Subcellular distribution of androgen receptors in human normal, benign hyperplastic, and malignant prostatic tissues: characterization of nuclear salt-resistant receptors

Two populations of nuclear androgen receptors have been characterized in human prostatic tissue, and the levels and proportions of each were found to differ in normal prostates, benign hyperplastic prostates (BPH), and malignant prostates. A significant percentage (35 to 50%) of total nuclear androgen receptors was associated with the salt-resistant nuclear matrix fraction. The remainder were easily extracted from nuclei by 0.6 M KCl. Optimal conditions for measuring receptors in both compartments involved the use of an inhibitor of proteolysis (phenylmethylsulfonyl fluoride) and the omission of dithiothreitol from buffers. In the presence of dithiothreitol, most of the nuclear salt-resistant receptors were rendered salt extractable. Cytosol androgen receptor levels were not significantly different in normal, BPH, or malignant prostatic tissues. In contrast, the levels and distribution of nuclear salt-extractable and salt-resistant androgen receptors exhibited characteristic patterns. Compared to normal prostatic tissue, nuclear salt-extractable receptors were significantly elevated in both BPH and cancer, whereas nuclear salt-resistant receptors were elevated in BPH but not in cancer. The ratio of salt-extractable to salt-resistant receptors was approximately 1:1 in both normal and BPH tissues and 2:1 in cancer. In addition, a microassay has been developed for the measurement of androgen receptors in the three subcellular compartments of needle biopsy specimens of prostatic cancer. Studies are in progress to determine whether the measurement of both nuclear salt-extractable and salt-resistant receptors may improve the usefulness of receptor levels to predict the hormonal responsiveness of prostatic cancer.     

乙酰肝素酶在胃癌、癌旁组织及正常胃组织中的表达及其意义

目的:探讨胃癌、癌旁组织及正常胃组织内乙酰肝素酶mRNA的表达情况及其与胃癌临床病理特征的关系。方法:应用半定量逆转录聚合酶链反应(RT-PCR)以β-actin基因碱基序列为内参照,检测8例正常胃组织、56例胃癌组织及相应癌旁组织内乙酰肝素酶mRNA的表达。结果:正常胃组织未检测到乙酰肝素酶mRNA的表达;56例胃癌患者中表达阳性41例(73.2%),癌旁组织仅有15例(26.8%)表达阳性,其表达值30.44%±10.46%显著低于胃癌组织的59.37%±15.11%,(P<0.001)。胃癌组织中乙酰肝素酶mRNA表达阳性率与肿瘤直径大于5cm、浸润超过肌层,有静脉侵犯、有淋巴结转移、有远处转移以及临床分期为Ⅲ-Ⅳ期者有关,而且表达值也与上述临床病理特征显著相关(P<0.05或P<0.01)。结论:胃癌组织中乙酰肝素酶高表达与肿瘤的生长、浸润和转移相关。     

Expression of LeY and extended LeY blood group-related antigens in human malignant, premalignant, and nonmalignant colonic tissues

The LeY determinant, a difucosylated type 2 blood group-related antigen, is a positional isomer of the Leb blood group antigen and a fucosylated derivative of the LeX antigen. The LeX antigen behaves like an oncodevelopmental tumor-associated antigen in human colon cancer, and extended polyfucosyl LeX antigens are more specific for colon cancer tissues than are simple, monofucosyl LeX antigens. The present investigation compared the expression of simple and extended LeY antigens in a variety of malignant and nonmalignant human colonic tissues to gain insight into the normal distribution and cancer-associated expression of these antigens. Monoclonal antibody AH-6, which recognizes the LeY epitope irrespective of its carrier carbohydrate chain, stained the majority of specimens regardless of malignant potential or location within the colon. In contrast, CC-1 and CC-2 monoclonal antibodies, which recognize extended LeY structures, and KH-1, which is specific to trifucosyl LeY, preferentially stained malignant colonic tissues and rarely stained normal colonic mucosae. Mucosa immediately adjacent to cancer usually stained with AH-6 but not with KH-1, CC-1, or CC-2. Extended or trifucosyl LeY antigen expression was limited exclusively to premalignant (adenomatous) polyps and was invariably absent from nonpremalignant (hyperplastic) polyps. Moreover, among adenomatous polyps, extended LeY antigen expression tended to correlate with three parameters of malignant potential: larger polyp size; villous histology, and severe dysplasia. AH-6 failed to distinguish between hyperplastic and adenomatous polyps. In second-trimester fetal colonic mucosa, AH-6 bound to both proximal and distal segments whereas KH-1, CC-1, and CC-2 bound only to proximal segments. We conclude that in human colon, the LeY hapten is an oncodevelopmental cancer-associated antigen and extended LeY antigens are highly specific markers for malignancy and premalignancy.     

