Altered expression of sialyl-Tn,Lewis antigens and carcinoembryonic antigen between primary and metastatic lesions of uterine cervical cancers

Summary Immunohistochemical examination was performed of serial sections of 24 normal human adult cervical tissues and 53 human cervical carcinomas including 36 cases with lymph node metastasis. For this investigation, monoclonal antibodies directed to Lewis-X, Lewis-Y, sialyl-dimeric Lewis-X (SDLX), sialyl-Tn (STn) and carcinoembryonic antigen (CEA) were used. STn and CEA antigens were expressed very weakly in the normal cervical epithelium but strongly in the cancer cells, indicating the antigens to be oncogenic antigens of cervical squamous cell carcinoma. No significant difference in immunoreactivity was observed between primary and metastatic lesions of carcinoma or between primary lesions with and without metastasis. However, the expression patterns of STn and Lewis-Y antigens were quite different between primary lesions and metastatic lesions. In primary lesions the cancer cell nests tended to be stained centrally, but in metastatic lesions the cancer cell nests tended to be stained peripherally. This finding may reflect an important role of these carbohydrate chains in the process of metastasis of cervical squamous cell carcinoma to regional lymph nodes.     

肺癌组织中岩藻糖化糖链结构免疫组化研究

Lewis X(Le~x)、唾液酸化的Lewis X(Sialyl Lewis X,SLe~x)和唾液酸化的双岩藻糖Lewis X(Sialyl Dimeric Lewis X,SDLe~x)是细胞表面外侧带α1,3岩藻糖的糖链结构。本文用免疫组化ABC法研究了肺癌原发灶、转移灶和癌旁组织中这三种抗原结构的表达。结果发现这三种抗原在肺癌细胞表面及胞浆中均有不同程度的表达,而在肺癌癌旁组织及正常肺组织中未见表达。有转移的肺癌和(或)低分化肺癌中这三种抗原结构的表达要明显高于未发生转移和高、中分化肺癌中相同抗原结构的表达。其中以SLe~x的表达与肺癌细胞的转移能力和分化程度关系最为密切。另外,肺癌浸润转移的淋巴结中也有Le~x、SLe~x的明显表达和SDLe~x的少量表达,而未被肺癌浸润转移的淋巴结中就没有它们的表达。     

宫颈鳞状细胞癌中ADAM17 的表达及其临床意义

目的:探讨去整合素-金属蛋白酶17(ADAM17)在宫颈鳞状细胞癌中的表达及其临床病理意义。方法:运用免疫组织化学法分别检测正常宫颈上皮、宫颈鳞状细胞癌及宫颈上皮内瘤样变组织中ADAM17的表达,并分析其与宫颈癌临床分期及淋巴结转移的相关性。结果:ADAM17在正常宫颈上皮组织切片中无明显表达,在宫颈上皮内瘤样变组织中少部分表达,呈浅黄色,在宫颈鳞状细胞癌中癌细胞大量表达,ADAM17表达呈棕褐色,数量较多且浓染。不同临床分期的宫颈鳞状细胞癌组织中ADAM17的阳性表达率比较存在显著统计学差异(P0.05),且随临床分期的上升,ADAM17的表达逐渐升高;有淋巴结转移的宫颈鳞状细胞癌组织中ADAM17的阳性表达率显著高于无淋巴结转移的宫颈鳞状细胞癌组织,差异具有统计学意义(P0.05)。结论:ADAM17蛋白在宫颈鳞状细胞癌组织中呈异常高表达,且与宫颈鳞状细胞癌的临床分期和淋巴结转移密切相关,通过检测ADAM17蛋白的表达可能有助于宫颈鳞状细胞癌的诊断、治疗和预后预测。     

Immunohistochemical study of the expression of fucosylated glycoconjugates in human lung carcinoma]

