V-myb Myeloblastosis Viral Oncogene Homolog Research Articles

MaMYBR30, a Novel 1R-MYB, Plays Important Roles in Plant Development and Abiotic Stress Resistance.

The V-myb myeloblastosis viral oncogene homolog (MYB) family participate in various bioprocesses including development and abiotic stress responses. In the present study, we first report a 1R SHAQKYF-class MYB, MaMYBR30, in mulberry. Subcellular localization and sequence analysis indicated MaMYBR30 is located in the nucleus and belongs to a CCA-like subgroup with a conserved SHAQKYF motif. Expression profile analysis showed that MaMYBR30 is expressed in leaves and can be induced by drought and salt stress. The down-regulation of MaMYBR30 using virus-induced gene silence (VIGS) in mulberry and the overexpression of MaMYBR30 in Arabidopsis were induced to explore the function of MaMYBR30. The functional characterization of MaMYBR30 in vivo indicated that MaMYBR30 can positively regulate the resistance of mulberry to drought while negatively regulating the resistance of mulberry to salt stress. In addition, MaMYBR30 also affects flower development and reproductive growth, especially after exposure to salt stress. Weighted gene co-expression network analysis (WGCNA) primarily revealed the possible genes and signal pathways that are regulated by MaMYBR30. Our results also imply that complex molecular mechanisms mediated by MaMYBR30, including crosstalk of ion toxicity, phytohormone signal transduction, flowering development, and epigenetic modification, need to be further explored in the future.

Comprehensive analysis of the MYB transcription factor gene family in Morus alba

BackgroundThe V-myb myeloblastosis viral oncogene homolog (MYB) family of proteins is large, containing functionally diverse transcription factors. However, MYBs in Morus are still poorly annotated and a comprehensive functional analysis of these transcription factors is lacking.ResultsIn the present study, a genome-wide identification of MYBs in Morus alba was performed. In total 166 MaMYBs were identified, including 103 R2R3-MYBs and four 3R-MaMYBs. Comprehensive analyses, including the phylogenetic analysis with putative functional annotation, motif and structure analysis, gene structure organization, promoter analysis, chromosomal localization, and syntenic relationships of R2R3-MaMYBs and 3R-MaMYBs, provided primary characterization for these MaMYBs. R2R3-MaMYBs covered the subgroups reported for R2R3-MYBs in Arabidopsis and Populus, and had two Morus-specific subgroups, indicating the high retention of MYBs in Morus. Motif analysis revealed high conservative residues at the start and end of each helix and residues consisting of the third helix in R2 and R3 repeats. Thirteen intron/exon patterns (a–m) were summarized, and the intron/exon pattern of two introns with phase numbers of 0 and 2 was the prevalent pattern for R2R3-MaMYBs. Various cis-elements in promoter regions were identified, and were mainly related to light response, development, phytohormone response, and abiotic and biotic stress response and secondary metabolite production. Expression patterns of R2R3-MaMYBs in different organs showed that MaMYBs involved in secondary cell wall components and stress responsiveness were preferentially expressed in roots or stems. R2R3-MaMYBs involved in flavonoid biosynthesis and anthocyanin accumulation were identified and characterized based on functional annotation and correlation of their expression levels with anthocyanin contents.ConclusionBased on a comprehensive analysis, this work provided functional annotation for R2R3-MYBs and an informative reference for further functional dissection of MYBs in Morus.Supplementary InformationThe online version contains supplementary material available at 10.1186/s12870-022-03626-5.

The Physiological and Molecular Mechanism of Abscisic Acid in Regulation of Fleshy Fruit Ripening.

