Differentiation induction subtraction hybridization (DISH): a strategy for cloning genes displaying differential expression during growth arrest and terminal differentiation

Huang, Fei; Adelman, Jennifer; Jiang, Hai; Goldstein, Neil I.; Fisher, Paul B.

doi:10.1016/s0378-1119(99)00244-9

Cited by 51 publications

(49 citation statements)

References 17 publications

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“…17,2004 In this study, we present evidence from EMSA and Western blot analysis that during melanoma progression the composition and activity of AP-1 dimers are distinctly altered (summary, Table 1). As these studies were done in cell lines derived from melanoma tissues, with the limitations therein, confirmation of these results in primary tissue will be required to confirm their clinical relevance.…”

Section: Primary Metastatic C-junmentioning

confidence: 65%

“…17,2004 Following the instruction manual we also performed nonradioactive EMSA analysis using a DIG Gel shift kit (Roche Molecular Biochemicals, Mannheim, Germany). Briefly, AP-1 oligonuceotides were labeled with digoxigenin-11-ddUTP and incubated with nuclear extract proteins.…”

Section: Nuclear Protein Extractionmentioning

confidence: 99%

“…Following the manufacturer's protocol (Invitrogen, Inc.), mRNA was prepared using a Micro-FastTrack 2.0 kit and the concentration of each sample was determined by spectrophotometry (17). Equal amounts of mRNA (0.1 lg) were amplified in 50 ll reaction volumes containing 1 lM of the appropriate primers (forward primer: 5¢-TCC TAT GAC GAT GCC CTC AAC-3¢; backward primer: 5¢-GTG TTC TGG CTG TGC AGT TC-3¢), 200 lM of each of the dNTPs, 1 ll of RT/Platimum Taq mix and 1· Reaction Mix [SuperScript One-Step RT-PCR with Platinum Taq kit (Invitrogen, Inc.)].…”

Section: Semiquantitative Pcr Amplificationmentioning

confidence: 99%

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During Human Melanoma Progression AP‐1 Binding Pairs are Altered with Loss of c‐Jun In Vitro

Yang

McNulty

Meyskens

2004

Pigment Cell Research

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We demonstrated previously that c‐Jun, JunB and c‐Fos RNA were dysregulated in metastatic melanoma cells compared with normal human melanocytes. The purpose of this study was to evaluate the distribution in composition of AP‐1 dimers in human melanoma pathogenesis. We investigated AP‐1 dimer pairing in radial growth phase‐like (RGP) (w3211) and vertical growth phase‐like (VGP) (w1205) human melanoma cells and metastatic cell lines (cloned from patients, c83‐2c, c81‐46A, A375, respectively) compared with melanocytes using electrophoretic mobility shift assay (EMSA), Western blot and transfection analyses. There are progressive variations in AP‐1 composition in different melanoma cell lines compared with normal melanocytes, in which c‐Jun, JunD and FosB were involved in AP‐1 complexes. In w3211, c‐Jun, JunD and Fra‐1 were involved in AP‐1 binding, while in w1205, overall AP‐1 binding activity was decreased significantly and supershift binding was detected only with JunD antibodies. In metastatic c81‐46A and A375 cells, only JunD was involved in AP‐1 binding activity, but in a third (c83‐2c) c‐Jun, JunD and Fra‐1 were present. Western blot evaluation detected c‐Jun in melanocytes and w3211, but this component was decreased significantly or was not detectable in w1205, c81‐46A and A375 cells. In contrast, JunD protein was elevated in c81‐46A and c83‐2c cells compared with melanocytes and RGP and VGP cell lines. Normal melanocytes and c83‐2c cells (which have c‐Jun involved in AP‐1 binding), transfected with c‐Jun antisense and treated with cisplatin, showed higher viability compared with untransfected cells, while in c81‐46A cells (in which only JunD is detectable) no change in cell viability was observed following treatment with cisplatin and c‐jun antisense transfection. A dominant‐negative c‐Jun mutant (TAM67) significantly increased the soft agar colony formation of w3211 and c83‐2c cells. These results suggest that components of AP‐1, especially c‐Jun, may offer a new target for the prevention or treatment of human melanoma progression.

