were also reactive with rare plasma cells and immature cells (as well as cell clusters) in some of the control bone marrow samples tested, including those from normal patients and patients with hematologic disorders. They also reacted with some of the preparations from patients with leukemia and lymphoma, and with uninvolved marrows from patients with non-epithelial malignancies. The anti-keratin MAb CAM5.2, in contrast, reacted with 83% (15/18) breast cancer metastases and failed to stain any cells in the various categories of control marrow preparations. These data suggested that MAb CAM5.2 might be utilized to immunohistochemically differentiate micrometastatic breast carcinoma from immature myeloid or erythroid elements.Each MAb was then reacted with histologically uninvolved marrow preparations from the remaining 24 of 30 breast cancer patients in an attempt to identify occult breast carcinoma metastases. While MAbs HMFG-2, DF3, and anti-EMA demonstrated reactive cells in some of these marrows, this reactivity was similar to that seen with control preparations. MAb CAM5.2, in contrast, was negative with all specimens. These data suggest that MAb CAM5.2 may be a useful immunologic probe for the detection and confirmation of metastatic breast carcinoma in bone marrow, while more caution must be employed in the interpretation of results obtained using MAbs anti-EMA, DF3, and HMFG-2.     

早期胃癌淋巴结微转移检测的临床意义

目的探讨淋巴结微转移与早期胃癌临床病理特征及预后的关系。方法应用免疫组化方法对182例早期胃癌1631个淋巴结进行微转移检测。结果早期胃癌中4.1%的淋巴结、13.2%的患者存在淋巴结微转移。淋巴结微转移与年龄、性别、肿瘤大小、部位、大体类型、分化程度、浸润深度、淋巴管癌栓、癌旁黏膜萎缩、肠化、异型增生及预后无关。结论淋巴结微转移检测对早期胃癌预后判断价值有限。     

Risk factors for nodal micrometastasis of submucosal gastric carcinoma: assessment of indications for endoscopic treatment

Background. Despite the widespread use of endoscopic mucosal resection (EMR) for intramucosal gastric carcinoma, there is no standardized therapy for those patients in whom the carcinoma is found, after EMR, to have invaded the submucosa. Our aim in this study was to examine the relationship between the clinicopathological features of submucosal invasive carcinomas and their incidence of nodal micrometastasis, as detected by anti-human cytokeratin immunohistochemistry, in order to assess the curative potential of submucosal carcinoma by EMR. Methods. Fifty surgically resected submucosal gastric carcinomas which would have satisfied the absolute indications for EMR, except for the criterion of submucosal invasion, were examined. The extent of submucosal invasion was determined by measuring its vertical and horizontal spread. Immunohistochemical analysis was performed with anti-human cytokeratin antibody (CAM5.2). Results. Three of 50 cases (6.0%) were positive for nodal metastasis by routine H&E examination. Nodal micrometastases were detected in 11 of 47 cases (23.4%). Statistical analysis revealed that both the depth and the width of carcinoma in the submucosa were significantly larger in cases with than in those without micrometastasis (P = 0.019 and P = 0.006, respectively). The group with lymphatic invasion showed more frequent micrometastasis than the group without (P = 0.014). There were no micrometastases in submucosal carcinomas whose submucosal invasion was less than 200 μm vertically and less than 320 μm horizontally. Conclusions. The present study indicates that differentiated gastric adenocarcinoma with minimal submucosal invasion (less than 200 μm vertically and less than 320 μm horizontally) and not accompanied by peptic ulcer or other risk factors, such as lymphatic invasion, can be considered as having high curative potential by EMR alone, without the necessity for further radical surgery. Received for publication on Nov. 6, 1998; accepted on Feb. 19, 1999     

Immunohistochemical staining of bone marrow biopsies for detection of occult metastasis in breast cancer

Summary Immunohistochemical (IHC) techniques should allow for a greater detection of bone marrow micrometastasis in patients with breast carcinoma. We studied a series of bone marrow (BM) biopsies negative by conventional histologic techniques from 93 patients with breast carcinoma. Prior to this study, twelve BM biopsies, positive by conventional histology, were stained with a panel of monoclonal antibodies (MoAb), directed either against cytokeratin (KL1, AE1-AE3, CAM5-2) or epithelial membrane antigen (EMA, HMFG2). KL1 appeared to be the most sensitive of the markers used in the detection of metastases and is available commercially. It therefore was the only MoAb used with the series of 93 BM biopsies negative by conventional examination. Within this series, among 45 patients clinically suspected of having bone marrow metastasis but with BM biopsies negative by conventional staining, one case showing myelofibrosis stained positive with KL1 demonstrating isolated tumor cells. For the 48 patients without suspicion of bone marrow metastasis at initial diagnosis for breast carcinoma, KL1 revealed no marrow metastasis.Single bone marrow biopsy techniques whether stained by conventional or IHC methods do not appear to be useful tests to detect occult bone marrow metastasis, especially at initial diagnosis of clinically M_o breast carcinoma patients.     