The ripening of fleshy fruits is coupled with the degradation of both chlorophyll and cell walls, as well as changes in the metabolism of phenylpropanoids, flavonoids, starch/sucrose, and carotenoids. These processes are controlled by phytohormones and other factors, including abscisic acid (ABA), ethylene, auxin, polyamines, sugar, and reactive oxygen species. The ripening of climacteric fruits is controlled by ethylene and non-climacteric fruit ripening is regulated mainly by ABA. Also, ABA and ethylene may interact in both types of fruit ripening. ABA concentrations in fleshy fruits are regulated in response to developmental and environmental cues and are controlled by the relative rates of ABA biosynthesis and catabolism, the former mainly via 9-cis-epoxycarotenoid dioxygenases (NCEDs) and β-glucosidases and the latter via ABA 8'-hydroxylases (CYP707As) and β-glycosyltransferases. In strawberry fruit ripening, ABA is perceived via at least two receptors, Pyrabactin resistance (PYR)/PYR-like (PYL) and putative abscisic acid receptor (ABAR), which are linked separately to the conserved signaling pathway ABA-FaPYR1-FaABIl-FaSnRK2 and the novel signaling pathway ABA-FaABAR-FaRIPK1-FaABI4. Downstream signaling components include important transcription factors, such as AREB (ABA responsive element binding protein)/ABF (ABRE binding factors ABA responsive factor), ethylene response factor (ERF), and V-myb Myeloblastosis viral oncogene homolog (MYB), as well as ripening-related genes. Finally, a comprehensive model of ABA linked to ethylene, sugar, polyamines, auxin and reactive oxygen species in the regulation of strawberry fruit ripening is proposed. Next, new integrated mechanisms, including two ABA signaling pathways, ABA and ethylene signaling pathways, and ABA/ethylene to other phytohormones are interesting and important research topics in ripening, especially in non-climacteric fruits.

Alterations in the silymarin metabolism in transgenic Silybum marianum cultured cells by the heterologous expression of the Arabidopsis thaliana V-myb myeloblastosis viral oncogene homolog transcription factor MYB12 and Cicer arietinum chalcone synthase

Silymarin (Sm), the main bioactive principle of Silybum marianum (L.) Gaernt. fruits, has strong antihepatotoxic properties and also exhibits antitumor and antiviral activity. This flavonolignan complex is synthesized from the flavonoid precursor taxifolin, which undergoes an oxidative coupling with the monolignol coniferyl alcohol. Attempts to produce Sm in plant cell suspensions have met with limited success, probably due to insufficient flux into the flavonoid branch pathway of Sm biosynthesis.In this study, it was attempted to increase Sm production in in vitro cultures of S. marianum by genetically engineering the flavonoid pathway. Using Agrobacterium tumefaciens strain C58C1, S. marianum cell suspension cultures were transformed either with the Arabidopsis thaliana transcription factor AtMYB12 (V-myb myeloblastosis viral oncogene homolog) (MYB) or with a chalcone synthase (CHS) gene from Cicer arietinum. A double transformant was also generated by introducing the CaCHS gene in AtMYB12-expressing cultures.The ectopic expression of AtMYB12 in cultures activated some of the genes involved in the flavonoid pathway, notably chalcone isomerase (CHI) and flavonoid 3′-hydroxylase (F3’H), whereas the secondary metabolites altered by the inserted gene were chlorogenic acid glycoside derivatives. Expression of CaCHS in the cultures led to the production of the flavanone naringenin (Ng), which accumulated as glycosides. The simultaneous expression of AtMYB12-CaCHS resulted in the production of lower amounts of chlorogenic derivatives but similar levels of Ng glycosides compared with the CaCHS lines. Both CaCHS lines and the double transformant could be maintained over prolonged subcultures without Ng production losses, confirming long-term and stable CaCHS transgene expression.In no case did the inserted genes have a discernable effect on Sm production compared with the untransformed cultures, neither under control culture conditions or in cultures elicited with MeJA.

Transcriptomic profiling of sorghum leaves and roots responsive to drought stress at the seedling stage

