Expression of a Novel β1 Integrin in the Dysplastic Progression of the Cervical Epithelium

Epithelial cell interactions with matrices and basal membranes are central for tissue organization, and integrins are a family of adhesion molecules that play a major role in these interactions. We have analyzed the expression of a novel integrin α chain, α_10.1.2, in the squamous epithelium of the portio vaginalis uteri from patients with cervical intraepithelial neoplasia (CIN) and from control samples without apparent cervical abnormalities. The localization of β₁ and α₆ chains was also investigated, together with the distribution of intraepithelial HLA-DR and CD1c-positive Langerhans cells. In the normal cervical epithelium, all of the integrin chains were detected in the basal cell layer, albeit with a different localization in the apical, lateral, and basal cell surfaces. Langerhans cells were evenly distributed in the deep of the squamous epithelium. Expression of α_10.1.2 was reduced or absent in all of the CIN 1 specimens in which the other integrin chains were either normal or slightly reduced. CIN 2 was characterized by overexpression of integrins, namely of β₁ and of α_10.1.2 chains which were consistently detected also in suprabasal cell layers. None of the integrin chains was found in CIN3 samples in which Langerhans cells were also absent. Thus, modulation and redistribution of integrins occur in the progression of cervical dysplasia, and lack of integrin expression characterizes high-grade lesions. As in other dysplasias and cancers of squamous epithelia, the localization of α_10.1.2 chains provides reliable diagnostic and possibly prognostic criteria.     

E-cadherin and alpha-catenin expression during tumor progression of cervical carcinoma

OBJECTIVE: The objective of this study was to determine whether a relationship between the E-caherin molecule and the E-cadherin-associated cytoplasmic molecule, alpha-catenin, could provide additional information in the neoplastic progression of cervical cancer. METHODS: In this study we investigated by immunohistochemistry E-cadherin (E-cad) and alpha-catenin (alpha-cat) expression in 57 cervical biopsy samples representative of normal exocervical epithelium, viral (human papillomavirus (HPV) infection) and dysplastic lesions (low- and high-grade lesions), and invasive squamous cell carcinoma. The presence of immunostaining and the pattern of distribution of these molecules were correlated with the histological diagnosis (normal cervical epithelium vs HPV, cervical intraepithelial neoplasia (CIN), and invasive carcinoma). RESULTS: The correlation between alpha-cat expression and the histological diagnosis was statistically significant (chi2 test, P < 0.0077); moreover, E-cad and alpha-cat distributions were significantly correlated with the histological classification (P < 0.0001 and P < 0.0043, respectively). CONCLUSION: These results suggest that not only E-cad but also alpha-cat are associated with the loss of differentiation in CIN and squamous cell carcinomas; thus the coexpression of E-cad and alpha-cat may represent a discriminant of malignant potential and could provide an additional criterion to define high-grade CIN.     

Integrin alpha-6 in cervix neck squamous cell carcinoma HPV 16/18 positive and negative]

In the present paper we study expression of alpha-6 integrin in normal paraepidermal epithelium, in CIN and in cervical invasive carcinoma HPV 16/18 positive and negative. The results suggest that alpha-6 integrin might be a marker for the evaluation of malignancy of squamous cell carcinoma. There is no correlation between the expression of alpha-6 integrin and the presence of HPV.     

Laminin-5 gamma 2 chain expression in cervical intraepithelial neoplasia and invasive cervical carcinoma