Quantitative detection of micrometastases in the lymph nodes of gastric cancer patients with real-time RT-PCR: a comparative study with immunohistochemistry

Histologic examination lacks the sensitivity to detect micrometastases in gastric cancer lymph nodes. In the present study, we applied a real-time RT-PCR approach to the quantitative detection of micrometastases in gastric cancer lymph nodes and compared diagnostic power with routine histology and immunohistochemistry. We studied 392 lymph nodes from 21 gastric cancer patients who underwent curative surgery. Real-time quantitative RT-PCR was performed on a LightCycler instrument using a hybridization probe for carcinoembryonic antigen (CEA) and cytokeratin-20 (CK20) as marker genes. Immunohistochemistry with antibodies to wide-keratin was also performed in the lymph nodes to compare the sensitivity and specificity. Median (average) values of CEA mRNA in lymph nodes in patients with histology(+), immunohistochemistry(+)/histology(-), immunohistochemistry(-)/histology(-) and negative control results were 4600 (16000), 200 (400), 0 (9.8) and 0 (0.6), respectively. There were some false-negative results with simple CEA and CK20 real-time RT-PCR due to the presence of low gene-expressing gastric cancers as revealed by CEA and CK20 immunohistochemistry. CEA in combination with CK20 (duplex) real-time RT-PCR partially covered this weakness. Consequently, all 71 histology(+) lymph nodes were positive for duplex real-time RT-PCR as well as wide-keratin immunohistochemistry. Positivity rates by histology, wide-keratin immunohistochemistry and duplex real-time RT-PCR were 18.0% (71/392), 20.9% (82/392) and 25.8% (101/392), respectively. In 2 of 8 patients with pT1N0, positive lymph nodes were observed by real-time RT-PCR but not by immunohistochemistry. These results indicate that duplex quantitative real-time RT-PCR is the most sensitive method for detecting micrometastases and useful for evaluating the prognostic significance of lymph node micrometastasis in gastric cancer patients.     

Clinical significance of micrometastasis in bone marrow of patients with gastric cancer and its relation to angiogenesis

Background. Detection of micrometastasis is an important problem of clinical significance for a better understanding and control of tumor progression, which will improve patients' survival time. Methods. To identify micrometastases in bone marrow, an immunocytochemical assay for epithelial cytokeratin protein was performed in 106 patients with primary gastric cancer. Also, in 40 of the 106 patients, vascular endothelial growth factor (VEGF) expression and intratumoral vessel density were examined by an immunohistochemical staining method. Results. Of the 106 patients, 22 (20.8%) presented with cytokeratin-positive cells in bone marrow at the time of primary surgery. The positive findings were related to depth of invasion, peritoneal dissemination, and liver metastasis. Patients with cytokeratin-positivity in bone marrow had a higher VEGF positive rate (73%; 8/11) than did cytokeratin-negative patients (48%; 14/29). Intratumoral vessel density in VEGF-positive patients was 26.9 ± 10.3, which was significantly higher than that in VEGF-negative patients (13.2 ± 8.7, P < 0.05). Thus, the presence of cytokeratin-positive cells in bone marrow was closely related to angiogenesis in the primary tumor. Conclusions. Cytokeratin staining can be useful for identifying patients at high risk for metastasis. Prophylactic lymph node dissection, adjuvant chemotherapy, and antiangiogenic treatment may be necessary for patients with micrometastasis. Received for publication on Jan. 27, 1999; accepted on Feb. 26, 1999     

微小胃癌、小胃癌31例临床病理分析

目的探讨微小胃癌和小胃癌临床、独特的生长模式及病理组织学变迁的特点,利于选择治疗方案。方法对31例微小胃癌和小胃癌患者临床及病理资料回顾性分析。结果微小胃癌及小胃癌组占同期早期胃癌病例的10.6%;微小胃癌组、小胃癌组高、中分化型分别为72.7%、55.0%;黏膜内癌分别为81.8%、35.0%;微小胃癌组不伴淋巴结转移,小胃癌组淋巴结转移率为5%;两组脉管浸润均阴性;微小胃癌组、小胃癌组癌肿表层部与浸润部病理组织学同型性分别为90.1%、85.0%;微小胃癌组及小胃癌组浸润部较表层部分化程度趋向低下者9.7%,趋向高分化3.2%。结论微小胃癌组较小胃癌组高、中分化型比例高、浸润浅;小胃癌可出现淋巴结转移;随着肿瘤的发展,微小胃癌及小胃癌自表层部向浸润部发展,病理组织学多趋低分化变迁;绝大部分微小胃癌及半数以上（高、中分化型、无淋巴结转移、无脉管浸润者）的小胃癌,可选择内镜下微创治疗。     