Drought stress affects the growth and productivity of crop plants including sorghum. To study the molecular basis of drought tolerance in sorghum, we conducted the transcriptomic profiling of sorghum leaves and roots under drought stress using RNA-Seq method. A total of 510, 559, and 3 687 differentially expressed genes (DEGs) in leaves, 3 368, 5 093, and 4 635 DEGs in roots responding to mild drought, severe drought, and re-watering treatments were identified, respectively. Among them, 190 common DEGs in leaves and 1 644 common DEGs in roots were responsive to mild drought, severe drought, and re-watering environment. Gene Ontology (GO) enrichment analysis revealed that the GO categories related to drought tolerance include terms related to response to stimulus especially response to water deprivation, abscisic acid stimulus, and reactive oxygen species. The major transcription factor genes responsive to drought stress include heat stress transcription factor (HSF), ethylene-responsive transcription factor (ERF), Petunia NAM, Arabidopsis ATAF1/2 and CUC2 (NAC), WRKY transcription factor (WRKY), homeodomain leucine zipper transcription factor (HD-ZIP), basic helix-loop-helix transcription factor (bHLH), and V-myb myeloblastosis viral oncogene homolog transcription facotr (MYB). Functional protein genes for heat shock protein (HSPs), late-embryogenesis-abundant protein (LEAs), chaperones, aquaporins, and expansins might play important roles in sorghum drought tolerance. Moreover, the genomic regions enriched with HSP, expansin, and aquaporin genes responsive to drought stress could be used as powerful targets for improvement of drought tolerance in sorghum and other cereals. Overall, our results provide a genome-wide analysis of DEGs in sorghum leaves and roots under mild drought, severe drought, and re-watering environments. This study contributes to a better understanding of the molecular basis of drought tolerance of sorghum and can be useful for crop improvement.

MYB1 transcription factor regulation through floral scent in Cymbidium cultivar ‘Sael Bit’

Cymbidium belongs to Orchidaceae, one of the most abundant angiosperm families. Cymbidium cultivars have a high aesthetic value and commercially valued characteristics such as a pleasant scent and a variety of flower colors. Cymbidium cultivar ‘Sael Bit’, which was developed by National Institute of Horticultural & Herbal Science (NIHHS), is highly fragrant with small green and light yellow flowers. An analysis of ‘Sael Bit’ floral scent emissions by an electronic nose showed that the petal and initial flowering stage emitted a strong scent. In an headspace-solid phase micro extraction-gas chromatography-mass spectrometry (HS-SPME-GC-MS) ‘Sael Bit’ study, we identified alkenes and benzenoids, such as tridecane, benzene, and Z-2-octenal, as major volatile organic compounds. V-myb myeloblastosis viral oncogene homolog (MYB) transcription factors (TFs) play a positive role in floral scent regulation. However, little is known about the MYB transcriptional regulation of floral scent biosynthesis. Due to the importance of the plant’s biology, we need to study the floral scent MYB TFs in this plant. In the present work, we isolated and cloned the full-length cDNA of Cymbidium cultivar ‘Sael Bit’ MYB1 from fully open flower. CsMYB1 encodes an 840-bp open reading frame and 280 amino acids by functional characterization. In q-RT-PCR expression analysis, CsMYB1 TF showed the highest expression in petals and columns compared to sepals and labella. Moreover, this MYB gene expressed various flower developmental stages and was highly expressed at the fully open flower stage. The phenylpropanoid/benzenoid and ester compounds responsible genes expressed through CsMYB1 TF regulation in q-RT-PCR analysis. According to these results, we suggest that CsMYB1 might be involved in the regulation of phenylpropanoid/benzenoid genes in floral scent profile. In future studies, we will focus on develop regulation models of floral scent in Cymbidium cultivar ‘Sael Bit’.

Expression Patterns of MYB (V-myb Myeloblastosis Viral Oncogene Homolog) Gene Family in Resistant and Susceptible Tung Trees Responding to Fusarium Wilt Disease

Vernicia fordii (tung oil tree) is famous in the world for its production of tung oil. Unfortunately, it was infected by the soil-borne fungus Fusarium oxysporum f. sp. fordii 1 (Fof-1) and suffered serious wilt disease. Conversely, its sister species V. montana is highly resistant to Fof-1. The MYB (v-myb myeloblastosis viral oncogene homolog) transcription factors were activated during the pathogen Fof-1 infection according to our previous comparative transcriptomic results. Depending on whether the sequence has a complete MYB-DNA-binding domain, a total of 75 VfMYB and 77 VmMYB genes were identified in susceptible V. fordii and resistant V. montana, respectively. In addition, we detected 49 pairs of one-to-one orthologous Vf/VmMYB genes with the reciprocal-best BLAST-hits (RBH)method. In order to investigate the expression modes and the internal network of MYB transcription factors in the two species responding to Fusarium wilt disease, the expressions of Vf/VmMYBs were then investigated and we found that most orthologous Vf/VmMYB genes exhibited similar expression patterns during the Fof-1 infection. However, four pairs of Vf/VmMYB genes, annotated as unknown proteins and mediator of root architecture, demonstrated absolute opposite expression patterns in the two Vernicia species responding to Fof-1. The interaction network of VmMYB genes were further constructed using weighted gene co-expression network analysis (WGCNA) method and four hub genes showing extremely high interaction with the other 1157 genes were identified. RT-qPCR result verified the opposite expression pattern of the hub gene VmMYB011 and VmMYB041 in two Vernicia species. In summary, co-expression network of the Vf/VmMYBs and significantly opposite related pairs of genes in resistant and susceptible Vernicia species provided knowledge for understanding the molecular basis of Vernicia responding to Fusarium wilt disease.