Expression of cell cycle control proteins in normal epithelium, premalignant and malignant lesions of oral cavity

In this study, we examined the expression of cyclins, cyclin dependent kinase (CDKs) and CDK inhibitors by immunohistochemical analysis in 20 normal mucosa, 42 epithelial dysplasia (ED), and 117 oral squamous cell carcinoma. Neither Cyclin D1 nor CDK2 were detectable in normal tissue and ED. Their presence, however, was detectable in squamous cell carcinoma (SCCs) (Cyclin D1, 35.9%; CDK2, 66.7%). Cyclin E was detectable in 57.1% of severe ED and 62.8% of SCCs. For the CDK inhibitors, these proteins were detectable in all normal mucosa and most of the mild and moderate ED. For severe ED, expression of these proteins was not observed in some cases (p12(DOC-1), 14.3%; p16(INK4A), 28.6%; p27(KIP1), 7.1%). For SCCs, the expression of p12(DOC-1) was lost in 71.8%, p16(INK4A) in 69.2% and p27(KIP1) in 35.9%. These results suggest that elevated expression of cyclin D1, cyclin E, CDK2 and loss of p12(DOC-1), p16(INK4A) and p27(KIP1) may contribute to the multistep nature of oral carcinogenesis.     

Immunohistochemical demonstration of epidermal growth-factor receptors in normal, benign and malignant thyroid tissues

Human epidermal growth factor receptor (EGF-receptor) has been detected immunohistochemically in normal, benign and malignant human thyroid tissues. With a monoclonal antibody for EGF-receptor and avidin-biotin-peroxidase complex (ABC), the expression of EGF-receptor was evaluated in paraffin-embedded sections. Carcinomas of the thyroid showed a moderate to intense staining for EGF-receptor in most cases. Apical cell surface and cytoplasmic staining was the most common pattern of immunoreactivity. Adenomas showed a variable positivity in the cytoplasm of th tumor cells, and their apical cell surface staining was generally negative to borderline. The follicular cells in Hashimoto's thyroiditis showed a weak to moderate cytoplasmic staining, but those in Graves' disease and normal thyroids showed an essentially negative cytoplasmic staining. Apical cell surface staining was essentially negative or borderline in these benign lesions. There were no significant correlations between EGF-receptor expression and tumor size, degree of invasion or cervical metastases in the thyroid carcinomas. The apical surface expression of EGF-receptor was characteristic to thyroid carcinomas and this feature may be useful in the differentiation of thyroid carcinomas from benign thyroid lesions.     

Expression of p53 in premalignant and malignant squamous epithelium.

Using three antibodies (JG8, CM-1 and 1081) directed to the p53 protein, strong positivity was found in 16/47 (34.0%) of mucosal squamous cell carcinomas of the head and neck and in two squamous carcinoma cell lines (LICR-LON- HN5 and HN6Rr). The presence of the mutant p53 was confirmed in the cell lines as substitutions in exon 7 (codon 238, TGT greater than AGT) and exon 5 (codon 152, CCG greater than CTG) respectively. Positive staining was seen only in the undifferentiated cells and progressively lost as the cells keratinized, both in the tumour specimens and in the cell lines. Similar results were seen in areas of dysplasia, well removed from the site of the primary tumour. Staining of epidermal lesions showed positivity in 2/12 (16.6%) cases of Bowen's disease, 0/12 (0.0%) cases of solar keratosis, 0/10 (0.0%) basal cell carcinomas and in 3/20 (15.0%) squamous cell carcinomas. These results are discussed in relation to the multifocal origin of squamous cell carcinomas, the role of p53 mutations in squamous cell carcinomas from different sites and the significance of the 'basal' distribution of p53 as a normal growth regulator. The possible significance of the distribution of p53 in squamous epithelium as it relates to papilloma virus infection is also considered.     

Vitamin D receptor expression in normal, premalignant, and malignant human lung tissue.

