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C D Albright

Publications and source records attributed to C D Albright.

At least 37 records · Page 2Linked to original sources
PubMed1995

Human endometrial carcinoma cells release factors which inhibit the growth of normal epithelial cells in culture.

Autocrine and paracrine interactions between cells are important homeostatic mediators in normal tissues. Alterations to growth factor signalling pathways are likely to play a role in multistep carcinogenesis. In this study normal human endometrial epithelial cells (NHEC) after 3 days in culture were treated with serum-free medium conditioned for 24 h by log phase or confluent cultures of established RL95-2, HEC1A, or AN3CA endometrial carcinoma (EC) cell lines. By day 4, NHEC treated with either log phase or confluent conditioned medium (CM) showed a significant decrease (approximately 50-90% of control) in [3H]thymidine ([3H]TdR) incorporation. DNA synthesis was inhibited more by confluent than by log phase CM. By day 7, NHEC treated with CM exhibited fewer colonies per culture, fewer cells per colony, and an increased percentage of single cells. Several growth-regulatory gene products found in the nucleus or at the cell membrane have been shown to be expressed differently in normal and transformed cells. We selected the p53 and c-Ha-ras p21 proteins to further investigate the mechanism of alteration of proliferation in cells treated with carcinoma CM. Thus, by day 7, the percentage of NHEC with nuclear localization of wild type p53 (wt p53) was elevated by treatment with CM. In contrast, CM-treated EC cells continued to proliferate, and showed a decrease in the percentage of cells expressing nuclear wt p53 and an increase in the cytoplasmic expression of c-Ha-ras p21. Our studies show that EC cell lines release factors which inhibit the proliferation of NHEC, thus favoring the proliferation of EC cells.

Adenocarcinoma↗
PubMed1995

Transforming growth factor-beta 1 mediates communication between human endometrial carcinoma cells and stromal cells.

The role of transforming growth factor-beta 1 (TGF-beta 1) in communication between human endometrial carcinoma (EC) cells and normal endometrial stromal cells (NSC) was investigated using a cell culture model. Serum-free conditioned medium (CMe) from EC cells (RL95-2, HEC1A) inhibited the proliferation (cells per colony < 50% of control; mitotic index 25-50% of control) of NSC. In contrast, NSC-conditioned medium (CMn) stimulated the proliferation of EC cells, but inhibited the growth of NSC. The proliferation of EC cells was stimulated by the range of dilutions of CMe which inhibited the proliferation of NSC. Using confocal microscopy and a monoclonal antibody, TGF-beta 1, a known product of differentiation in the female reproductive tract, was localized to the cytoplasm of NSC and EC cells. Using a protein slot-blot chemiluminescence method, secreted TGF-beta 1 was detected in serum-free medium conditioned by the growth of NSC and EC cells. TGF-beta 1 antibody-neutralized CMe or CMn stimulated the proliferation of both NSC and EC cells. This study suggests that endometrial carcinoma-stromal cell interactions involve autocrine-paracrine signaling pathways, and that TGF-beta 1 protein is one mediator of such interactions.

Autopsy↗
PubMed1992

Centrifugal separation of carcinoma or atypical cells in voided urine.

A simple density gradient method was used to separate atypical and cancer cells from non-cancer cells in voided urine from patients with transitional cell atypia (moderate and grave atypia) and bladder cancer (squamous cell carcinoma and transitional cell carcinoma). Prior to cell separation, the Saccomanno preserved cells were dispersed by homogenization. After cell separation (5 min x 1400 rpm), atypical and cancer cells were enriched up to 20-fold. Also, most of the leucocytes (68-98%) and squamous cells (47-82%) were absent from density gradient specimen fractions containing the largest percentages of atypical and cancer cells. Peak purity ranges of atypical or cancer cells from different sample classes showed a large degree of overlap. This permitted the pooling of density gradient fractions enriched for atypical or cancer cells, thus increasing the efficiency of the method. Also, following centrifugation, the Papanicolaou-stained specimen fractions showed less background staining than the unprocessed controls, and the cells retained diagnostic morphologic features. We infer that this method may be a useful, low-cost approach for the morphologic study of developing cancers, not only from the urinary bladder, but also from the respiratory tract.

Carcinoma, Squamous Cell↗
PubMed1992

Differential effects of dioctanoylglycerol on fibronectin localization in normal, partially transformed, and malignant human endometrial stromal cells.