Differential expression of six chicken genes associated with fatness traits in a divergently selected broiler population

A genome-wide association study has shown a number of chicken (Gallus gallus) single nucleotide polymorphism (SNP) markers to be significantly associated with abdominal fat content in Northeast Agricultural University (NEAU) broiler lines selected divergently for abdominal fat content (NEAUHLF). The six significant SNPs are located in the kinase insert domain receptor (KDR), tumor suppressor candidate 3 (TUSC3), phosphoribosyl pyrophosphate amidotransferase (PPAT), exocyst complex component 1 (EXOC1), v-myb myeloblastosis viral oncogene homolog (avian)-like 2 (MYBL2) and KIAA1211 (undefined) genes. In this study, the expression levels of these genes were investigated in both abdominal fat and liver tissues using 32 14th generation chickens from the NEAUHLF. The levels of expression of KDR in abdominal fat and KDR and TUSC3 in liver differed significantly between the two lines. The expression level of KDR in the abdominal fat was significantly correlated with the abdominal fat weight (AFW) and abdominal fat percentage (AFP). The expression levels of KDR, TUSC3 and PPAT in liver were significantly correlated with AFW and AFP, indicating that the six genes, especially KDR and TUSC3, could be associated with fat traits in domestic chickens. This study could provide insight into the mechanisms underlying the formation of abdominal fat in chickens.

Anthocyanin profile and gene expression in berry skin of two redVitis viniferagrape cultivars that are sunlight dependent versus sunlight independent

Background and Aims When sunlight was excluded (opacity coefficient 100%) from fruitset until maturity, the red grape cultivar Jingxiu failed to colour while Jingyan (Jingxiu × white grape Xiangfei) developed red colour. The aim of this study was to gain insight into the genetic background that may be responsible for sunlight-dependent versus sunlight-independent anthocyanin biosynthesis in berry skin. Methods and Results The composition and concentration of anthocyanins via high-performance liquid chromatography, and the transcriptional level of structural and regulatory genes via real-time polymerase chain reaction were investigated. Light exclusion suppressed all the anthocyanins in Jingxiu berry skin, but did not change the proportion of the various anthocyanins in Jingyan. UDP-glucose:flavonoid 3-O-glucosyltransferase (UFGT) and a v-myb myeloblastosis viral oncogene homolog transcription factor (VvMYBA1) were expressed at a high level in sunlight-exposed Jingxiu, sunlight-exposed and excluded Jingyan, but not in sunlight-excluded Jingxiu, which correlated with phenotypic colouration (red vs not red). There was no difference in the DNA sequence of the VvMYBA1b promoter region between Jingxiu and Jingyan. Conclusions The regulatory gene VvMYBA1 is involved in the regulation of anthocyanin biosynthesis via the structural gene UFGT, while there must be some other regulatory factors or post-transcriptional regulation mechanisms that differentially regulated VvMYBA1 in Jingxiu and Jingyan in response to sunlight. Significance of the Study The primary control mechanism of anthocyanin biosynthesis in the absence of sunlight would provide information for breeding sunlight-independent red grape cultivars, which would be valuable for cultivation in areas with low sunlight, e.g. glasshouses or other covered cropping.