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Section: Primary Metastatic C-junmentioning

confidence: 65%

Section: Nuclear Protein Extractionmentioning

confidence: 99%

Section: Semiquantitative Pcr Amplificationmentioning

confidence: 99%

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During Human Melanoma Progression AP‐1 Binding Pairs are Altered with Loss of c‐Jun In Vitro

Yang

McNulty

Meyskens

2004

Pigment Cell Research

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“…Selection of subtracted cDNAs was achieved by matching the ends of the cDNA fragments to the ends of the plasmid vectors during ligation and construction of subtracted libraries. Initial screening of a large number of clones representing di erential expression of cellular genes was accomplished by reverse Northern hybridization, followed by Northern blotting to con®rm true di erential expression of genes in infected versus control cells (Huang et al, 1999;Kang et al, 1998). Previous studies document a high degree of conformity (*89%) between reverse Northern and Northern expression of RaSH-derived ESTs (Jiang et al, 2000;Simm et al, 2001).…”

Section: Resultsmentioning

confidence: 99%

“…The PCR products were blotted onto ®lters and reverse Northern blotting was performed to identify cDNAs displaying di erential expression in HIV-1 infected versus uninfected early passage human fetal astrocytes (Huang et al, 1999;Jiang et al, 2000;Kang et al, 1998). cDNAs displaying elevated expression in HIV-1 infected fetal astrocytes versus uninfected fetal astrocytes were designated AEG with a clone number of 1 to 15.…”

Section: Colony Screeningmentioning

confidence: 99%

Identification and cloning of human astrocyte genes displaying elevated expression after infection with HIV-1 or exposure to HIV-1 envelope glycoprotein by rapid subtraction hybridization, RaSH

et al. 2002

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AP-1 and C/EBP transcription factors contribute tomda-7 gene promoter activity during human melanoma differentiation

2000

Self Cite

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Treatment of human melanoma cells with a combination of recombinant fibroblast interferon (IFN-beta) and the protein kinase C (PKC) activator mezerein (MEZ) causes a rapid and irreversible suppression in growth and terminal cell differentiation. Temporal subtraction hybridization combined with random clone selection, reverse Northern hybridization, high throughput microchip cDNA array screening, and serial cDNA library arrays permit the identification and cloning of genes that are differentially expressed during proliferative arrest and terminal differentiation in human melanoma cells. A specific melanoma differentiation associated (mda) gene, mda-7, exhibits reduced expression as a function of melanoma progression from melanocyte to metastatic melanoma. In contrast, treatment of metastatic melanoma cells with IFN-beta + MEZ results in expression of mda-7 mRNA and protein. To evaluate the mechanism underlying the differential expression of mda-7 as a function of melanoma progression and induction of growth arrest and differentiation in human melanoma cells the promoter region of this gene has been isolated from a human placental genomic library and characterized. Sequence analysis by GCG identifies multiple recognition sites for the AP-1 and C/EBP transcription factors. Employing a heterologous mda-7 luciferase gene reporter system, we demonstrate that ectopic expression of either AP-1/cJun or C/EBP can significantly enhance expression of the mda-7 promoter in melanoma cells. In contrast, a dominant negative mutant of cJun, TAM67, is devoid of promoter-enhancing ability. Western blot analyses reveals that cJun and the C/EBP family member C/EBP-beta are physiologically relevant transcription factors whose expression corresponds with mda-7 mRNA expression. Electrophoretic mobility shift assays (EMSA) performed using nuclear protein extracts from terminally differentiated human melanoma cells document binding to regions of the mda-7 promoter that correspond to consensus binding sites for AP-1 and C/EBP. These results provide further mechanistic insights into the regulation of the mda-7 gene during induction of terminal cell differentiation in human melanoma cells.

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Differentiation induction subtraction hybridization (DISH): a strategy for cloning genes displaying differential expression during growth arrest and terminal differentiation

Cited by 51 publications

References 17 publications

During Human Melanoma Progression AP‐1 Binding Pairs are Altered with Loss of c‐Jun In Vitro

During Human Melanoma Progression AP‐1 Binding Pairs are Altered with Loss of c‐Jun In Vitro

Identification and cloning of human astrocyte genes displaying elevated expression after infection with HIV-1 or exposure to HIV-1 envelope glycoprotein by rapid subtraction hybridization, RaSH

AP-1 and C/EBP transcription factors contribute tomda-7 gene promoter activity during human melanoma differentiation

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