BACKGROUND: There is a strong interest in identifying chemopreventive agents that might help decrease the burden of lung cancer. The active metabolite of vitamin D, 1,25-dihydroxycholecalciferol (calcitriol), has been shown to have antiproliferative effects in several tumor types, mediated by the vitamin D receptor (VDR). This is the first comprehensive survey of VDR expression in a series of human lung tissues, including normal and premalignant central airway biopsies and lung tumors. METHODS: Immunohistochemical expression of nuclear and cytoplasmic VDR was examined in 180 premalignant or malignant bronchial biopsies from bronchoscopy of 78 high-risk individuals at the Roswell Park Cancer Institute and also in 63 tumor samples from 35 lung cancer patients from the University of Chicago Hospitals. Associations between clinicopathologic data and VDR expression were examined. RESULTS: VDR expression was present in many samples. In biopsies, VDR was commonly detected throughout the full epithelial layer. Most histologically normal (60%, 53 of 88) and metaplastic (61%, 39 of 64) samples had moderate to high nuclear intensity; dysplastic samples mostly had low nuclear intensity (10 of 18, 55%). In tumor samples, 62% (38 of 61) were lacking cytoplasmic VDR, with nuclear expression present in 79%(49 of 62). Analysis of all samples revealed a positive linear trend between proportion of samples with greater nuclear than cytoplasmic intensity and increasing histologic grade (P < 0.01). CONCLUSIONS: VDR expression spanned the lung carcinogenesis spectrum. Nuclear expression was similar across various histologies, whereas cytoplasmic expression decreased with increasing histologic grade. These results indicate that there is potential for the use of calcitriol as a chemopreventive agent against the development of lung cancer.     

Estrogen receptors in human gastric, hepatocellular, and gallbladder carcinomas and normal liver tissues

Steroid binding assay using the dextran coated charcoal (DCC) method was applied to human tissues including tumors of the digestive organs, and the results were compared with those of enzymeimmunoassay (EIA) and immunocytochemical assay (ICA) with monoclonal antibody against human estrogen receptor of MCF-7 breast cancer cells. Using the DCC method, estrogen receptor activity was detected in 6 of 26 cases (23.1%) with gastric carcinoma, 3 of 16 hepatocellular carcinoma cases (18.8%), 1 of 3 gallbladder carcinoma cases (33.3%), and both of the 2 cases (100%) with normal liver tissue. However, using EIA, no ER activity was detected in any case. Moreover, ER positive cells were not found by immunohistochemical staining in the gastric carcinoma cases or in normal liver tissue, both of which showed ER activity by the DCC method. These results suggest that the estrogen receptor like material exists in cytosol of the human digestive tumors and normal liver tissue, but that the specificity of the antibodies against estrogen receptor molecules in these tumors may be different from that of the breast tumors.     

Discrimination between normal and malignant human gastric tissues by Fourier transform infrared spectroscopy

The aim of this study was to determine whether malignant and normal human gastric tissues can be distinguished by Fourier transform infrared (FTIR) spectroscopy. Compared with normal tissue, malignant tissues showed significant increases in infrared (IR) absorption in 10 bands lying in a region of 925-1660 cm(-1). Using the 10 IR absorption bands as markers, discriminant analysis was carried out for tissue discrimination. As a result, 22 out of the 23 gastric cancer samples and 9 out of the 12 gastric normal samples were correctly segregated, yielding 88.6% accuracy. The present results suggest that FTIR spectroscopy is a useful tool for screening gastric cancer.     

Differential expression of retinoic acid receptors in normal and malignant esophageal tissues

The chemopreventive and chemotherapeutic activities of retinoids may be attributed to their ability to modulate growth, differentiation and apoptosis of epithelial cells, suppress or reverse epithelial carcinogenesis. Many of these effects of retinoids result from modulation of genes by two distinct classes of retinoid receptors: RARs and RXRs, alterations in their expression may lead to tumorigenesis. To determine whether alterations in expression of retinoid receptors are related to the development of esophageal squamous cell carcinomas (ESCCs), the expression of RARalpha, beta, gamma and RXRalpha was studied in 50 untreated primary esophageal carcinomas and 19 distant normal tissues by immunohistochemistry. RARbeta expression was observed in 18/50 (36%) ESCCs, while 16/19 (84%) of matched histologically normal esophageal tissues displayed RARbeta immunopositivity (p=0.001, 0R=3.405). Significant increase in RARalpha immunopositivity was observed in ESCCs (40/50; 80 %) as compared to normal tissues (9/19 cases; 47%) (p=0.008; 0R=2.77). RARgamma expression was observed in ESCCs (37/50cases; 74%) as compared to normal tissues (16/19; 84%); without significant difference. However, poorly differentiated esophageal cancer showed marked decrease in RARy immunopositivity (p=0.017; OR=6.0). Interestingly, increased expression of RXRalpha was observed in 43/50 (86%) ESCCs in comparison with (10/19; 53%) normal tissues (p=0.003; 0R=3.09). Logistic regression analysis revealed RARgamma-/RXRalpha+ phenotype as most significantly associated with dedifferentiation of the tumor (p=0.014; OR=11.0). The hallmark of the study was the significant increase in expression of RARalpha and RXRalpha proteins and loss of expression of RARbeta protein in ESCCs in comparison with the distant normal epithelia.     