Genome Wide DNA Methylation Analysis of Patients with Myelodysplastic Syndrome and Isolated Deletion (5q) Reveals Characteristic Methylation Profiles in Low and Intermediate-1 Risk Groups

AbstractAbstract 3801 BackgroundAberrant DNA methylation at promoter CpG islands is recognized as one of the hallmarks driving the pathogenesis of myeloid malignancies, especially myelodysplastic syndromes (MDS). Shen et al. recently characterized an algorithm of 10 aberrantly methylated gene loci which was predictive for overall survival and progression-free survival in a large cohort of MDS patients. A particular cytogenetic subgroup of MDS patients with a deletion on the long arm of chromosome 5 (5q-syndrome) has been shown to benefit from treatment with lenalidomide. However, the exact underlying molecular mechanism of MDS with isolated deletion (5q) is still not understood. To further elucidate on the role of deregulated DNA methylation we analyzed DNA-methylation profiles of bone marrow cells from patients with MDS with an isolated deletion (5q). MethodsAll patients were diagnosed and treated within a German multicenter trial investigating the safety of Lenalidomide in patients with low risk myelodysplastic syndromes and an isolated deletion (5q) after informed consent and according to the declaration of Helsinki. Bone marrow cells of 47 MDS patients with deletion (5q) at initial diagnosis were analyzed (median age 70 years, range 41 – 88 years, IPSS score: low n= 22; intermediate-1 n = 25).DNA was extracted using the QIAGEN Allprep Kit® (Qiagen, Hilden, Germany). Genome wide DNA methylation analysis was performed using the HumanMethylation450 BeadChip (Illumina, San Diego, USA). Differential methylation of CpGs was defined by a minimum mean methylation difference of 15% as expressed by the beta-value of the array data and statistical significance set at q ≤ 0.01 according to the Benjamini-Hochberg-method for multiple significance testing. Analysis of array data was performed using Genome-Studio Software® (Illumina, San Diego, USA), Qlucore Omics explorer 2.3 (Qlucore software. Lund, Sweden) and Microsoft Excel 10.1® (Microsoft Software, Redmond, USA). Gene ontology analysis was performed using GATHER (http://http://gather.genome.duke.edu/). ResultsUsing a q-value of ≤ 0.05 for the beta-value of the array and excluding gender-specific chromosomal CpGs, 473,929 CpGs were evaluable for analysis. Gene Ontology analysis using the GATHER Tool showed a significant enrichment of genes mapped to 5q31 (p<0.0001).Highly significant differential methylation profiles between MDS patients with isolated (5q) were found between patients with low and intermediate-1 IPSS score.CpGs differentially hypermethylated in intermediate-1 risk versus lows risk patients affected the coding regions of interesting candidate genes such as platelet-derived growth factor receptor, beta polypeptide (PDGFRB), clathrin interactor 1 (CLINT1), both located at 5q33 and suspected to be involved in the pathogenesis of 5q deleted MDS. Furthermore, transcriptional regulators such as proline, glutamate and leucine rich protein 1 (PELP1), v-myb myeloblastosis viral oncogene homolog (avian) (MYB), genes known to be involved in cancer like trichorhinophalangeal 1(TRPS1), and tumor suppressors like forkhead box P1 (FOXP1) and genes thought to be involved in the pathogenesis of MDS like Minichromosome maintenance protein2(MCM2) did show differentially DNA-methylation according to our selection criteria. ConclusionsWe present a comprehensive genome wide methylation analysis of MDS patients with an isolated deletion (5q) with low and intermediate-1 risk according to IPSS. Thereby we detected sets of significantly differentially methylated CpGs between both risk groups. Correlation of these data to clinical parameters might help to further elucidate the contribution of aberrant methylation to the phenotype of MDS with isolated deletion (5q) and could possibly help establishing novel prognostic markers based on differential methylation. Moreover, unraveling the role of aberrant methylation patterns might result in new therapeutic treatment approaches at least in a subset of patients. Disclosures:Nowak:Celgene: Research Funding. Platzbecker:Celgene: Honoraria, Membership on an entity’s Board of Directors or advisory committees, Research Funding. Giagounidis:Celgene: Honoraria, Membership on an entity’s Board of Directors or advisory committees. Götze:Celgene: Honoraria, Membership on an entity’s Board of Directors or advisory committees. Ottmann:Celgene: Clinical trial participation Other. Haase:Celgene: Honoraria, Membership on an entity’s Board of Directors or advisory committees. Schlenk:Celgene: Research Funding. Ganser:Celgene: Research Funding. Germing:Celgene: Honoraria, Membership on an entity’s Board of Directors or advisory committees, Research Funding. Hofmann:Celgene: Honoraria, Research Funding. Nolte:Celgene: Research Funding.

