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B M Kacinski

Publications and source records attributed to B M Kacinski.

At least 73 records · Page 4Linked to original sources
PubMed1989

Circulating levels of CSF-1 (M-CSF) a lymphohematopoietic cytokine may be a useful marker of disease status in patients with malignant ovarian neoplasms.

We have previously reported our observations of epithelial tumor cell expression of transcripts and protein antigens of CSF-1 and transcripts of a protein closely related--if not identical--to the CSF-1 receptor in ovarian and endometrial neoplasms in vivo and in vitro. In the present communication, we extend these investigations to analyze the clinical utility of determinations of plasma CSF-1 concentrations in 125 samples from 33 patients with ovarian neoplasms and one patient with benign endometriosis. We observed that plasma CSF-1 levels in patients with active and recurrent neoplastic disease were markedly elevated and follow changes in disease status in individual patients. These and other results presented in this communication suggest that not only could CSF-1 be a useful circulating tumor marker in ovarian carcinoma patients, but also that in combination with measurements of other markers--such as CA-125--determinations of plasma CSF-1 levels might actually improve the accuracy of "tumor marker"-based assessments of disease status in patients with malignant ovarian neoplasms.

Adenocarcinoma↗
PubMed1989

Human breast carcinoma cell levels of MDR-1 (P-glycoprotein) transcripts correlate in vivo inversely and reciprocally with tumor progesterone receptor content.

Sixteen human breast carcinomas were subjected to molecular biological and biochemical analyses to determine tumor cell MDR-1 (P-glycoprotein) levels and progesterone receptor content. The results of these analyses disclosed a strong reciprocal and inverse correlation between levels of tumor cell-specific MDR-1 complementary hybrids and progesterone receptor content. These results suggest that the mechanisms which control expression of the P-glycoprotein gene and the progesterone receptor are interrelated and antagonistic, a result with obvious molecular biological, physiological, and clinical implications.

ATP Binding Cassette Transporter, Subfamily B, Mem↗
PubMed1989

In situ transcription and detection of CD1a mRNA in epidermal cells: an alternative to standard in situ hybridization techniques.

To develop methods for the investigation of mRNA transcription in rare epidermal cells, we used in situ transcription to study CD1a mRNA in isolated CD1a positive cells. We chose to study this Langerhans cell marker because it is not known which epidermal cells actually produce the CD1a protein and because there is evidence that CD1a mRNA is alternately spliced, a situation which could lead to truncated or alternate protein products in CD1a surface protein negative cells. Disaggregated epidermal cells were resolved into CD1a surface protein positive and negative groups by fluorescence activated cell sorting and cytocentrifuged onto glass slides. A synthetic 52 base, CD1a specific anti-sense oligomer was hybridized to CD1a gene transcripts in these cells, and radiolabeled cDNA synthesized in situ on the oligomerprimed CD1a transcripts. The labeled cDNA fragments were visualized in the cells of origin by autoradiography, and grains per cell were counted. Sixty-eight percent of cells expressing CD1a protein contained CD1a mRNA, as evidenced by grain counts more than two standard deviations above the mean value for similar cells carried through the same procedure with a control oligomer, or the mean value of CD1a surface protein negative cells treated with the CD1a specific oligomer. Thus, it seems likely that the CD1a protein positive epidermal cells use CD1a mRNA to make their own CD1a protein, and that a truncated or masked CD1a protein is not made by CD1a negative neonatal foreskin epidermal cells. In our hands, in situ transcription is simpler and faster than standard methods of in situ hybridization with prelabeled cDNA or RNA probes. Furthermore, it can be applied to the detection of any message of known sequence. The combination of cell sorting and in situ transcription can be used to localize and quantify the expression of specific mRNA by individual cells, allowing the study of rare and difficult-to-obtain cells.

Antigens, CD1↗
PubMed1989

Oncogene structure, function, and expression in breast cancer.

The research of the past decade has made it clear that aberrant expression of genes that encode growth factors, their receptors, and other genes involved in normal cell growth and differentiation--so-called oncogenes--play important roles in determining the proliferative and invasive characteristics of malignant mammary neoplasms. The authors briefly review the topic of oncogenes and the relevance of oncogenes to breast carcinoma; also included is a description of current techniques commonly employed in the study of aberrant oncogene structure and function. Also presented is the authors' own work on oncogene expression in breast neoplasms, which implicates the neu, fms, PDGF-A and Ki-ras oncogenes in the control of mammary epithelial cell proliferation by autocrine and paracrine mechanisms.

