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1.
Carcinogenesis ; 36(6): 622-31, 2015 Jun.
Article in English | MEDLINE | ID: mdl-25827435

ABSTRACT

SWI/SNF chromatin remodeling complexes constitute a highly related family of multi-subunit complexes to modulate transcription, and SWI/SNF subunit genes are collectively mutated in 20% of all human cancers. Bcl11b is a SWI/SNF subunit and acts as a haploinsufficient tumor suppressor in leukemia/lymphomas. Here, we show expression of Bcl11b in intestinal crypt cells and promotion of intestinal tumorigenesis by Bcl11b attenuation in Apc (min/+) mice. Of importance, mutations or allelic loss of BCL11B was detected in one-third of human colon cancers. We also show that attenuated Bcl11b activity in the crypt base columnar (CBC) cells expressing the Lgr5 stem cell marker enhanced regeneration of intestinal epithelial cells after the radiation-induced injury. Interestingly, BCL11B introduction in human cell lines downregulated transcription of ß-catenin target genes, whereas Bcl11b attenuation in Lgr5(+) CBCs increased expression of ß-catenin targets including c-Myc and cyclin D1. Together, our results argue that Bcl11b impairment promotes tumor development in mouse and human intestine at least in part through deregulation of ß-catenin pathway.


Subject(s)
Cell Transformation, Neoplastic/genetics , Chromosomal Proteins, Non-Histone/genetics , Colonic Neoplasms/genetics , Repressor Proteins/genetics , Transcription Factors/genetics , Tumor Suppressor Proteins/genetics , beta Catenin/metabolism , Adenoma/classification , Adenoma/genetics , Animals , Caco-2 Cells , Cell Line, Tumor , Cell Proliferation , Colonic Neoplasms/classification , Cyclin D1/biosynthesis , HCT116 Cells , HEK293 Cells , Humans , Intestinal Mucosa/metabolism , Intestinal Mucosa/radiation effects , Mice , Mice, Inbred C57BL , Mice, Knockout , Polymorphism, Single Nucleotide , Proto-Oncogene Proteins c-myc/biosynthesis , Receptors, G-Protein-Coupled/biosynthesis , Repressor Proteins/biosynthesis , Tumor Suppressor Proteins/biosynthesis , Wnt Proteins/metabolism , Wnt Signaling Pathway , beta Catenin/biosynthesis , beta Catenin/genetics
2.
Cancer Sci ; 106(4): 461-5, 2015 Apr.
Article in English | MEDLINE | ID: mdl-25613934

ABSTRACT

Human T-cell leukemia virus type 1 (HTLV-1) is the etiological agent of adult T cell leukemia (ATL), which is an aggressive form of T-cell malignancy. HTLV-1 oncoproteins, Tax and HBZ, play crucial roles in the immortalization of T-cells and/or leukemogenesis by dysregulating the cellular functions in the host. Recent studies show that HTLV-1-infected T-cells have reduced expression of the BCL11B tumor suppressor protein. In the present study, we explored whether Tax and/or HBZ play a role in downregulating BCL11B in HTLV-1-infected T-cells. Lentiviral transduction of Tax in a human T-cell line repressed the expression of BCL11B at both the protein and mRNA levels, whereas the transduction of HBZ had little effect on the expression. Tax mutants with a decreased activity for the NF-κB, CREB or PDZ protein pathways still showed a reduced expression of the BCL11B protein, thereby implicating a different function of Tax in BCL11B downregulation. In addition, the HTLV-2 Tax2 protein reduced the BCL11B protein expression in T-cells. Seven HTLV-1-infected T-cell lines, including three ATL-derived cell lines, showed reduced BCL11B mRNA and protein expression relative to an uninfected T-cell line, and the greatest reductions were in the cells expressing Tax. Collectively, these results indicate that Tax is responsible for suppressing BCL11B protein expression in HTLV-1-infected T-cells; Tax-mediated repression of BCL11B is another mechanism that Tax uses to promote oncogenesis of HTLV-1-infected T-cells.