Lewis X, Sialyl Lewis X and Sialyl Dimeric Lewis X are three fucosylated glycoconjugates on cell surface. With immunohistochemical method, the expression of the three structures in the original lung cancer tissues (with or without metastasis), adjacent tissues and metastatic lesions of lung carcinoma were studied. It was found that the three antigens were expressed with different intensity on the cell surface and in the cytoplasm of the lung carcinoma cell. However, there was no or only trace expression of these antigens in the adjacent tissues of lung carcinoma and normal lung tissues. Moreover, the original lesions of lung carcinoma with metastasis and/or poor differentiation expressed higher level of the three antigens than those without metastasis and/or with well or medium differentiation. Sialyl Lewis X was considered to be more closely related to the metastatic ability and differentiation of the lung carcinoma cell than the other two antigens, Lewis X and Sialyl Dimeric Lewis X. Furthermore, in the lymph nodes with lung carcinoma cell metastasis, there were expression of the three antigens with different degree, while in those lymph nodes without lung carcinoma cell metastasis, there was no expression of the three antigens.     

Establishment of a Patient-Derived Orthotopic Xenograft (PDOX) Model of HER-2-Positive Cervical Cancer Expressing the Clinical Metastatic Pattern

Squamous cell carcinoma of the cervix, highly prevalent in the developing world, is often metastatic and treatment resistant with no standard treatment protocol. Our laboratory pioneered the patient-derived orthotopic xenograft (PDOX) nude mouse model with the technique of surgical orthotopic implantation (SOI). Unlike subcutaneous transplant patient-derived xenograft (PDX) models, PDOX models metastasize. Most importantly, the metastasis pattern correlates to the patient. In the present report, we describe the development of a PDOX model of HER-2-positive cervical cancer. Metastasis after SOI in nude mice included peritoneal dissemination, liver metastasis, lung metastasis as well as lymph node metastasis reflecting the metastatic pattern in the donor patient. Metastasis was detected in 4 of 6 nude mice with primary tumors. Primary tumors and metastases in the nude mice had histological structures similar to the original tumor and were stained by an anti-HER-2 antibody in the same pattern as the patient’s cancer. The metastatic pattern, histology and HER-2 tumor expression of the patient were thus preserved in the PDOX model. In contrast, subcutaneous transplantation of the patient’s cervical tumors resulted in primary growth but not metastasis.     

CERNA2: A predictor for clinical progression and poor prognosis in cervical carcinoma

Competing long noncoding RNA 2 (lncRNA 2) for microRNA let-7b (CERNA2) has emerged as an important regulator of tumorigenesis and cancer progression but the clinical value and regulatory function of CERNA2 is yet to be investigated in cervical carcinoma. In our study, we found the CERNA2 expression was obviously increased in cervical carcinoma tissues compared with adjacent normal cervical tissues. In addition, we observed that metastatic lymph nodes exhibited high levels of CERNA2 expression in contrast to primary cervical carcinoma tissues. Furthermore, high CERNA2 expression was associated with advanced clinical stage, lymph node metastasis, distant metastasis poor histological grade, and short overall survival in cervical carcinoma patients. Moreover, high CERNA2 expression acted as an independent unfavorable predictor for overall survival in cervical carcinoma patients. The cell migration and invasion assays in vitro suggested that knockdown of CERNA2 remarkably inhibited cell migration and invasion in cervical carcinoma. In conclusion, CERNA2 functions as an oncogenic lncRNA and may be as a potential therapeutic target in cervical carcinoma.     

Cytologic features of metastatic papillary thyroid carcinoma in cervical lymph nodes