Breast Neoplasms↗
PubMed1988

High level expression of fms proto-oncogene mRNA is observed in clinically aggressive human endometrial adenocarcinomas.

Six micron paraffin sections of paraformaldehyde-fixed endometrial currettings of 21 benign and neoplastic endometrial specimens were assayed for tumor cell-specific oncogene expression by in situ hybridization with probes for six oncogenes, beta-actin, and the E. coli plasmid pBR322. In the benign hyperplasias and invasive adenocarcinomas, multiple oncogenes, including erbB, fms, c-myc, and Ki-ras were expressed at significant levels. For the adenocarcinomas, statistical analysis demonstrated that high levels of expression of fms-complementary mRNA correlated strongly with clinicopathologic features (high FIGO histologic grade, high FIGO clinical stage, deep myometrial penetration) predictive of aggressive clinical behavior and poor outcome. The authors discuss the role which M-CSF receptor (the fms gene product) and locally-produced M-CSF may play in the development of the observed aggressively-malignant phenotypes. They also propose that pre-hysterectomy assay of fms gene expression in endometrial currettings in FIGO Stage I patients might be clinically useful to help identify preoperatively those patients with deep myometrial penetration or other locoregional spread.

Adenocarcinoma↗
PubMed1987

Therapeutic concepts of brachytherapy/megavoltage in sequence for pharyngeal wall cancers. Results of integrated dose therapy.

This is a follow-up report on lateral, posterior oro-, hypopharyngeal wall cancer treatment based on a new therapeutic concept. Ir-192 or I-125 brachytherapy and craniocervical megavoltage irradiation are effective with improved local control and NED (no evidence of disease) survival rates. Local failure rate is 14% and actuarial survival (NED) is 82% at 5 years in 14 patients treated thus far.

Actuarial Analysis↗
PubMed1986

Demonstration of myc and ras oncogene expression by hybridization in situ in hydatidiform mole and in the BeWo choriocarcinoma cell line.

With the use of hybridization in situ, c-myc and c-ras oncogene expression has been identified in the cytotrophoblast of hydatidiform mole and in the malignant trophoblast cell line BeWo. Expression was also found in early villous placenta. No expression of these two oncogenes was found in the cytotrophoblast of an 11-week conceptus, nor was it found in term placenta. The significance of these findings is discussed.

Cell Line↗
PubMed1985

Iodine-125 implants for carcinoma of the prostate.

One hundred-thirteen patients underwent Iodine-125 prostate implant and lymphadenectomy at Yale-New Haven Hospital from 1974 through 1980. The distribution by clinical stage was: 7 Stage A2, 86 Stage B, and 20 Stage C patients. Ninety-four patients had a negative lymphadenectomy (N-) and 19 patients (17%) had metastatic disease in the pelvic lymph nodes (N+). The actuarial 5-year survival for all 113 patients was 87% (+/- 6%: 95% confidence limits). Sixty-five percent of our 113 patients are disease free (NED) from 2 to 9 years following implant. Sixty-seven (N-) patients with clinical Stage B disease, whose tumors were either well differentiated or moderately well differentiated, have an actuarial 5-year NED survival of 84% (+/- 8%). Twenty (N-) patients with either clinical Stage C disease or poorly differentiated tumors have an actuarial 5-year NED survival of only 31% (+/- 20%). For the 19 (N+) patients, the actuarial 5-year NED survival is 38% (+/- 22%). Local tumor control was 85% for all Stage B patients and 75% for all Stage C patients. Only 10 patients (9%) have developed long-term gastrointestinal or genitourinary complications following their implant. Iodine-125 implant appears to be a reasonable alternate form of therapy in highly selected groups of patients with carcinoma of the prostate.

Adenocarcinoma↗
PubMed1985

Repair of haloethylnitrosourea-induced DNA damage in mutant and adapted bacteria.

The sensitivities of Escherichia coli K-12 strain AB1157, its uvrA-deficient mutant AB1886, and its recA mutant AB2463 to N,N'-bis(2-chloroethyl)-N-nitrosourea, N-(2-chloroethyl)-N-nitrosourea, and N-ethyl-N-nitrosourea have been determined. These data indicate that loss of either uvr excision repair or recA-dependent DNA repair greatly increases sensitivity to the haloethylnitrosoureas. At the same time, loss of recA-dependent DNA repair increases sensitivity to N-ethyl-N-nitrosourea significantly while loss of uvr excision repair increases sensitivity to this agent only marginally. Adapting the uvrA-deficient and recA-deficient mutants by growth in N-methyl-N'-nitro-N-nitrosoguanidine increases survival after exposure to either N-methyl-N'-nitro-N-nitrosoguanidine or N-ethyl-N-nitrosourea, but neither adapted strain loses its sensitivity to N,N'-bis(2-chloroethyl)-N-nitrosourea. Taken together, these data indicate that the haloethylnitrosoureas cause other important cytotoxic lesions in DNA in addition to those involving alkylation of the O6 position of guanine and that the uvrA and recA gene products are involved in the repair of these lesions.