Expression of two antigens defined by monoclonal antibodies in normal, benign and malignant human mammary tissues

We examined the reactivity of two monoclonal antibodies (MAbs), MBrI and MBr8, on sections from normal breast tissues, benign lesions, primary infiltrating carcinomas and distant breast cancer metastases and compared the expression of the recognized antigens (CaMBr1 and CaMBr8 respectively) in these different normal and pathologic conditions. The expression of both antigens was found to significantly decrease in the neoplastic tissues, in comparison to normal breast tissue (34% vs 61% for MBr1 and 52% vs 86% for MBr8; p less than 0.001 for both). Atypical epithelial hyperplasia was found to be reactive on the same level (31%) of the tumors when tested with MBr1, whereas its reactivity was similar to that of normal tissues (78%) when MBr8 was used. Nonatypical epithelial hyperplasia showed the same reactivity with both MAbs (positive staining on about 50% of the cells). 65% of the "in situ" ductal carcinoma cases was MBr1-positive, whereas 50% was MBr8-positive; these values, when compared with the reactivity on normal glands, were significantly lower for MBr8, but not for MBr1. When testing sections from premenopausal women who had biopsies performed on different days during their menstrual cycle a correlation was found between the expression of CaMBr1 in normal breast epithelial cells and the stimulatory phase of the cycle. These findings suggest that both antigens are typical of normal breast and their expression could possibly decrease in pathologic tissues.     

Expression of insulin-like growth factors (IGFs), their receptors and IGF binding protein-3 in normal, benign and malignant smooth muscle tissues.

To assess the role of insulin-like growth factors (IGFs) in growth and transformation of normal (myometrium) and tumorous smooth muscle cell (SMC) tissues, in situ hybridization (ISH) analysis for insulin-like growth factor I and II (IGF-I and IGF-II) mRNAs was combined with detection of IGF peptides, their receptors and IGF binding protein-3 (IGFBP-3). mRNAs for both IGFs were detected in smooth muscle cells in normal, benign and malignant SMC tissues, together with the IGF peptides, both IGF receptors and IGFBP-3. This suggests an autocrine role for both IGFs. Leiomyomas had higher IGF-I peptide levels and higher levels of type I IGF receptors than myometrium, supporting the idea that IGFs play a role in the growth and transformation of these tumours. Low-grade leiomyosarcomas contained more IGF-II mRNAs than myometrium and leiomyoma, fewer type II IGF/mannose 6-phosphate receptors and less IGFBP-3 than myometrium and, in addition, fewer IGF-I mRNAs and type I IGF receptors than leiomyoma. Intermediate- and high-grade leiomyosarcomas had intermediate levels of IGF-II mRNAs and peptide, ranging between those in myometrium and low-grade leiomyosarcomas. Thus, growth and transformation of leiomyosarcomas may be regulated by IGF-II, although more markedly in low-grade than in high-grade leiomyosarcomas. In conclusion, the various categories of SMC tissues are associated with a distinct expression pattern of the IGF system. This suggests that each category of SMC tumours arises as a distinct entity and that there is no progression of transformation in these tissues.     