BACKGROUND. To analyze the expression of laminin-5 gamma 2 chain, a protein which plays a major role in keratinocyte migration, in cervical intraepithelial neoplasia (CIN), and invasive cervical carcinoma associated with high-risk oncogenic human papillomaviruses (HR-HPVs). MATERIAL AND METHODS. The expression of laminin-5 gamma 2 chain protein has been analyzed by immunohistochemistry in 17 cases of low-grade squamous intraepithelial lesions (LSIL-CIN1), 35 high-grade squamous intraepithelial lesions (HSIL-CIN2/3), 18 microinvasive or invasive carcinomas, and three metastatic lymph nodes. All these lesions have been proved to contain HR-HPVs and were also positive for p16 protein which classically is overexpressed at all stages of cervical neoplasia and dysplasia linked with HR-HPVs. 20 cases of normal cervix served as controls. RESULTS. The expression of laminin-5 gamma 2 chain protein was observed in 100% of invasive or microinvasive carcinoma and in their related lymph node metastasis with an immunoreactivity located preferentially at the invasive front of the lesions. All the HSILs (100%) associated with invasive carcinoma were also positive. In contrast, in HSILs without associated invasive component, the expression of the protein has been found in only 34% of cases. In positive HSILs, laminin-5 gamma 2 protein was expressed in basal layers. In LSILs and normal epithelium, no expression of the protein was noted. CONCLUSIONS. We conclude the following: (i) the expression of laminin-5 gamma 2 is a late event in cervical carcinogenesis increasing with the grade of dysplastic lesions; (ii) laminin-5 gamma 2 expression facilitates the identification of invasive and microinvasive lesions which could be difficult to diagnose on the basis of routine stains; (iii) laminin-5 gamma 2 expression in HSILs could potentially identify those lesions with a more increased risk of tumor progression.     

α1β3、ICAM-1、E-cadherin、P-选择素对宫颈癌局部浸润作用的研究

目的探讨细胞黏附分子在宫颈癌局部浸润中的作用。方法应用组织芯片技术和免疫组织化学方法对比检测子宫颈鳞状细胞原位癌30例、微小浸润癌34例、浸润癌36例中α1β3、ICAM-1、E-cadherin和P-选择素的表达。结果α1β3在原位癌中的表达率明显低于微小浸润癌和浸润癌(P<0.05),微小浸润癌与浸润癌的表达率无显著差别(P>0.05);P-选择素的表达率在原位癌、微小浸润癌和浸润癌中无显著差别(P>0.05);ICAM-1、E-cadherin在原位癌中的阳性表达率均显著高于微小浸润癌(P<0.05),微小浸润癌中的阳性表达率均显著高于浸润癌(P<0.05)。ICAM-1、E-cadherin在微小浸润癌、浸润癌中的表达水平密切相关(P<0.01),ICAM-1、E-cadherin的阳性表达与子宫颈癌的浸润呈显著负相关(r=-0.8726,P<0.01和r=-0.7968,P<0.01)。结论α1β3、ICAM-1、E-cadherin与宫颈癌的局部浸润有关,其中ICAM-1、E-cadherin在宫颈癌局部浸润过程中可能起着重要作用。     

Expression of matrix metalloproteinase-2 in preinvasive and invasive carcinoma of the uterine cervix

PURPOSE: To study the immunohistochemical expression of matrix metalloproteinase-2 (MMP-2) in preinvasive and invasive carcinoma of the uterine cervix so as to demonstrate whether the expression of MMP-2 is an early or late event in the process of dedifferentiation and cancer progression. METHODS: A total number of 50 samples of cervical tissue were studied for MMP-2 immunoreactivity. The cases were selected to include ten normal cases used as a control group, 20 CIN cases and 20 cervical carcinoma cases. The CIN group was subdivided into CIN1 (n = 7), CIN2 (n = 6) and CIN3 (n = 7), while the carcinoma group was represented by squamous cell carcinoma (n = 16) and adenocarcinoma (n = 4). RESULTS: MMP-2 expression was totally absent in control cervices and low-grade squamous intraepithelial lesions, while high-grade squamous intraepithelial neoplasia and invasive carcinoma showed up-regulation of MMP-2 expression with no significant difference concerning the type of carcinoma. This overexpression of MMP-2 points to the possibility that it is an early marker of tumor progression in cervical carcinoma. CONCLUSIONS: MMP-2 has a key role in extracellular matrix degradation and invasion in carcinoma of the uterine cervix. Its expression in high-grade squamous intraepithelial lesions may denote a potential risk for invasion and metastasis.     