In this study, we describe the effects of direct activation of PKC by dioctanoylglycerol (DiC8) on cellular morphology and the localization of fibronectin (Fn) in normal, oncogene-transfected, and malignant human endometrial stromal cells. We questioned whether DiC8, an endogenous specific activator of PKC, would function as a second oncogene in partially transformed human endometrial stromal cells (HESC). Cells utilized were (1) normal HESC, (2) HESC transfected with a plasmid containing an origin-defective temperature-sensitive SV40 large T antigen alone or (3) in combination with an EJ ras oncogene, and (4) an endometrial sarcoma cell line (S7). Cell cultures were treated for 1 h with sn-dioctanoylglycerol (DiC8) and stained with a monoclonal fluorescein-labeled anti-Fn antibody. In normal HESC, DiC8 induced cell rounding and caused Fn localization to revert from the perinuclear region to the cell periphery. All experiments in this investigation were performed when cells were maintained at the permissive temperature for SV40 large T antigen function. In HESC expressing the SV40 large T antigen alone, Fn was localized to the perinuclear region and also occurred as parallel strands between cells. When these cells were treated with DiC8, Fn localization changed to intense punctate regions at the cell periphery or to matrix-like patterns between cells. Also, in these cells, DiC8 induced greater detachment of cells from the substrate than from other cells, resulting in an apparent piling up of cells. Control and treated SV40/EJ ras cells and uterine sarcoma cells expressed Fn in a matrix-like pattern between cells. The rounded cellular morphology of treated HESC and treated cells expressing SV40 resembled the morphology of control or treated SV40/EJ ras cells and uterine sarcoma cells. Thus, treated cells expressing the SV40 large T antigen resembled the SV40/EJ ras cells and uterine sarcoma cells with respect to Fn localization and cellular morphology. DiC8 did not appear to further transform HESC expressing SV40 and EJ ras. However, with regard to cell shape and Fn localization, our results suggest that DiC8 may function as a second oncogene in the signal transduction pathway, in cells expressing SV40 alone. It appears that, with regard to Fn localization, DiC8 may alter signal transduction analogously to that caused by the activated Ha-ras oncogene in HESC expressing the SV40 large T antigen.

Antibodies, Monoclonal↗
PubMed1991

Exfoliative cytologic evaluation of primary cultured lung carcinomas.

This paper describes a tissue culture and exfoliative cell culture system that enables one to (1) evaluate the adequacy of primary lung carcinoma cultures for cytogenetic analysis, and (2) predict the likelihood of viable cells and type of differentiation present in the primary lung tumor cultures used for cytogenetics. Primary lung carcinomas were established from explant outgrowths and maintained in serum supplemented or serum free media on plastic or basement membrane associated protein coated dishes in order to obtain cells for karyotypic analysis (Miura et al., 1990). The media from these cultures that would ordinarily have been discarded was aspirated at each media change and used to prepare cytocentrifuge cytology preparations. Papanicolaou stained cells from the preparations were evaluated by cytotechnologists in order to assess (1) the cellularity and presence of cancer cells in the sample, (2) cytology preparations of cell and explant outgrowth cultures from primary lung tumors are reliable method for screening and evaluating the suitability of primary lung carcinoma cultures for cytogenetic analysis.

Adenocarcinoma↗
PubMed1991

Cell-to-cell communication: a differential response to TGF-beta in normal and transformed (BEAS-2B) human bronchial epithelial cells.

The effects of transforming growth factor beta (TGF-beta) on cell-to-cell communication were investigated in the log phase of growth in normal BE and in adenovirus-12 SV40 hybrid virus transformed BE cells (strain BEAS-2B). Gap junctions in these cells were identified immunocytochemically. Exposure of BE cells to exogenous TGF-beta (0.04-4.0 pM) in serum-free keratinocyte growth medium (KGM) for 1 or 24 h reduced the rate of fluorescent dye transfer (i.e. cell-to-cell communication) by 30-50% in BE cells. Inversely, in BEAS-2B cells, TGF-beta after 1 h induced a 2- to 10-fold increase in the rate of dye transfer. After 24 h of TGF-beta, communication among BEAS-2B cells was not significantly different from controls (no exogenous TGF-beta). The protein kinase C (PKC) inhibitor H-7 induced a dose-dependent enhancement in communication, which was even higher in the presence of TGF-beta (4 pM X 24 h). The calmodulin antagonist W-7 enhanced communication in BEAS-2B cells independently of the presence of TGF-beta. In keratinocyte basal medium (KBM) supplemented with EGF (5 ng/ml) or with TGF-beta (4.0 pM) dye transfer was reduced or enhanced respectively. The combination of EGF and TGF-beta in KBM antagonized the stimulatory effect of the latter on communication in BEAS-2B cells. In BE cells, continuous exposure (4 days) to TGF-beta in KGM induced a dose-dependent inhibition of proliferation and an increased expression of a keratinized, epidermoid phenotype. This correlated with a reduction in the expression of a mucous secretory phenotype. Increased exposure to TGF-beta (0.04-4.0 pM) decreased the labeling index in BEAS-2B cells, but the cells retained a growth advantage over normal BE cells, and did not express a keratinized epidermoid morphology. With respect to dye transfer as an index of cell-to-cell communication, we conclude (i) that an inhibition or enhancement of communication is involved in the response of bronchial epithelial cells to mitogens (e.g. epidermal growth factor) or growth inhibitors (e.g. TGF-beta), (ii) that PKC and Ca(2+)-calmodulin-dependent processes regulate dye transfer, and (iii) the effects of TGF-beta are mediated by PKC.