OBJECTIVE: To elucidate the cytologic characteristics of metastatic papillary thyroid carcinoma (PTC) in cervical lymph nodes and the differences in cervical lymph nodes from those of stage I (intrathyroidal) PTC. STUDY DESIGN: Forty-seven cases of papillary thyroid carcinoma with cervical lymph node metastasis (group A) and 38 cases of intrathyroidal papillary carcinoma (group B) were included in this study. Preoperative fine needle aspiration cytology (FNAC) examination was performed on enlarged cervical lymph nodes (47 cases, group A) and enlarged thyroid nodules (13 cases, group A, and 38 cases, group B). All the cases were surgically excised and pathologically verified. The cytologic smears were reviewed and analyzed. RESULTS: The cytologic characteristics of metastatic PTC in cervical lymph nodes displayed a higher frequency of foamy macrophages (51.1% vs. 26.3%) and a lower frequency of distinct cell borders (38.3% vs. 71.1%) than those of stage I PTC. Metastatic PTC in cervical lymph nodes also had a higher frequency of cystic degeneration (44.7% vs. 5.3%) than intrathyroidal lesions. In 1 of the 47 cases with lymph node metastasis, the aspirate contained macrophages but no tumor cells. CONCLUSION: FNAC was useful in the diagnosis of metastatic PTC in cervical lymph nodes. However, because cystic degeneration appeared frequently, FNAC combined with thyroid ultrasonography to find the primary lesion is necessary in this situation.     

PDGF—D和VEGF在宫颈鳞癌中的表达及其意义

目的：检测宫颈鳞癌组织中血小板源性生长因子D（PDGF—D）和血管内皮生长因子（VEGF）的表达,探讨二者在宫颈癌变过程中的作用及意义,为探讨宫颈鳞癌的发病机制和宫颈鳞癌的早期治疗提供理论依据。方法：采用免疫组织化学（sP法）检测40例宫颈鳞癌和10例正常宫颈组织中PDGF—D和VEGF蛋白的表达,分析两者之间的相关性及其与临床病理特征之间的关系。结果：宫颈鳞癌组织中PDGF—D和VEGF蛋白的表达显著高于正常宫颈组织（P〈0．05）;PDGF—D和VEGF的表达与宫颈鳞癌的分化程度及淋巴结转移有关（P〈0．05）;与年龄及临床分期无关（P〉0．05）。Spearman相关分析发现PDGF—D与VEGF表达程度呈正相关（r=0．346,P〈0．05）。结论：1．PDGF—D和VEGF在宫颈鳞癌组织中特异性高表达,可能在宫颈鳞癌的发生、发展与转移中起着重要作用。2．PDGF-D和VEGF表达与宫颈鳞癌的分化程度及淋巴结转移有关,与年龄及临床分期无关,提示它们可能在宫颈鳞癌的浸润和转移及预后方面有重要的监测意义。3．PDGF-D和VEGF在宫颈鳞癌组织中的表达呈正相关,提示两者起着相互促进的作用,对PDGF—D和VEGF的联合检查,为临床实际应用提供了参考。     

Histone Deacetylase (HDAC) 10 Suppresses Cervical Cancer Metastasis through Inhibition of Matrix Metalloproteinase (MMP) 2 and 9 Expression

Aberrant expression of histone deacetylases (HDACs) is associated with carcinogenesis. Some HDAC inhibitors are widely considered as promising anticancer therapeutics. A major obstacle for development of HDAC inhibitors as highly safe and effective anticancer therapeutics is that our current knowledge on the contributions of different HDACs in various cancer types remains scant. Here we report that the expression level of HDAC10 was significantly lower in patients exhibiting lymph node metastasis compared with that in patients lacking lymph node metastasis in human cervical squamous cell carcinoma. Forced expression of HDAC10 in cervical cancer cells significantly inhibited cell motility and invasiveness in vitro and metastasis in vivo. Mechanistically, HDAC10 suppresses expression of matrix metalloproteinase (MMP) 2 and 9 genes, which are known to be critical for cancer cell invasion and metastasis. At the molecular level, HDAC10 binds to MMP2 and -9 promoter regions, reduces the histone acetylation level, and inhibits the binding of RNA polymerase II to these regions. Furthermore, an HDAC10 mutant lacking histone deacetylase activity failed to mimic the functions of full-length protein. These results identify a critical role of HDAC10 in suppression of cervical cancer metastasis, underscoring the importance of developing isoform-specific HDAC inhibitors for treatment of certain cancer types such as cervical squamous cell carcinoma.     