Expression of glucose transporter-1 in cervical cancer and its precursors

OBJECTIVE: Increased glucose uptake and utilization is a known phenomenon exhibited by malignant cells. Overexpression of the glucose transporter protein family is thought to be the principal mechanism by which these cells achieve up-regulation. Our purpose is to determine glucose transporter-1 (GLUT 1) expression in squamous carcinoma of the cervix and precursor lesions. METHODS: Archival histologic sections were obtained from 31 cases of invasive squamous cell carcinoma (SCC) of the uterine cervix, 15 cases of high-grade cervical intraepithelial neoplasia, 5 cases of low-grade, and 9 normal cervices. Immunohistochemistry for GLUT 1 protein was performed using polyclonal GLUT 1 antibody (Dako, Carpinteria, CA) and the labeled streptavidin-biotin procedure. RESULTS: Compared to the internal control, the pattern of staining varied from weak (1+) to strong (3+) reactions. In normal cervix, 1+ GLUT 1 staining was seen in the basal cells of the squamous epithelium. All 31 (100%) cases of SCC were positive for GLUT 1. Positive reactions seemed more intense in tumor cells that were farther away from the stromal blood supply. There was a correlation between intensity of reaction for GLUT 1 and histologic grade of tumor (P = 0.0027) and with progression from normal or dysplastic lesions to invasive cancer (P = 0.0001). Intensity was a predictor of the presence of poorly differentiated tumor type. Low-grade CIN staining was seen in less than one-third of the epithelium, while in high-grade lesions the reaction was present in over one-half of the epithelium. CONCLUSIONS: GLUT 1 is overexpressed in cervical carcinoma. The process appears to be related to grade of tumor but not to the progression from preneoplastic lesions. The results suggest that GLUT 1 overexpression is a late phenomenon in cellular transformation. Furthermore, the possible relation of expression to tumor blood supply suggests that the malignant cells may have an adaptive environmental ability to compensate for a compromised microenvironment.     

Expression of α4β1 and αvβ3 integrins in the endometrium of women using the T200 copper intrauterine device

Objective: To investigate the expression of α4 and β3 integrin subunit levels in the endometrium of healthy women and copper intrauterine device (IUD) T200 users.Design: Case control study.Setting: An academic teaching hospital and a primary care clinic.Patient(s): Thirteen copper IUD users and 13 normal fertile women.Intervention(s): Timed endometrial biopsies during the mid-secretory phase (days 20 to 24).Main Outcome Measure(s): Histologic dating of endometrium and immunohistochemical staining intensity of α4 and β3, using the semiquantitative immunohistochemical score (HSCORE).Result(s): All endometrial biopsies consistent with menstrual dates were examined for integrin expression (β3 and α4). No difference in α4 integrin expression was found between IUD users and controls in both luminal and glandular epithelium. In fertile controls, αvβ3 staining was present in 100% and 38.4% of glandular and luminal epithelium, respectively. In contrast, only 61.5% of the IUD users had any αvβ3 staining in the glandular epithelium and only 53.9% in the luminal epithelium. The intensity of immunoreactivity between the two groups (mean HSCORE) did not differ significantly.Conclusion(s): Proportionately, significantly fewer women using copper IUD had positive αvβ3 immunoreactivity in the glandular epithelium of mid-secretory endometrium.     

Alterations in the expression of selected MHC antigens in premalignant lesions and squamous carcinomas of the uterine cervix

BACKGROUND: The objective of the study was to determine whether there is a correlation between expression of selected major histocompatibility complex (MHC) antigens and the presence of carcinoma and premalignant lesions of the cervix. We also attempted to determine whether there is a correlation between expression of the selected MHC antigens in each grade of cervical intraepithelial neoplasia (CIN) and invasive carcinoma of the cervix. METHODS: Quantitative comparative analysis of MHC class I and class II expression between specimens of the uterine cervix was carried out. For detecting human leukocyte antigen (HLA) class I molecules, two primary antibodies were used, recognizing either epitopes of all free heavy chain class I molecules or epitopes of the HLA-Bw4 antigen. HLA class II antigens were detected by antibodies reacting with the alpha-chain of the HLA-DR molecule. An indirect immunoperoxidase technique was carried out using paraffin-embedded sections. The following groups of sections were formed: squamous metaplasia, CIN of different grades, and invasive carcinoma. The specimens were taken from women in whom routine histopathologic diagnosis of cervical lesions had been performed. RESULTS: A significant decrease in the mean number of stained cells showing expression of MHC class I antigens (HC 10 and Bw4) was found in the invasive cancer group of specimens. A decrease in Bw4 expression was also found in the CIN III group. The highest expression of the HLA-DR antigens was detected in the metaplastic epithelium compared to the other groups. In the dysplastic epithelium the expression of the HLA-DR antigens was increased in the highest grade of dysplasia (CIN III). Upregulation of HLA-DR expression was detected in invasive cancer specimens. CONCLUSIONS: Alterations in MHC expression are found in premalignant lesions and squamous carcinomas of the uterine cervix. Quantitative assessment with the use of an image analysis system is valuable in helping to discriminate staining patterns of HLA expression. Further studies are indicated to evaluate the prognostic value of the expression of selected MHC antigens.     