1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine↗
PubMed1990

Transformed human bronchial epithelial cells (BEAS-2B) alter the growth and morphology of normal human bronchial epithelial cells in vitro.

Normal human bronchial epithelial cells (BE) and adenovirus-12 SV40 hybrid virus transformed, non-tumorigenic human bronchial epithelial cells (BEAS-2B) were cultured for 7 days in a serum-free hormone supplemented medium. BE cells after 3 days in culture were exposed to conditioned medium (CMt) from confluent BEAS-2B cells. By day 7, CMt-treated BE cells exhibited a lower colony forming efficiency (CFE), fewer cells per colony, and a reduced mitotic index (MI) and BrdU (bromodeoxyuridine) labeling index. CMt also enhanced the expression of a terminally differentiated squamous phenotype in BE cells. Cell free lysates from BEAS-2B cells (CFLt) had effects similar to CMt on the MI and morphology of BE cells. In contrast, CMt and CFLt did not inhibit the growth, or alter the morphology of BEAS-2B cells. Conditioned medium from BE cells (CMn) did not reduce the growth of BEAS-2B cells, and had little effect on the morphology of BE cells. In co-cultures, BE cells in direct contact with BEAS-2B cells had a lower MI (0.4-0.7 vs. 1.6%) compared with colonies of BE cells in these cocultures. The concentration of transforming growth factor beta (TGF-beta) in conditioned media from BEAS-2B cells (CMt) was increased 10-fold over that in CMn. TGF-beta is known to induce terminal differentiation in epithelial cells. These results suggest that the selective growth advantage of transformed cells over normal cells during human bronchial carcinogenesis may be related to the release of autocrine/paracrine factors (e.g., TGF-beta) from transformed cells, which down-regulates and terminally differentiates the normal cells.

Bronchi↗
PubMed1990

Intercellular communication in bronchial epithelial cells: review of evidence for a possible role in lung carcinogenesis.

A challenging aspect of lung carcinogenesis is the elucidation of the mechanisms which permit initiated bronchial epithelial cells to attain a growth advantage over normal bronchial epithelial cells, and subsequently evolve into a malignant phenotype. In this review, the effects of interactions between normal and transformed cells, and the potential role of representative extrinsic factors on cell-cell communication are discussed. Evidence is presented to show how cell injury and the effects of serum and calcium may affect morphology and communication, and tumor development. A large number of autocrine-paracrine factors (e.g., TGF beta, TGF alpha) are released by bronchial epithelial cells. These factors may inhibit or promote the proliferation of normal and transformed bronchial epithelial cells, respectively. The ability of certain injurious and tumor promoting agents (e.g., formaldehyde, TPA) to select for the transformed phenotype may involve selective cell injury, the induction of terminal differentiation and an inhibition of gap junction communication among normal BE cells.

Bronchi↗
PubMed1988

Cytomorphologic changes in split-course radiation-treated bronchogenic carcinomas.

The cellular changes produced by radiation therapy were studied in cytologic specimens from seven patients with bronchogenic carcinoma: squamous-cell carcinoma (four patients), adenocarcinoma (two patients), and small-cell undifferentiated carcinoma (one patient). Cytologic samples were obtained from sputum and bronchoscopic examination in patients before and after they received a course of radiation. The cells were studied by light microscopy. An increase in the percentage of columnar cells was detected in five of seven patients independent of the latency of cytologic sampling posttherapy. The percentage of macrophages was unchanged in patients sampled at 15, 18, and 24 mo after the last radiation dose; leukocytes were decreased in relative frequency in the majority of patients studied. Following irradiation, moderately atypical and severely atypical metaplastic cells were increased in relative frequency (48% versus 15% and 19% versus 3%, respectively) and malignant cells were decreased (40% versus 22%) or absent in all patients, irrespective of the type of carcinoma. Nuclear vacuolation, nuclear enlargement, loss of chromatin texture, and rupture of the chromatinic rim were seen in atypical and cancer cells. Infiltration of cancer cells by leukocytes was observed in two of four patients with squamous-cell carcinoma. In all treated patients, morphologic observations indicated that irradiation produces similar damage to "normal" bronchial epithelial cells although such changes are less apparent at longer time intervals following therapy.