老年宫颈癌50例临床及预后分析

目的：探讨老年宫颈癌的临床病理特征及预后影响因素。方法：对50例老年宫颈癌患者的临床特点及生存情况进行回顾性分析。结果：鳞状细胞癌45例（90．0％）,腺癌4例（8．0％）,透明细胞癌1例（2．0％）;≤Ⅱa期占16．0％,≥Ⅱb期占84．0％;主要临床症状为绝经后不规则阴道流血,全组1、3、5年生存率分别为82．0％,66．0％,54．0％。多因素分析结果显示病理类型、临床分期及淋巴结转移情况是老年宫颈癌患者预后的独立影响因素（均P〈0．05）。结论：KPS评分≥70分、鳞癌、临床分期为Ⅰ期、Ⅱ期,无淋巴结转移的患者预后较好,病理类型、临床分期及淋巴结转移情况是老年宫颈癌患者预后的独立影响因素。     

PRIMARY SQUAMOUS CELL CARCINOMA OF THE FALSE VOCAL CORD

Primary squamous cell carcinoma of the false vocal cord may arise on the surface mucosa or may arise beneath it and continue to grow deeply, presenting only a smooth tumescence of the area. These lesions may not cause hoarseness until late in the course of development. Diagnosis of submucosal primary lesions may present difficulty.Widefield laryngectomy is recommended for small primary lesions of the surface mucosa of the false vocal cord. Such lesions do not show edema of the tissues or deep ulceration and do not cause limitation of motion of the false or true vocal cords.For advanced lesions of the false vocal cord which arise on the surface mucosa and cause edema, ulceration and limitation of motion without enlargement of cervical nodes, widefield laryngectomy and elective block dissection of the neck should be done at the primary operation. Patients with such a primary lesion and metastasis to cervical lymph nodes, which are resectable, should be treated in a like manner.Patients with submucosal primary squamous cell carcinoma of the false vocal cord should be treated with widefield laryngectomy and block dissection of the neck, whether or not palpable resectable lymph nodes are noted in the neck. Apparently these lesions metastasize early and widely.     

Bcl-2和COX-2在宫颈鳞癌中的表达及其临床意义

目的：观察Bcl-2和COX-2在正常宫颈和宫颈鳞癌中的表达情况,并探讨其与宫颈鳞癌发生发展的关系。方法：应用免疫组织化学s-P法检测40例宫颈鳞癌组织、10例正常宫颈组织中Bcl-2和COX-2的表达情况。结果：（1）Bcl-2在正常宫颈组织和宫颈鳞癌组织中的阳性表达率分别为30．0％、72．5％（P〈0．05）,而COX-2在正常宫颈组织和宫颈鳞癌组织中的阳性表达率分别为0．0％、60．0％（P〈0．05）。（2）在宫颈鳞癌中,Bcl-2的表达与宫颈鳞癌的病理分级、临床分期以及淋巴结转移无关（P〉0．05）,而COX-2的表达与宫颈鳞癌的病理分级及淋巴转移有关（P〈0．05）,与临床分期无关（P〉0．05）。（3）Spearman等级相关性分析显示宫颈鳞癌组织中Bcl-2和COX-2的表达呈正相关（r=0．517,P〈0．01）。结论：Bcl-2和COX-2在宫颈鳞癌中的表达升高并呈显著正相关,且COX-2的表达与宫颈鳞癌的淋巴转移有关,二者在宫颈癌的发生发展中可能起重要作用,有可能作为评估宫颈鳞癌淋巴结转移的参考指标。     

A rat model to study the role of STn antigen in colon cancer

Expression of the mucin-associated sialyl-Tn (STn) antigen has been associated with a decreased survival in patients with colorectal, gastric, and ovarian cancer. To better understand the role of STn antigen in tumor biology, we developed STn(+) (called LP) and STn(-) (called LN) clonal cell lines from a parental metastatic rat colon carcinoma cell line (LMCR). Both derivative cell lines exhibited identical proliferation rates in vitro. LP cells strongly expressed STn antigen both in vitro and in vivo, and were poorly tumorigenic when given to syngeneic rats. LN cells did not express STn antigen in vitro, but as in vivo tumors these cells rapidly acquired STn expression, readily formed tumors, and were highly lethal. When rats were given an otherwise lethal inoculum of i.p. LN cells, pre-immunization with synthetic STn antigen conjugated to keyhole limpet hemocyanin (STn-KLH) resulted in a 60% survival rate. When LN cells were injected subcutaneously in the presence of STn-KLH-sensitized lymphocytes, tumor growth was decreased. Distribution of STn antigen in normal organs of host rats is quite similar to that of humans. This model mimics human disease and should facilitate studies of mucin-associated antigens in tumor biology and the development of immunotherapeutic agents based on mucin-related antigens.     