阴道镜引导活检诊断宫颈病变的优势与不足

目的:评价阴道镜引导宫颈活检(CDB)诊断宫颈病变的优势与不足。方法:1258例患者行宫颈LEEP术前均行CDB,比较两者病理组织学诊断的符合率,计算CDB对高级别宫颈病变的漏诊率及CDB诊断高级别宫颈病变的阴性预测值。结果:CDB和LEEP病理组织学诊断完全相符占42.5%,CDB诊断病变级别更高占42.1%,更低占15.4%。CDB准确性为84.6%。CDB诊断为低级别CIN经LEEP术后病理证实21.3%存在高级别CIN。CDB诊断为低级别CIN和高级别CIN实际上存在宫颈浸润癌的比例为0.9%及4.4%,两者有显著差异(P<0.05)。CDB诊断高级别CIN的阴性预测值为78.5%;CDB用于诊断宫颈浸润癌漏诊率为2.2%。结论:阴道镜检查和CDB,可以发现绝大部分的宫颈病变患者。单纯点活检不能排除宫颈浸润癌。CDB诊断为高级别宫颈病变时,应考虑有更重病变的可能,LEEP可弥补CDB的不足,最大限度地提高宫颈疾病诊断的准确性。     

Detection of human papillomavirus DNA in human cervical lesions by tissue in situ nucleic acid hybridization

Cervical cancer is one of the most common female cancers in Taiwan. Certain types of human papillomavirus (HPV) are frequently detected in the epithelial precancerous and cancerous lesions of the cervix. By the use of tissue in situ hybridization, we investigated the relationship of various types of HPV (group I, HPV-6 & 11, group II, HPV-16 & 18, group III, HPV-31, 33 & 35) with cervical condyloma, carcinoma as well as precancerous lesions. Group I HPV DNAs were mainly found in cervical condylomatous lesions (2/2) of the cervix and cervical intraepithelial neoplasia I (CIN I) (2/4), but were only occasionally found in CIN II (1/4), CIN III (1/9) or non-keratinized squamous cell carcinoma (1/15). HPV DNAs of groups II and III were mainly detected in lesions of CIN III (5/9) and invasive squamous cell carcinoma (large cell, keratinized type: 4/7; large cell, non-keratinized type: 11/15). HPV DNA sequences were invariably detectable only in the cell nuclei of condyloma or dysplastic epithelium or invasive carcinoma. However, they could not only be detected in the upper layer dysplastic cells and koilocytes but also in the well and poorly differentiated cervical cancer cells. The distribution of HPV DNA positive cells in the carcinomas fell into four different patterns: (1) upper zone and non-invasive regions of the carcinoma (11/22, 50%), (2) basal zone and invasive regions (2/22, 9%), (3) randomly scattered (7/22, 32%), and (4) extensively distributed over the whole tumor lesions (2/22, 9%). Thus, our results are consistent with a strong correlation between the presence of HPV-16, 18, 31, 33 and 35 and malignant conversion of cervical epithelial cells.     

MMP-1 and MMP-2 in the cervix uteri in different steps of malignant transformation--an immunohistochemical study.

