1: Mol Med. 2000 Sep;6(9):750-65. Changes in tumorigenesis- and angiogenesis-related gene transcript abundance profiles in ovarian cancer detected by tailored high density cDNA arrays. Martoglio AM, Tom BD, Starkey M, Corps AN, Charnock-Jones DS, Smith SK. Department of Obstetrics and Gynaecology, University of Cambridge, England. amm53@hermes.cam.ac.uk BACKGROUND: Complementary DNA array analysis of gene expression has a potential application for clinical diagnosis of disease processes. However, accessibility, affordability, reproducibility of results, and management of the data generated remain issues of concern. Use of cDNA arrays tailored for studies of specific pathways, tissues, or disease states may render a cost- and time-effective method to define potential hallmark genotype alterations. MATERIALS AND METHODS: We produced a 332-membered human cDNA array on nylon membranes tailored for studies of angiogenesis and tumorigenesis in reproductive disease. We tested the system for reproducibility using a novel statistical approach for analysis of array data and employed the arrays to investigate gene expression alterations in ovarian cancer. RESULTS: Intra-assay analysis and removal of agreement outliers was shown to be a critical step prior to interpretation of cDNA array data. The system revealed highly reproducible results, with intermembrane coefficient of reproducibility of +/- 0.98. Comparison of placental and ovarian sample data confirmed expected differences in angiogenic profiles and tissue-specific markers, such as human placental lactogen (hPL). Analysis of expression profiles of five normal ovary and four poorly differentiated serous papillary ovarian adenocarcinoma samples revealed an overall increase in angiogenesis-related markers, including vascular endothelial growth factor (VEGF) and angiopoietin-1 in the diseased tissue. These were accompanied by increases in immune response mediators (e.g. HLA-DR, Ron), apoptotic and neoplastic markers (e.g. BAD protein, b-myb), and novel potential markers of ovarian cancer, such as cofilin, moesin, and neuron-restrictive silencer factor (REST) protein. CONCLUSIONS: In-house production of tailored cDNA arrays, coupled to comprehensive analysis of resulting hybridization profiles, provides an accessible, reliable, and highly effective method of applying array technology to study disease processes. In the ovary, abundance of specific tumor markers, increased macrophage recruitment mediators, a late-stage angiogenesis profile, and the presence of chemoresistance-related markers distinguished normal and advanced ovarian cancer tissue samples. Detection of such parallel changes in pathway- and tissue-specific markers may prove a hallmark ready for application in reproductive disease diagnostic and therapeutic developments. PMID: 11071270 [PubMed - indexed for MEDLINE] --------------------------------------------------------------- 2: Indian J Exp Biol. 1998 May;36(5):447-55. Oncogene expression as detected by immunocytochemical staining in hormonally induced ovarian cell lines. Luthra K, Chapekar TN. Department of Biochemistry, All India Institute of Medical Sciences, New Delhi, India. To understand the nature and extent of oncogene involvement in the development of neoplasia, an experimental model of goat ovarian granulosa cells stimulated by LH was chosen. In the course of these studies, several cell lines were developed which were essentially non-tumorigenic primary cell lines. One of them, however, was spontaneously transformed being immortalized and tumorigenic. These cell lines, transformed and non-transformed, should serve as contralateral cell lines to study differential oncogene expression in hormonally induced cell proliferation, and elucidate possible hormone-oncogene nexus which may be operative in the genesis of cancer. In the present report, we have studied expression of c-myc, c-ras, c-myb, c-fos and c-sis cellular oncogenes in the cell lines by immunocytochemistry using monoclonal antibodies. In the rest of our text we refer to these cellular oncogenes as oncogenes. The results reveal differential expression of the oncogenes. The striking difference between the non-transformed AIMS/GRXII cells and the transformed AIMS/GRXVIII cells was the absence of ras protein expression in the transformed AIMS/GRXVIII cells which intensely expressed the c-myc, c-myb, c-fos, and c-sis proteins. c-ras protein was expressed in the non-transformed AIMS/GRXVIII cell line and primary cultures. c-myc protein was expressed exclusively in the AIMS/GRXVIII transformed cells. The myc activity seen in the transformed cell line may be correlated to cell proliferation. These results show the variation of phenotype in cell lines derived from a single tissue source. PMID: 9717461 [PubMed - indexed for MEDLINE] --------------------------------------------------------------- 3: Leuk Lymphoma. 