Adenocarcinoma↗
PubMed1986

Centrifugal separation of cells in sputum specimens from patients with undifferentiated carcinoma.

Ethanol-fixed cells in sputum from patients with undifferentiated carcinoma of the lung were separated in aqueous Ficoll using a discontinuous density gradient centrifugation technique. The selective enrichment of small cell undifferentiated (e.g., oat cell) or large cell undifferentiated carcinoma cells was achieved while removing most of the leukocytes (80-90%) and macrophages (65-75%) from specimen fractions containing the greatest relative frequencies of cancer cells. The maximum purity of small cell carcinoma cells (0.04%) occurs in moderate density (rho = 1.121 g/ml) gradient fractions and results in a 2.4-fold enrichment relative to unprocessed specimens. In contrast, the maximum purity of large cell carcinoma cells (0.22%) is obtained in very high density (rho = 1.172 g/ml) gradient fractions and results in a 1.2-fold enrichment in comparison with unprocessed specimens. Microscopic examination of Papanicolaou-stained specimen fractions reveals that these enrichments were achieved while retaining diagnostically significant cytomorphologic and tinctorial features necessary for cancer screening and diagnosis. Peak purity ranges of undifferentiated cancer cells significantly overlap comparable ranges for material from bronchogenic adenocarcinoma and squamous cell carcinoma.

Carcinoma↗
PubMed1986

Contact smear cytologic preparations of organ culture explant tissue.

The effects of cytologic sampling by contact-smear on the viability and morphology of epithelial tissues maintained as explants in organ culture were studied. Tracheal and pancreatic tissues from Syrian golden hamsters were maintained for 30 days in culture, and sampled cytologically at weekly intervals. Analysis of sequential Papanicolaou-stained smears demonstrates that explant tissues can be evaluated cytologically over time in vitro without apparent effects on either the tissue viability, histochemistry or morphology. Microscopic examination further reveals that cellular samples retain diagnostically significant cytomorphologic features of the in vivo tissues. These observations indicate that this technique is useful for understanding the long-term response to carcinogens of epithelial tissue maintained in explant organ culture.

Animals↗
PubMed1985

Automatic cell identification and enrichment in lung cancer: V. Adenocarcinoma and large cell undifferentiated carcinoma.

The aims of this study were to develop a protocol for the identification and enrichment of cancer cells from sputum obtained from patients with adenocarcinoma of the lung (n = 6) and large-cell undifferentiated carcinoma of the lung (n = 2), and to compare these findings with the results from our previous studies on other cell types from lung cancer. The hypotheses tested were: Cancer cells in sputum can be preserved following flow sorting. Enrichment for cancer cells from acridine orange (AO)-stained specimens can be achieved. Discrimination of cancer cells from noncancer cells is by AO green fluorescence and discrimination of lymphocytes from other cell types is by AO red fluorescence. Cancer cells are consistently enriched in the AO high green and red fluorescence region, although, for a given cell type, maximal enrichment is patient-dependent. Finally, cancer cell enrichment and lymphocyte exclusion can be done simultaneously. Cells from sputum were initially fixed, stained with AO, sorted on a dual parameter flow sorter, and classified into six groups corresponding to two ranges of green and three ranges of red fluorescence intensities. Cells of each region were stained by the method of Papanicolaou and differential counts were performed to determine the relative frequencies (i.e., purities) of leukocytes, macrophages, squamous cells, and cancer cells, in sorted and unsorted (i.e., control) samples. The average purity of leukocytes (81%), macrophages (6%), squamous cells (11%), and cancer cells (2%) varied markedly from sample to sample. However, the largest enrichment values (i.e., ratio of purity of a cell type in a sorted sample to its purity in the unsorted control sample) achieved for cancer cells consistently occurred for each patient sample in the region corresponding to high green and high red fluorescence intensities. Experimentally, a cancer cell average enrichment of sixteen-fold was obtained by this method. Additionally, fluorescence intensity ranges which increased the enrichment for macrophages by cell sorting typically excluded leukocytes and squamous cells, and vice versa. Finally, red fluorescence intensity was the primary discriminatory parameter for all cell types studied, although the additional use of green fluorescence intensity significantly increased cancer cell enrichment rates.(ABSTRACT TRUNCATED AT 400 WORDS)

Acridine Orange↗
PubMed1981

Centrifugal separation of cells in sputum specimens from patients with adenocarcinoma.