KITENIN increases invasion and migration of mouse squamous cancer cells and promotes pulmonary metastasis in a mouse squamous tumor model

KAI1 C-terminal interacting tetraspanin (KITENIN) is reported to promote metastasis in mouse colon cancer models. We investigated the role of KITENIN on the progression of squamous cell carcinoma (SCC). In a preliminary clinical study using resected tissues from head and neck SCC patients, KITENIN was highly expressed in tumors and metastatic lymph nodes, while KAI1 was more increased in adjacent mucosa than in tumor. KITENIN-transfected mouse squamous cancer (SCC VII/KITENIN) cells showed significantly higher invasion, migration, and proliferation than empty vector-transfected cells. In syngeneic mouse squamous tumor models, more increased tumor volume and enhanced lung metastasis were found in SCC VII/KITENIN cells-injected mice. Thus, KITENIN increases invasion and migration of squamous cancer cells and thereby promotes distant metastasis in mouse squamous tumor models.     

Development of an experimental model for spontaneous lymph node metastasis of human esophageal carcinoma in nude mice--histopathological analysis]

Three human squamous cell carcinoma cell lines (HPL-EsC-1-K, -S, and -M) originated from a male patient with esophageal carcinoma were established and were studied on their tumorigenic and metastatic properties in nude mice. All cell lines grew in the hind foot pads following subcutaneous inoculation and produced popliteal lymph node metastasis dose (2-8 x 10(6)/mouse)-dependently. Based on the histopathological findings on serial sections of the lymph nodes, the stages of lymph node invasion by cancer cells were classified into 4 stages (St. 0-III). The time course of lymph node metastasis of EsC-K cells were examined. Advanced stage of metastasis increased according to the time elapsed after tumor cell inoculation. Incidence of metastasis of EsC-K cells were not affected by host factors such as sex differences, anti-asialo GM1 antibody treatment on the hosts. Today, there are few experimental models for studies on spontaneous lymph node metastasis of human carcinomas. This experimental model provides a useful research tool for studies on the biology and therapy for lymph node metastasis of esophageal cancer.     

Expression of proliferation marker Ki67 correlates to occurrence of metastasis and prognosis, histological subtypes and HPV DNA detection in penile carcinomas

Clinical outcome of penile squamous cell carcinoma (PSCC) largely depends on the presence of lymph node metastasis. In search of a valuable marker predicting the risk for metastasis, the expression of Ki67 was investigated immunohistochemically in primary tumors and compared to presence of inguinal lymph node metastasis. As human papilloma virus (HPV) is thought to affect Ki67 expression, we evaluated whether occurrence of HPV DNA correlates to Ki67 score or metastatic potential. Samples originated from patients subjected to resection of invasive SCC of penis. Immunohistochemistry was done on paraffin-embedded sections using a monoclonal antibody against Ki67. After DNA isolation from paraffin embedded tissue the presence of HPV 6/11, HPV 16 and HPV 18 DNA was analyzed by PCR. Statistical analysis was done using two tail unpaired t test and Chi-square test. Four of 28 patients showed a weak Ki67 expression, without displaying lymph node metastasis. Among 17 patients showing an intermediate Ki67 index, eight exhibited metastases while in all seven patients with a strong expression of Ki67 lymph node metastases were found. The median Ki67 expression in metastastic lesions was significantly different (50.3%) from tumors without lymph node metastasis (31.8%) (p=0.024). Furthermore, a correlation between presence of HPV DNA and strong Ki67 expression was determined (p=0.009). Since our study demonstrated a strong Ki67 labeling index significantly associated to positive lymph nodes, we suggest Ki67 expression as a prognostic marker for lymph node metastasis in penile squamous carcinoma.     