1997 Jul;26(3-4):271-9. The A-myb transcription factor in neoplastic and normal B cells. Golay J, Facchinetti V, Ying G, Introna M. Department of Immunology and Cellular Biology, Istituto Ricerche Farmacologiche Mario Negri, Milan, Italy. The myb family of transcription factors has been strongly implicated in the regulation of cell growth and differentiation in the haematopoietic system. The v-myb oncogene, carried by avian defective retroviruses, causes leukaemias in the chicken and transforms haematopoietic cells in vitro. Its normal cellular equivalent c-myb, has been shown to promote the proliferation and block the differentiation of haematopoietic cells in several experimental models and is required for fetal haematopoiesis. Two other members of the family have been cloned more recently, A-myb and B-myb, which show sequence homology with c-myb in several domains, of which the DNA binding domain as well as other regulatory domains. Both have been shown to be transcription factors. B-myb is also involved in the control of proliferation and differentiation, but, unlike c-myb, it is expressed in many cell types. The third member of the family, A-myb, shows the most restricted pattern of expression, suggesting a very specific role for this transcription factor. A-myb is expressed in a subpopulation of normal B lymphocytes activated in vivo and localised in the germinal center of peripheral lymphoid organs and is not detected at significant levels in all other mature or immature haematopoietic populations studied, including bone marrow cells, T lymphocytes, granulocytes, monocytes, either at rest or after in vitro activation. These studies indicate that A-myb plays a role during a narrow window of normal B cell differentiation. A-myb expression has also been studied in a wide range of neoplastic B cells, representing the whole spectrum of B cell differentiation. A-myb is strongly expressed in Burkitt's lymphomas (BL) and slg+ B-acute lymphoblastic leukaemias (B-ALL) and not in all other leukaemias/lymphomas tested, with the exception of a subset of CLL (about 25% of cases). It is intriguing that the A-myb genome has been localised relatively close to the c-myc gene on chromosome 8, suggesting that the c-myc translocation in BL and B-ALL may affect A-myb transcription. Studies are in progress to investigate the functional relationship between A-myb and c-myc, particularly in the context of BL cells and to determine whether A-myb is deregulated in these cells. Publication Types: Review Review, Tutorial PMID: 9322889 [PubMed - indexed for MEDLINE] --------------------------------------------------------------- 4: Plant Mol Biol. 1996 Nov;32(4):707-16. A family of novel myb-related genes from the resurrection plant Craterostigma plantagineum are specifically expressed in callus and roots in response to ABA or desiccation. Iturriaga G, Leyns L, Villegas A, Gharaibeh R, Salamini F, Bartels D. Instituto de Biotecnologia-UNAM, Av. Universidad 2001, Col. Chamilpa, Mexico. A cDNA and two genomic clones comprising highly similar genes that encode a protein with a Myb-related DNA-binding domain were isolated from the resurrection plant Craterostigma plantagineum. The structure of cpm5 and cpm10 (Craterostigma plantagineum myb) genes consists of three putative exons encoding a protein of 36.6 kDa. The cDNA of cpm7 encodes a closely related protein of 36.8 kDa. The canonical Myb domain present in transcriptional activators of yeast, animals and plants was localized in the amino terminus of deduced Cpm5, Cpm7 and Cpm10 proteins and corresponds to the two Myb repeats found in plants. The Myb domain of Cpm deduced proteins and a short stretch of amino acids adjacent to this region are closely related to a myb gene from Arabidopsis thaliana which is expressed in response to osmotic stress and ABA. The rest of the deduced protein has no similarity to other reported sequences. The myb-related genes in the Craterostigma genome comprise a small gene family of 6-8 members as estimated by hybridization with a bona fide Myb domain probe. Northern blot experiments showed specific expression of cpm10 in undifferentiated callus tissue up-modulated by ABA and expression of cpm7 mRNA in roots up-regulated by dehydration. PMID: 8980522 [PubMed - indexed for MEDLINE] --------------------------------------------------------------- 5: J Invest Dermatol. 1995 Jan;104(1):78-82. Changes in expression of apoptosis-associated genes in skin mark early catagen. Seiberg M, Marthinuss J, Stenn KS. Skin Biology Research Center, R.W. Johnson Pharmaceutical Research Institute, Raritan, NJ 08869-0602. Programmed cell death is central to hair biology, as the hair follicle undergoes cycles of growth (anagen), regression (catagen), and rest (telogen). During catagen, the hair follicle shortens via a pathway of programmed cell death and apoptosis. The molecular mechanisms involved in this process have not been elucidated yet. Using reverse transcriptase-polymerase chain reaction, we examined in this study the expression in total skin, throughout one hair cycle, of a series of regulatory genes associated with apoptosis. We show that gene expression within skin is hair-cycle-dependent. Transforming growth factor-beta was expressed immediately before catagen; therefore, it might be involved in the early signaling of this process. Tumor necrosis factor-beta was expressed during catagen and might be involved in follicular apoptosis. Several proto-oncogenes and transcription factors have been described in the regulation of apoptosis in other systems. Here we show that the transcript levels of c-myc, c-myb, and c-jun changed immediately before or during early catagen and thus could be involved in the signaling or regulation of catagen. Levels of p53 remained constant throughout anagen and catagen, suggesting that p53 is not involved in the developmentally induced apoptosis of the hair follicle. The variable expression throughout the hair cycle of the genes described demonstrates the dynamic changes of the skin and underscores the importance of studying the complete hair cycle when characterizing any molecule in skin. PMID: 7798646 [PubMed - indexed for MEDLINE] --------------------------------------------------------------- 6: J Immunol. 