Expression of Bcl-2 family member Bid in normal and malignant tissues

Bid is the only known Bcl-2 family member that can function as an agonist of proapoptotic Bcl-2-related proteins such as Bax and Bak. Expression of the proapoptotic Bcl-2 family protein Bid was assessed by immunoblotting and immunohistochemical methods in normal murine and human tissues, and in several types of human cancers and tumor cell lines. Bid expression in normal tissues varied widely, with prominent Bid immunostaining occurring in several types of short-lived cells (e.g., germinal center B cells, peripheral blood granulocytes, differentiated keratinocytes) and in apoptosis-sensitive cells (e.g., adult neurons). Analysis of Bid expression by immunostaining of 100 colon, 95 ovarian, and 254 prostate cancers, as well as 59 brain tumors and 50 lymphomas, revealed evidence of altered Bid regulation in some types of cancers. Correlations with clinical outcome data revealed association of higher levels of Bid with longer recurrence-free survival in men with locally advanced (T3 stage) prostate cancer (P=0.04). Immunoblot analysis of Bid protein levels in the NCI's panel of 60 human tumor cell lines revealed a correlation between higher levels of Bid and sensitivity to ribonucleotide reductase (RR)-inhibiting drugs (P<0.0005). Overexpression of Bid in a model tumor cell line by gene transfection resulted in increased sensitivity to apoptosis induction by a RR inhibitor. Taken together, these observations suggest a potential role for Bid in tumor responses to specific chemotherapeutic drugs, and lay a foundation for future investigations of this member of the Bcl-2 family in healthy and diseased tissues.     

Expression of cytokines and receptors in normal,immortalized, and malignant ovarian epithelial cell lines

BACKGROUND: Ovarian carcinoma originates from the epithelial cells on the surface of the ovary. This study evaluates cytokine production by these cells. MATERIALS AND METHODS: Normal human ovary surface epithelial cells (HOSE cells), immortalized HOSE cells and ovarian cancer cells were used for the study of cytokines. RESULTS: Eight of 14 cytokines were increased in > or = 3 ovarian cancer cell lines compared with normal HOSE cells. Three cytokines were increased 5-fold in the immortalized HOSE cell line and in multiple ovarian cancer cell lines. Cytokine receptor expression revealed that 7 of 8 ovarian cancer cell lines had > or = 1 autocrine loop. Anti-EGFR antibody failed to inhibit growth of ovarian cancer cells which expressed multiple cytokine receptors. CONCLUSION: Ovarian cancer cells produce more cytokines than normal HOSE cells. Immortalized HOSE cells display a cytokine profile similar to the cancer cells. Finally, multiple autocrine loops in ovarian cancer may limit the therapeutic usefulness of single cytokine receptor blockade.     

Expression of gastric-associated antigens by human premalignant and malignant colonic epithelial cells

A rabbit antiserum prepared to 2nd-trimester fetal organ extracts and absorbed with adult tissue (anti-STFa) was used to detect antigens common to 2nd-trimester fetal large bowel, normal adult stomach, several colon carcinoma cells (in primary and established cultures) and epithelial cells cultured from benign colonic polyps. The frequency of anti-STFa-positive cells was highest in cultures derived from benign tumors known to be associated with a greater degree of premalignancy. Thus, the percentage of antigen-positive cells increased from 18 and 43% to 70% of cultures from tubular, villotubular and villous adenomas, respectively. Considerable heterogeneity in the distribution of antigen-containing cells was evident within any given area of a positive culture. Absorption experiments, using a spectrum of fetal and normal adult tissue extracts, indicated that the adenoma-carcinoma specificity resides in fetal, but not adult, large bowel and normal adult stomach.     

原位杂交法检测SNCG在乳腺组织中的表达

目的:检测乳腺癌相关基因SNCG在乳腺组织不同病变的阳性表达率,分析其表达规律,探讨其在乳腺癌发生、发展中的作用。方法:用原位杂交方法,以地高辛标记的SNCG反义mRNA作探针,检测167例乳腺不同病变组织中SNCG基因mRNA的表达情况,并进行统计学分析。结果:SNCG的阳性表达率在乳腺增生症为76.0%(38/50),纤维腺瘤为20.0%(3/15),非典型性增生为92.3%(12/13),乳腺癌组织为95.5%(85/89),并且表达强度不同。有淋巴结转移的乳腺癌中强阳性表达率为47.7%(21/44)。而乳腺增生症组中无强阳性表达者。SNCG在不同乳腺病变组织中的表达有统计学意义(P<0.001)。结论:SNCG作为一个新发现的乳腺癌相关基因,在乳腺不同病变中表达情况不同,随病变程度加重,表达率明显增高。有转移者表达强度明显增强。因而SNCG可作为判断乳腺癌恶性潜能的指标。