Alkylation↗
PubMed1984

Interactions of the UVRABC endonuclease in vivo and in vitro with DNA damage produced by antineoplastic anthracyclines.

The anthracycline antineoplastic agents Adriamycin and N-trifluoroacetyl-Adriamycin-14-valerate were assayed in vivo and in vitro for ability to produce DNA lesions recognized by the UVRABC endonuclease, a DNA repair enzyme of Escherichia coli which recognizes large, bulky lesions in DNA. We found that, while both drugs produce DNA lesions, only the lesions produced by Adriamycin were toxic. Hence, anthracycline antineoplastic activity may be related to production of large, bulky lesions in DNA, while toxicity may correlate with toxicity measured in a simple E. coli DNA repair mutant test system.

DNA Repair↗
PubMed1981

A general approach for purifying proteins encoded by cloned genes without using a functional assay: isolation of the uvrA gene product from radiolabeled maxicells.

The uvrA protein (UVRA) of E. coli has been extensively purified from a strain in which UVRA is overproduced and specifically labeled with 35S-methionine. This approximately 100-fold overproduction relative to normal strains is a result of having the uvrA gene present on a multicopy plasmid in a spr recA cell that makes defective lexA protein, the normal repressor of the uvrA gene, while the specific labeling of UVRA is done with maxicells. This approach facilitates the preparation of the protein since enzyme assays do not have to be carried out during the intermediate stages of purification. The purified UVRA binds to DNA and has ATPase activity but does not have intrinsic endonuclease activity. When added to extracts of uvrA- cells, the purified UVRA does promote the specific cutting of UV-irradiated DNA. Since this approach for working out rapid purification procedures by specifically labeling the proteins encoded by cloned genes does not require the use of a functional assay, it is a general one that can be applied to a wide variety of other gene products in addition to UVRA.

Adenosine Triphosphatases↗
PubMed1981

Identification of the uvrC gene product.

We have constructed a multicopy plasmid that carries the uvrC gene of Escherichia coli. By inserting the transposon Tn1000 (previously designated gamma delta) into this plasmid, we obtained many derivatives that fail to complement uvrC34. The proteins synthesized by the original plasmid and the uvrC::Tn1000 derivatives were labeled in maxicells and analyzed on gels, demonstrating that a protein of Mr 70,000 encoded by the original uvrC+ plasmid was absent from the mutated noncomplementing derivatives; this protein is presumed to be the uvrC gene product. We found that this protein of Mr 70,000 binds to DNA and have partially purified the uvrC protein by DNA-cellulose chromatography. Because some of the uvrC::Tn1000 derivatives produce truncated polypeptides, the orientation of expression and the location of the promoter were determined by correlating the sizes of the truncated polypeptides with the sites of insertion of Tn1000.

Bacterial Proteins↗
PubMed1980

Effect of excision repair on the mutagenesis of misonidazole.

The mutagenic and toxic effects of the nitroimidazole hypoxic cell sensitizer misonidazole (Ro-07-0582) have been studied in two strains of E. coli that differ in their abilities to repair DNA. It was found that even though the strain defective in excision repair was only slightly more sensitive than the original wild-type strain to killing by misonidazole, a large difference in the mutagenesis of the two strains was observed. While the wild-type strain was not mutated by misonidazole at any of the concentrations tested, the uvr strain was strongly mutated by the drug. This demonstrates the presence of a mutagenic lesion in DNA that is removed by excision repair.

DNA Repair↗
PubMedDate not supplied

Expression of CSF-I and CSF-I receptor by normal lactating mammary epithelial cells.