胃癌组织UHRF1表达与预后相关性分析

目的探讨泛素样含PHD和环指域1(ubiquitinlike with PHD and ring finger domain 1,UHRF1)表达在胃癌发生发展过程中的作用及其预测胃癌患者预后的价值。方法采用免疫组织化学法检测2002-02-01-2004-05-31手术切除106例胃癌组织和72例正常胃黏膜组织中UHRF1的表达情况,统计分析其表达与临床病理特征及预后的关系。结果胃癌组织中UHRF1阳性表达率为84.0%(89/106),癌旁正常胃黏膜组织阳性率为48.6%(35/72),差异有统计学意义,χ2=35.220,P<0.001。UHRF1表达与胃癌分化程度(P<0.05)、TNM分期(P<0.01)、是否淋巴结转移(P<0.05)和是否远处转移(P<0.05)密切相关,而与年龄(P=0.351)、性别(P=0.427)无关。单因素和多因素分析结果均提示,UHRF1高表达患者生存时间短于低表达患者,P<0.001。结论 UHRF1与胃癌的发生发展密切相关,其表达强度可以作为患者预后的重要指标。     

细胞角蛋白在肿瘤性滋养细胞的检测中的应用价值

应用免疫组化方法，采用广谱抗细胞角蛋白单克隆抗体，检测１２９例滋养细胞肿瘤（葡萄胎３０例，侵蚀性葡萄胎４１例，绒毛膜癌５８例）中肿瘤性滋养细胞内细胞角蛋白表达。结果表明，肿瘤性合体滋养细胞（ＳＴ）、中间型滋养细胞（ＩＴ）及细胞滋养细胞（ＣＴ）内细胞角蛋白的阳性表达率均为１００％。ＳＴ呈强阳性表达，其表达形式为胞浆型；ＩＴ呈中－强阳性表达，多数为胞浆型；ＣＴ呈中等强度阳性，一般为质膜型。我们认为，细胞角蛋白实为肿瘤性滋养细胞（包括转移瘤）的一种更为敏感、可靠的标志物，在滋养细胞肿瘤的病理诊断、鉴别诊断及肿瘤浸润范围的确定方面具有重要的应用价值。     

胃癌腹膜微转移检测及其与胃癌预后关系的研究进展

胃癌的腹膜转移和复发是影响胃癌患者预后的重要因素,早期诊断腹膜微转移对提高患者的生存及预后有重要意义。应用灵敏度高的检测方法,选择特异性高的标志物能提高胃癌腹膜微转移的检测率,在肿瘤的诊治以及提高胃癌患者的生存率提供帮助。     

胃癌腹膜微转移检测及其与胃癌预后关系的研究进展

胃癌的腹膜转移和复发是影响胃癌患者预后的重要因素,早期诊断腹膜微转移对提高患者的生存及预后有重要意义.应用灵敏度高的检测方法,选择特异性高的标志物能提高胃癌腹膜微转移的检测率,在肿瘤的诊治以及提高胃癌患者的生存率提供帮助.     

Clinicopathological significance of c-erbB-2 protein expression in human gastric carcinoma.

One hundred seventy-nine primary human gastric tumors not associated with early cancer or noncurative resection were examined immunohistochemically for the expression of c-erbB-2 protein. Positive staining, regarded as an indication of gene amplification, was evident in 22(12%) of the tumors. Of various clinicopathological factors considered, a statistically significant difference in association with frequency of expression was noted only for histological differentiation, as follows: 39% positive staining in papillary, 17% in well differentiated, 5% in moderately differentiated, and 4% in undifferentiated adenocarcinomas (P greater than 0.01). The 5-year survival rates of patients with positive and negative c-erbB-2 staining were 57% and 59%, respectively. These findings indicate that, in the case of human gastric adenocarcinoma, expression of c-erbB-2 protein is correlated with tumor histological differentiation. Our results also suggest that the presence or absence of c-erbB-2 protein may not serve as a prognostic indicator, particularly in cases of adenocarcinoma of the stomach.     

78例乳腺癌改良根治术后pT3N0M0期患者放疗价值探讨

目的 探讨乳腺癌改良根治术后病理分期为T₃N₀期患者的术后放疗价值。方法 回顾分析1997-2014年收治的乳腺癌改良根治术后患者资料,筛选标准为女性、术后病理提示浸润性癌、肿瘤最大径>5 cm且腋窝淋巴结未见转移、未接受新辅助化疗及内分泌治疗,且无远处转移及其他第二原发癌。78例符合条件。40例(51%)接受术后放疗,67例(86%)接受辅助化疗。Kaplan-Meier法计算DFS、OS及LRR率,组间差异用Logrank法检验。结果 中位随访时间79个月(6~232个月),5年OS、DFS和LRR分别为89%、87%和2%。放疗组与未放疗组患者5年DFS分别为84%与91%(P=0.641),5年OS分别为84%与96%(P=0.126),5年LRR分别为0%和5%。仅ER/PR状态、分子分型影响患者DFS (P=0.002、0.031)。未放疗组有1例患者出现胸壁复发。结论 乳腺癌改良根治术后T₃N₀M₀期患者LRR率较低,仅ER/PR状态及分子分型影响患者DFS。在有效系统全身治疗基础上术后病理T₃N₀患者可能不需全部接受胸壁+锁骨上野放疗,但仍需大样本病例证实。