OBJECTIVES: Enzymatic degradation of the extracellular matrix (ECM) represents a key element in the multistage process of tumor invasion and metastasis. This process requires extensive degradation of ECM components such as basement membrane collagen (type IV) and interstitial collagen (type I, II, III). Matrix metalloproteinase-2 (MMP-2) specifically cleaves collagen type IV, the major collagen of the basement membrane. MMP-1 digests interstitial collagen type I and III, the main collagen types of the stromal extracellular matrix. We investigated protein levels of MMP-1 and MMP-2 in different stages of malignant transformation. METHODS: Using the APAAP method we analyzed 10 normal cervical tissues, 11 cervical intraepithelial neoplasia 1 (CIN 1), 8 CIN 2 and 10 CIN 3 lesions, and 15 invasive squamous cell carcinomas. These data were compared with the HPV DNA status tested by hybrid capture II. RESULTS: Only a few isolated epithelial cells stained positively for MMP-1 and MMP-2 in normal cervical tissue and CIN 1 lesions. The CIN 2 and CIN 3 group displayed a heterogeneous distribution of MMP expression. 3 CIN 2 and 8 CIN 3 lesions showed strong MMP-2 and weak MMP-1 expression in the dysplastic epithelial cells. 5 CIN 2 and 2 CIN 3 lesions stained negatively. Invasive carcinomas showed a coexpression for MMP-1 and MMP-2 in malignant epithelial cells and peritumoral stroma cells. All MMP-2-positive cases tested positive for the HPV high-risk group. CONCLUSIONS: The expression of MMP-2 protein in preinvasive lesions of the cervix uteri and a consecutive coexpression of MMP-1 and MMP-2 in invasive cancer suggest a gradually increasing invasive potential. MMP-2 expression, when focally observed in high-grade squamous intraepithelial lesions of the cervix, may indicate tumor areas with an increased risk for invasive growth.     

CD1a和E-cadherin在宫颈上皮内瘤变中的表达

目的:探讨CD1a和E-cadherin与宫颈癌发生发展的关系及作为早期癌变生物学指标的可能性。方法:采用免疫组化SP法检测CD1a和E-cadherin在同期56例宫颈上皮内瘤变、56例宫颈鳞癌和15例正常宫颈组织中的分布及表达。结果:(1)CD1a+朗格汉斯细胞在正常宫颈、CIN和宫颈癌各组中数量逐渐减少,两两比较有显著差异(P0.01);CIN中该细胞数量随病变严重程度减少,CINⅠ和CINⅡ、CINⅠ和CINⅢ组之间两两比较有显著差异(P0.05),而CINⅡ与CINⅢ组间无显著差异(P0.05);(2)E-cadher-in在正常宫颈、CIN和宫颈癌各组中的阳性表达率及强度逐渐下降,两两比较有显著差异(P0.05);在CIN中阳性表达率及表达强度随病变严重程度呈下降趋势,CINⅠ和CINⅡ、CINⅠ和CINⅢ之间两两比较有显著差异(P0.05),而CINⅡ和CINⅢ之间无显著差异(P0.05);(3)宫颈组织中CD1a+朗格汉斯细胞的细胞数与E-cadherin的阳性表达率及强度呈正相关(r=0.912,P0.05)。结论:CD1a+朗格汉斯细胞与E-cadherin可能在宫颈癌的发生、发展过程中起重要作用。     

Immunohistochemical localization of filaggrin in benign,premalignant and malignant cervical tissue

Epithelial distribution of filaggrin, a histidine-rich protein related to squamous terminal differentiation, was investigated in 87 cervical biopsies using an avidin-biotin-peroxidase technique with a monoclonal anti-human filaggrin antibody (AKH1). Normal squamous cervical epithelium exhibited a positive homogeneous immunoperoxidase stain in the upper parabasal, intermediate and superficial cell layers. Similar findings were obtained in cervical condylomas, although full-thickness staining was observed in 35.7% of the cases (P<0.001). Filaggrin expression in CIN was inversely related to the severity of the lesion (P<0.001). An irregular staining pattern was present in most high-grade CIN. Filaggrin expression was closely connected to the degree of tumour differentiation (P<0.05) in squamous cell carcinomas of the cervix. Abnormal filaggrin stainings identified a premalignant/malignant cervical squamous lesion with a positive predictive value of 92.3%. Non-squamous epithelia showed lack of filaggrin expression. Filaggrin may therefore be considered a marker of squamous differentiation in both the normal and pathological human uterine cervix.     