OBJECTIVE: Previous studies suggested a potential role for macrophage colony stimulating factor (CSF-1) in lactogenic differentiation of the breast. The aim of this study was to define the regulation of CSF-1 and its receptor (CSF-1R, the product of c-fms proto-oncogene) by lactogenic hormones in the breast in vivo during pregnancy and lactation and in vitro in organ culture and mammary epithelial cell lines. METHODS: Immunohistochemical staining assays for the expression of CSF-1 and CSF-1R antigens were performed on sections of breast biopsies from nonpregnant (n = 10), prepartum (n = 4), and postpartum lactating patients (n = 7) and on sections of human mammary glands cultured in the presence of the lactogenic hormones insulin, prolactin, and glucocorticoids. Northern blot analyses were used to study the regulation of CSF-1 and CSF-1R by these same lactogenic hormones in normal and neoplastic mammary epithelial cell lines in cell culture. RESULTS: Normal, nonlactating mammary epithelium did not express CSF-1R and synthesized only low levels of CSF-1. During lactation, significant levels of both proteins could be observed in the epithelial cells that line actively lactating ducts and alveoli. Very similar increases in epithelial cell expression of CSF-1 and CSF-1R were observed in organ cultures of normal mammary gland biopsies exposed to prolactin, insulin, and glucocorticoids. Colony stimulating factor mRNA levels were increased by prolactin and/or insulin in a normal mammary epithelial cell line, while glucocorticoids had no apparent effect on CSF-1 mRNA levels. In contrast, we found that the levels of CSF-1R transcript are regulated primarily by glucocorticoids in breast carcinoma cells, while prolactin merely modulates the glucocorticoid effect. CONCLUSION: The observed increases in the expression of CSF-1 and its receptor during lactogenesis and the regulation of CSF-1/CSF-1R by lactogenic hormones suggests that this cytokine/receptor pair might play a regulatory role in the cellular events leading to the lactogenic differentiation of mammary epithelial cells.

Animals↗
PubMedDate not supplied

Messenger RNA decay of macrophage colony-stimulating factor in human ovarian carcinomas in vitro.

OBJECTIVE: Recently, the importance of the macrophage colony-stimulating factor (CSF-1) and its receptor (encoded by the c-fms proto-oncogene) in patients with epithelial ovarian cancer has been recognized. Overexpression of CSF-1 denotes poor prognosis. Macrophage colony-stimulating factor may be one of a group of inflammatory cytokines, whose 3' untranslated region (UTR) contains AU-rich stretches and whose expression may be largely dependent on mRNA decay. The purposes of this study were to investigate the effect of protein synthesis inhibition on CSF-1 transcript expression, to help determine whether there is a role for labile intermediary proteins in the regulation of CSF-1 expression, and to explore the transcriptional or post-transcriptional mechanisms that could underlie such overexpression of CSF-1 transcripts by protein synthesis inhibitors. Although regulation of CSF-1 gene expression has been investigated in hematopoietic cells, such studies have not been carried out in any epithelial cancer. METHODS: Northern blot analyses for CSF-1 expression were performed on total cellular RNA extracted from primary and established ovarian cancer cell lines in the absence or presence of proteins synthesis inhibitors with different modes of action. The probe for the AU-rich exon 10 of CSF-1 was prepared by amplification of the terminal 143 bp of the 3' UTR of human CSF-1 by polymerase chain reaction. Transcription rates were assessed in the presence or absence of cycloheximide (CH) in nuclei of ovarian cancer cells by run-off analyses. The effect of CH on CSF-1 mRNA half-life was measured by actinomycin D chase experiments in SKOV3 cells. RESULTS: We demonstrate that CSF-1 mRNA was expressed by all of a panel of primary and established ovarian cancer cell lines. There are at least three CSF-1 transcripts expressed by ovarian cancer cells; we demonstrate that the 4.2-kb CSF-1 transcript contains exon-6 sequences, which are spliced from the 3.4- and 1.9-kb transcripts, whereas both the 4.2- and 3.4-kb CSF-1 transcripts contain the 3' AU-rich exon 10. Treatment with several different protein synthesis inhibitors resulted in marked overexpression of CSF-1 transcript levels, suggesting a potential role for labile proteins in the regulation of CSF-1 expression. The predominant effect of CH is on the two CSF-1 transcripts that contain exon 10. Although CH does not change the rate of CSF-1 gene transcription, measurement of CSF-1 mRNA stability reveals a prolongation of CSF-1 transcript half-life by CH from 4.5 hours to significantly greater than 6 hours in ovarian cancer cells. CONCLUSIONS: Augmented CSF-1 transcript stability underlies the marked overexpression of CSF-1 seen with protein synthesis inhibitors. Our results suggest the involvement of a labile regulatory protein that contributes to CSF-1 mRNA decay in ovarian cancer cells. Our data suggest further that exon 10 (not the spliced exon-6 sequences) of the CSF-1 transcript may contain an instability determinant.

Cycloheximide↗