Transcription Regulation Network Analysis of MCF7 Breast Cancer Cells Exposed to Estradiol

In breast cancer, estrogen receptors have been demonstrated to interact with transcription factors to regulate target gene expression. However, high-throughput identification of the transcription regulation relationship between transcription factors and their target genes in response to estradiol is still in its infancy. Thus, the objective of our study was to interpret the transcription regulation network of MCF7 breast cancer cells exposed to estradiol. In this work, GSE11352 microarray data were used to identify differentially expressed genes (DEGs). Our results showed that the MYB (v-myb myeloblastosis viral oncogene homolog [avian]), PGR (progesterone receptor), and MYC (v-myc myelocytomatosis viral oncogene homolog [avian]) were hub nodes in our transcriptome network, which may interact with ER and, in turn, regulate target gene expression. MYB can up-regulate MCM3 (minichromosome maintenance 3) and MCM7 expression; PGR can suppress BCL2 (B-cell lymphoma 2) expression; MYC can inhibit TGFB2 (transforming growth factor, beta 2) expression. These genes are associated with breast cancer progression via cell cycling and the TGFβ signaling pathway. Analysis of transcriptional regulation may provide a better understanding of molecular mechanisms and clues to potential therapeutic targets in the treatment of breast cancer.

MYBL2 (v-myb myeloblastosis viral oncogene homolog (avian)-like 2)

Review on MYBL2 (v-myb myeloblastosis viral oncogene homolog (avian)-like 2), with data on DNA, on the protein encoded, and where the gene is implicated.

MYBL1 (v-myb myeloblastosis viral oncogene homolog (avian)-like 1)

Review on MYBL1 (v-myb myeloblastosis viral oncogene homolog (avian)-like 1), with data on DNA, on the protein encoded, and where the gene is implicated.

MYB (v-myb myeloblastosis viral oncogene homolog (avian))

Review on MYB (v-myb myeloblastosis viral oncogene homolog (avian)), with data on DNA, on the protein encoded, and where the gene is implicated.

MYB transcriptionally regulates the miR-155 host gene in chronic lymphocytic leukemia

AbstractElevated levels of microRNA miR-155 represent a candidate pathogenic factor in chronic B-lymphocytic leukemia (B-CLL). In this study, we present evidence that MYB (v-myb myeloblastosis viral oncogene homolog) is overexpressed in a subset of B-CLL patients. MYB physically associates with the promoter of miR-155 host gene (MIR155HG, also known as BIC, B-cell integration cluster) and stimulates its transcription. This coincides with the hypermethylated histone H3K4 residue and spread hyperacetylation of H3K9 at MIR155HG promoter. Our data provide evidence of oncogenic activities of MYB in B-CLL that include its stimulatory role in MIR155HG transcription.

Gene expression profiles of primary HPV16- and HPV18-infected early stage cervical cancers and normal cervical epithelium: identification of novel candidate molecular markers for cervical cancer diagnosis and therapy