细胞凋亡和P53在宫颈癌及宫颈上皮内瘤变中的表达

目的：探讨细胞凋亡和P53在宫颈癌及宫颈上皮内瘤变中的作用。方法：用DNA缺口末端标记技术(TUNEL)和免疫组化技术原位观察15例正常宫颈、22例CIN*"I/Ⅱ、16例CINⅢ和52例浸润性宫颈鳞癌(ISCC)组织中细胞凋亡标记指数(A-LI)、PCNA(增殖细胞核抗原)标记指数(PCNA-LI)和P53表达阳性率及P53标记指数(P53-LI)。结果：在正常宫颈上皮未检测到凋亡细胞,随着CIN病变的进展,A-LI增高(P＜005),在ISCC,A-LI明显低于CIN(P＜001);PCNA-LI随宫颈癌各级病变的进展逐渐增高(Ｐ＜005);在正常宫颈和CINI/Ⅱ均未检测到P53表达,在CINⅢ,P53-LI明显低于ISCC(P＜001)。结论：细胞凋亡在宫颈癌发生发展过程中发挥重要作用,P53蛋白的积聚可能对细胞凋亡起抑制作用。     

E-cadherin expression during progression of squamous intraepithelial lesions in the uterine cervix

Alterations of E-cadherin expression in cervical intraepithelial neoplasias and invasive carcinomas of the uterine cervix have been described by some authors but their clinical significance has not yet been clarified. Archival specimens of 27 normal cervical epithelia, 15 atypical cells of undetermined origin (ASCUS), 53 low-grade squamous intraepithelial lesions (LSIL), 19 high-grade squamous intraepithelial lesions (HSIL) and six invasive squamous carcinomas were evaluated for E-cadherin expression. The cytological material was processed using liquid based cytology (ThinPrep technique) and immunostained for E-cadherin. All HPV infections (koilocytes) showed strong cell membranous E-cadherin expression. In HSIL a strong decrease in E-cadherin expression and heterogeneous distribution was noticed. In the relatively small number of squamous cell carcinomas of the cervix studied, a significant decrease or loss in E-cadherin expression, predominantly cytoplasmic, was noted. We concluded that decreased E-cadherin expression appears to be a useful parameter of malignant potential of cervical lesions. E-cadherin immunoexpression could provide an additional criterion in correlation with cyto- and histomorphology and colposcopy to define high grade CIN lesions.     

不同级别卵巢浆液性癌中E-cadherin和p120蛋白的表达及意义

目的:探讨上皮性钙黏附蛋白(E-cadherin)和连环蛋白p120在不同组织学级别卵巢浆液性癌中的表达及意义。方法:用免疫组化EnVision二步法,分别检测31例卵巢低级别浆液性癌和70例卵巢高级别浆液性癌中E-cadherin和p120蛋白的表达水平,分析其与卵巢浆液性癌临床病理资料的相关性。结果:E-cadherin在低级别浆液性癌和高级别浆液性癌中的阳性率分别为71.0%和61.4%(P=0.356);p120的阳性率分别为83.9%和60.0%(P=0.018)。高级别浆液性癌中,E-cadherin和p120蛋白表达率随着FIGO分期(P=0.015和P=0.011)、淋巴结转移(P=0.001和P=0.001)及大网膜受累(P=0.028和P=0.018)显著降低。E-cadherin蛋白表达率随年龄增高显著降低(P=0.011),而p120蛋白表达率与年龄无显著相关性(P=0.226)。低级别浆液性癌中,仅p120蛋白表达率与淋巴结转移(P=0.042)、大网膜受累(P=0.048)及术前CA125水平(P=0.042)具有显著相关性。结论:卵巢高级别浆液性癌中p120蛋白的表达缺失率显著高于卵巢低级别浆液性癌,同时仅在高级别浆液性癌中E-cadherin和p120的低表达均与FIGO分期和肿瘤转移潜能相关,提示E-cadherin和p120蛋白可能在不同级别卵巢浆液性癌的发生发展中具有不同的作用。