舌鳞癌层粘连蛋白的表达及意义

采用免疫组织化学技术,研究了正常、异常增生组织、舌癌及舌癌淋巴结转移灶中层粘连蛋白的表达、分布及意义。结果发现：正常舌粘膜、轻度异常增生组织的基底膜处层粘连蛋白呈连续线状分布,中度和重度异常增生组织基底膜层粘连蛋白的分布有局部断裂。舌癌中层粘连蛋白的分布呈多种形式,分化好的舌癌层粘连蛋白呈线状,但明显不连续,分化差的舌癌中的层粘连蛋白常呈碎片状,而舌癌的淋巴结转移灶中层粘连蛋白的分布与原发灶相似。统计分析表明,层粘连蛋白的表达与舌癌的分化程度相关,而与舌癌的转移无关。观察结果表明：舌癌分化程度愈低,层粘连蛋白的缺损愈严重,层粘连蛋白的表达、分布特点可以作为判断癌恶性程度的指标,但不能判断舌癌的转移     

Applicability of Preoperative Nuclear Morphometry to Evaluating Risk for Cervical Lymph Node Metastasis in Oral Squamous Cell Carcinoma

Background

We previously reported the utility of preoperative nuclear morphometry for evaluating risk for cervical lymph node metastases in tongue squamous cell carcinoma. The risk for lymph node metastasis in oral squamous cell carcinoma, however, is known to differ depending on the anatomical site of the primary tumor, such as the tongue, gingiva, mouth floor, and buccal mucosa. In this study, we evaluated the applicability of this morphometric technique to evaluating the risk for cervical lymph node metastasis in oral squamous cell carcinoma.

Methods

A digital image system was used to measure the mean nuclear area, mean nuclear perimeter, nuclear circular rate, ratio of nuclear length to width (aspect ratio), and nuclear area coefficient of variation (NACV). Relationships between these parameters and nodal status were evaluated by t-test and logistic regression analysis.

Results

Eighty-eight cases of squamous cell carcinoma (52 of the tongue, 25 of the gingiva, 4 of the buccal mucosa, and 7 of the mouth floor) were included: 46 with positive node classification and 42 with negative node classification. Nuclear area and perimeter were significantly larger in node-positive cases than in node-negative cases; however, there were no significant differences in circular rate, aspect ratio, or NACV. We derived two risk models based on the results of multivariate analysis: Model 1, which identified age and mean nuclear area and Model 2, which identified age and mean nuclear perimeter. It should be noted that primary tumor site was not associated the pN-positive status. There were no significant differences in pathological nodal status by aspect ratio, NACV, or primary tumor site.

Conclusion

Our method of preoperative nuclear morphometry may contribute valuable information to evaluations of the risk for lymph node metastasis in oral squamous cell carcinoma.     

P16、P27蛋白在食管鳞状细胞癌中的表达及意义

目的为探讨P16、P27蛋白在食管鳞状上皮、增生上皮和癌变上皮中表达状况及其与鳞状细胞癌发生、进展和转移的相关性.方法采用SP免疫组织化学方法,检测72例(其中活检标本13例)食管癌组织中P16、P27蛋白的表达情况.结果 P16、P27在食管癌和增生性上皮均有阳性表达,但两者相比均无统计学意义(P>0.05),在正常上皮组均无阳性表达;P16、P27在高分化鳞癌组阳性表达率均显著高于低分化鳞癌组(P<0.05);P16、P27在伴有淋巴结转移的食管癌组与不伴有淋巴结转移组比较差异显著(P<0.05).P16在原发食管癌组阳性表达率和淋巴结转移癌中比较具有显著性差异(P<0.05).结论 P16、P27基因蛋白在食管癌组织中的表达与病理分化程度有关;P16的表达与转移癌的形成有关;P16、P27表达与患者性别、年龄、肿瘤发生部位、浸润深度无明显相关性.