A Ficoll density gradient centrifugation technique was used to separate fixed cells from the sputa of patients with bronchogenic adenocarcinoma. The selective enrichment of leukocytes, squamous or cancerous cells was achieved while retaining the diagnostically significant cell morphologic features. This technique is shown to typically eliminate most of the leukocytes (96.9%) macrophages (86.7%), squamous cells (70.0%) and necrotic debris from specimen fractions containing the majority of cancerous cells. The maximum purity of cancer cells (14.6%) occurs in high density (p = 1.35 g/ml) gradient fractions and results in an average 12.2-fold enrichment for cancer cells in contrast to their relative frequency of occurrence in unprocessed specimens. Differential cell count analyses of serial density centrifugation fractions show that this technique produces comparable enrichment rates for material from bronchogenic adenocarcinoma and squamous cell carcinoma.

Carcinoma, Bronchogenic↗
PubMed1979

Centrifugal separation of cells in sputum.

A centrifugation technique was developed and used to separate fixed cells from the sputa of patients with bronchogenic squamous cell carcinoma and ones with no evidence of cancer. This article presents the relative frequencies of occurrence of five cell types (i.e., leukocytes, macrophages, squamous, columnar and atypical/cancer) in specimen fractions separated from a discontinuous aqueous Ficoll density gradient. These differential counts show that individual cell types may be selectively collected. Atypical and cancer cells are found at high-density gradient fractions (p congruent to 1.138-1.155 g/ml) with a 10-fold enrichment over unprocessed samples.

Carcinoma, Bronchogenic↗
PubMed1979

Automatic cell identification and enrichment in lung cancer. I. Light scatter and fluorescence parameters.

Two physical parameters were investigated to automatically recognize cells in sputum from human squamous cell carcinoma of the lung and to separate them for preparation by the Papanicolaou methods, for human interactive identification and for automated high resolution image analysis. The two parameters, 0.5-15.0 degrees forward argon-ion laser light scatter to estimate total cell size and 546 nm Acridine orange fluorescence to approximate total cell DNA content, were measured in a flow-through fluorescence activated cell sorting system. Enrichment for neoplastic cells in three cases of squamous cell carcinoma of the lung averaged 7.8-fold over the original sputum when only green fluorescence was used and 10.5-fold using green fluorescence and forward light scatter. The average enrichment for neoplastic cells was 65.6-fold relative to polymorphonuclear deenrichment.

Carcinoma, Squamous Cell↗
PubMed1979

Automatic cell identification and enrichment in lung cancer. II. Acridine orange for cell sorting of sputum.

Fluorescence spectra were obtained from cells from sputum and pleural effusions stained with different fluorescent dyes and fixed by alternate methods. The spectra were referenced to a standard allowing for fluorescence comparisons of unstained and stained cells under various conditions. The metachromasia of acridine orange-stained cells offers nuclear/cytoplasmic differentiation in a single stain; mithramycin and propidium iodide do not. Unstained cells have an appreciable amount of green (546 nm) fluorescence, as does Carbowax in Saccomanno's preservative. Cytoplasm stained with acidine orange also has appreciable green fluorescence. Consequently, cells with much cytoplasm have high total fluorescence. Cytoplasmic fluorescence is negligible with mithramycin or propidium iodide. The metachromasia of acridine orange-stained cells is altered by alcohol and Carbowax levels in fixatives, keeping other factors constant.

Acridine Orange↗
PubMed1979

Automatic cell identification and enrichment in lung cancer. III. Light scatter and two fluorescence parameters.

Two fluorescence parameters and size are used in a flow through system to enrich sputum specimens for cancer cells. Human cells in sputum which are stained with acridine orange show a characteristic distribution of red and green fluorescence from which cancer cells can be localized. The peak enrichment is obtained by selectively sorting cells with the largest values of red and green fluorescence. Cancer cells located in other distribution regions having smaller fluorescence intensities show progressively diminished nuclear and cytoplasmic tinctorial features by Papanicolaou stain, consistent with the decreased intensity of red and green fluorescence.

Carcinoma, Squamous Cell↗