1994 Jul 15;153(2):543-53. The A-myb gene is preferentially expressed in tonsillar CD38+, CD39-, and sIgM- B lymphocytes and in Burkitt's lymphoma cell lines. Golay J, Erba E, Bernasconi S, Peri G, Introna M. Institute of Pharmacological Research Mario Negri, Milan, Italy. The A-myb gene is structurally related to the c-mby proto-oncogene, a transcription factor involved in the regulation of hemopoietic proliferation and differentiation. Recent evidence has shown that A-myb also functions as a transcriptional activator. We have previously demonstrated that A-myb RNA is not expressed in most mature human leukocytes at rest or after mitogenic or functional activation. We show here, by using cell sorting, PCR, and Western analyses that A-myb is most highly expressed in the subsets of human tonsillar B lymphocytes with the phenotypes CD38+, CD39-, and SIgM-. The preferential expression of A-myb in these populations was seen at both the RNA and protein levels. CD38 was consistently best at separating high from low A-myb-expressing cells, whereas other markers (CD10, 22, 23, 77, 11a, and 49d) did not correlate with A-myb expression. The CD38+ population expressing the highest levels of A-myb was shown to contain mostly cycling cells inasmuch as more than 95% were in the late G1, S, G2, and M phases of the cell cycle. In addition, A-myb expression always correlated with the percentage of cells in S/G2/M in the populations sorted with either CD38, CD39, or sIgM. Small resting tonsillar B lymphocytes induced to proliferate in vitro by several different polyclonal B cell activators did not, however, express detectable levels of A-myb, although these cells were demonstrated to express CD38 and enter the S/G2/M phases of the cell cycle. These data suggest that A-myb is a marker of in vivo-activated but not in vitro-activated B lymphocytes. Finally, A-myb was also found to be highly expressed in five of seven Burkitt's lymphoma lines and in none of three EBV lymphoblastoid cell lines. This finding is in agreement with the phenotype of the normal B cells that express high levels of A-myb in vivo and suggests that A-myb may be specifically induced within germinal center B cells. PMID: 8021494 [PubMed - indexed for MEDLINE] --------------------------------------------------------------- 7: Zhonghua Zhong Liu Za Zhi. 1991 Jul;13(4):261-4. [Expression of protooncogenes c-myb and c-myc during the hexamethylene bisacetamide (HMBA)-induced commitment to terminal differentiation of murine erythroleukemia cells] [Article in Chinese] Chen ZX. Jiangsu Institute of Hematology, Suzhou. Modulation of a number of protooncogene expression occurs during differentiation of eukaryotes. Changes in expression of c-myb and c-myc during the HMBA-induced terminal differentiation of murine erythroleukemia cells were characterized by an early decrease (within 4 hrs), followed by the recovery of c-myc mRNA by 10 hrs, and the retention of suppression of c-myb expression for the rest of the induction period. Two MELC variants were used to further define the relationship between the differentiation and the protooncogene expression. R1 was a MELC variant completely resistant to HMBA, and R1 (VCR), a vincristine resistant R1, became inducible by HMBA again. The cell differentiation and the c-myb and c-myc expression were determined on R1 or R1 (VCR) cultured with HMBA respectively. The results demonstrated that the c-myc mRNA increased and remained relatively high as the cells grew to a saturated density regardless of the induction of differentiation. The R1 (VCR) cultured with HMBA displayed an early decrease in c-myb mRNA and a subsequent suppression of its expression, while the R1 cultured with HMBA showed a stable level of c-myb mRNA. These results suggest that the c-myb expression, rather than c-myc expression, is closely related to the HMBA-induced terminal differentiation. PMID: 1806345 [PubMed - indexed for MEDLINE] --------------------------------------------------------------- 8: Biull Eksp Biol Med. 1988 Mar;105(3):326-9. [Cellular oncogene expression in human tumors transplantable to athymic mice] [Article in Russian] Spitkovskii DD, Revazova ES. Expression of 3 cellular oncogenes among 7 ones under investigation is identified in the majority of 20 strains of human tumors, passaged in nude mice without significant specificity as far as the type of the tumor is concerned. The levels of the expression of these 3 oncogenes (c-myc, c-fos, c-ras) were higher than the ones in primary human tumors except for the human melanoma Mel-2 strain, where the expression of c-myb oncogene was identified. All the rest oncogenes (c-mos, B-lym, c-sis, c-myb) showed no expression in human tumors of the examined strains. PMID: 3349173 [PubMed - indexed for MEDLINE] ---------------------------------------------------------------