With the goal of identifying genes with a differential pattern of expression between invasive cervical carcinomas (CVX) and normal cervical keratinocytes (NCK), we used oligonucleotide microarrays to interrogate the expression of 14,500 known genes in 11 primary HPV16 and HPV18-infected stage IB–IIA cervical cancers and four primary normal cervical keratinocyte cultures. Hierarchical cluster analysis of gene expression data identified 240 and 265 genes that exhibited greater than twofold up-regulation and down-regulation, respectively, in primary CVX when compared to NCK. Cyclin-dependent kinase inhibitor 2A (CDKN2A/p16), mesoderm-specific transcript, forkhead box M1, v-myb myeloblastosis viral oncogene homolog (avian)-like2 (v-Myb), minichromosome maintenance proteins 2, 4, and 5, cyclin B1, prostaglandin E synthase (PTGES), topoisomerase II alpha (TOP2A), ubiquitin-conjugating enzyme E2C, CD97 antigen, E2F transcription factor 1, and dUTP pyrophosphatase were among the most highly overexpressed genes in CVX when compared to NCK. Down-regulated genes in CVX included transforming growth factor beta 1, transforming growth factor alpha, CFLAR, serine proteinase inhibitors (SERPING1 and SERPINF1), cadherin 13, protease inhibitor 3, keratin 16, and tissue factor pathway inhibitor-2 (TFPI-2). Differential expression of some of these genes including CDKN2A/p16, v-Myb, PTGES, and TOP2A was validated by quantitative real-time PCR. Flow cytometry on primary CVX and NCK and immunohistochemical staining of formalin fixed paraffin-embedded tumor specimens from which primary CVX cultures were derived as well as from a separate set of invasive cervical cancers confirmed differential expression of the CDKN2A/p16 and PTGES markers on CVX versus NCK. These results identify several genes that are coordinately disregulated in cervical cancer, likely representing common signaling pathways triggered by HPV transformation. Moreover, these data obtained with highly purified primary tumor cultures highlight novel molecular features of human cervical cancer and provide a foundation for the development of new type-specific diagnostic and therapeutic strategies for this disease.

Profiling molecular targets of TGF-β1 in prostate fibroblast-to-myofibroblast transdifferentiation

The development of age-related proliferative disorders of the prostate gland is supported by transdifferentiation and cellular senescence processes in the stroma. Both processes are involved in remodeling of stromal tissue, as observed in benign prostatic hyperplasia (BPH), and in “reactive stroma” adjacent to prostate cancer (PCa). It has been assumed that TGF-β1 plays a key role in the aging prostate by inducing premature senescence and favoring myofibroblast differentiation. Therefore, we evaluated the stromal cell phenotypes of human primary adult prostatic fibroblasts ( n = 3) and the molecular and cellular mechanisms of growth arrest after treatment with TGF-β1 and of in vitro cellular senescence. Microarray analysis, quantitative PCR, immunofluorescence and western blot revealed that cellular senescence and transdifferentiation of fibroblasts have distinct underlying mechanisms, pathways and gene and protein expression profiles in human PrSCs. In clear contrast to senescent cells, TGF-β1-treated cells morphologically transdifferentiated into myofibroblasts with dense cytoskeletal fibers and increased expression of smooth muscle cell α-actin, calponin and tenascin. TGF-β1 induced neither expression of senescence-associated markers nor genes involved in terminal growth arrest, such as senescence-associated beta-galactosidase and cyclin-dependent kinase (cdk) inhibitors p16 Ink4A and p21 Cip1 but increased p15 Ink4B protein expression. Differentiation inhibitor (Id-1) protein level down-regulation was observed under both conditions. Genes specifically up-regulated by transdifferentiation but not by cellular senescence of PrSCs were metalloproteinase 1 tissue inhibitor ( Timp1), transgelin ( Tagln), gamma 2 actin ( Actg2), plasminogen activator inhibitor 1 ( Serpinel), insulin-like growth factor binding protein 3 ( Igfbp3), parathyroid hormone-like hormone ( Pthlp), Tgfb-1, four and a half LIM domains 2 ( Fhl-2), hydrogen peroxide-inducible clone 5 ( Hic5) and cartilage oligomeric matrix protein (Comp). Other genes, such as Cdc28 protein kinase 1 ( Cks1b), v-myb myeloblastosis viral oncogene homolog ( MybL2), pyruvate kinase, muscle 2 ( Pkm2) and Forkhead box M1 ( FoxM1), were down-regulated only upon TGF-β1 treatment but not by cellular senescence. Pyruvate dehydrogenase kinase 3 ( Pdk3) and connective tissue growth factor ( Ctgf) were up-regulated and hyaluronan synthase 3 ( Has3) down-regulated under both conditions. Moreover, GageC1, a prostate/testis-specific protein overexpressed in symptomatic BPH and PCa was induced in transdifferentiated stromal cells. Genes such as GageC1 could be promising targets for therapeutic inhibitors of stromal tissue remodeling and progression of BPH and PCa.