Subject(s)
Gene Products, tax/metabolism , Human T-lymphotropic virus 1/pathogenicity , Leukemia-Lymphoma, Adult T-Cell/virology , Repressor Proteins/biosynthesis , Tumor Suppressor Proteins/biosynthesis , Basic-Leucine Zipper Transcription Factors/genetics , Basic-Leucine Zipper Transcription Factors/metabolism , Cell Line, Transformed , Cyclic AMP Response Element-Binding Protein/metabolism , Down-Regulation , Gene Products, tax/genetics , HEK293 Cells , Human T-lymphotropic virus 1/genetics , Human T-lymphotropic virus 2/genetics , Human T-lymphotropic virus 2/pathogenicity , Humans , Jurkat Cells , NF-kappa B/metabolism , Retroviridae Proteins , T-Lymphocytes/virology , Viral Proteins/genetics , Viral Proteins/metabolism
3.
Int Immunol ; 27(4): 205-15, 2015 Apr.
Article in English | MEDLINE | ID: mdl-25422283

ABSTRACT

If Bcl11b activity is compromised, CD4(+)CD8(+) double-positive (DP) thymocytes produce a greatly increased fraction of innate CD8(+) single-positive (SP) cells highly producing IFN-γ, which are also increased in mice deficient of genes such as Itk, Id3 and NF-κB1 that affect TCR signaling. Of interest, the increase in the former two is due to the bystander effect of IL-4 that is secreted by promyelocytic leukemia zinc finger-expressing NKT and γδT cells whereas the increase in the latter is cell intrinsic. Bcl11b zinc-finger proteins play key roles in T cell development and T cell-mediated immune response likely through TCR signaling. We examined thymocytes at and after the DP stage in Bcl11b (F/S826G) CD4cre, Bcl11b (F/+) CD4cre and Bcl11b (+/S826G) mice, carrying the allele that substituted serine for glycine at the position of 826. Here we show that Bcl11b impairment leads to an increase in the population of TCRαß(high)CD44(high)CD122(high) innate CD8SP thymocytes, together with two different developmental abnormalities: impaired positive and negative selection accompanying a reduction in the number of CD8SP cells, and developmental arrest of NKT cells at multiple steps. The innate CD8SP thymocytes express Eomes and secrete IFN-γ after stimulation with PMA and ionomycin, and in this case their increase is not due to a bystander effect of IL-4 but cell intrinsic. Those results indicate that Bcl11b regulates development of different thymocyte subsets at multiple stages and prevents an excess of innate CD8SP thymocytes.


Subject(s)
CD8-Positive T-Lymphocytes/cytology , CD8-Positive T-Lymphocytes/immunology , Cell Differentiation/immunology , Repressor Proteins/genetics , Tumor Suppressor Proteins/genetics , Animals , CD4-Positive T-Lymphocytes/immunology , Cells, Cultured , Hyaluronan Receptors/metabolism , Inhibitor of Differentiation Proteins/genetics , Interferon-gamma/biosynthesis , Interleukin-2 Receptor beta Subunit/metabolism , Interleukin-4/metabolism , Mice , Mice, Inbred C57BL , Mice, Transgenic , NF-kappa B/genetics , Natural Killer T-Cells/immunology , Protein-Tyrosine Kinases/genetics , Receptors, Antigen, T-Cell, alpha-beta/biosynthesis , Receptors, Antigen, T-Cell, alpha-beta/immunology , Receptors, Antigen, T-Cell, gamma-delta/biosynthesis , Receptors, Antigen, T-Cell, gamma-delta/immunology , Signal Transduction/immunology
4.
BMC Gastroenterol ; 14: 160, 2014 Sep 13.
Article in English | MEDLINE | ID: mdl-25218883

ABSTRACT

BACKGROUND: Intrahepatic cholestasis of pregnancy (ICP) is a cholestasis condition caused by elevated levels of serum bile acids that mainly occurs in the third trimester of pregnancy. Maternal symptoms include pruritus; elevation of transaminases, biliary enzymes, and bilirubin levels; and abnormal liver function tests. Fetal symptoms include spontaneous preterm labor, fetal distress, and intrauterine death. It is more prevalent in the Caucasians and is rarely found in Asian countries, including Japan. The etiology of ICP has been reported as involving various factors such as, environmental factors, hormone balance, and genetic components. The genetic factors include single-nucleotide polymorphisms (SNPs) in the genes of canalicular transporters, including ABCB4 and ABCB11. It has also been reported that the combination of these SNPs induces severe cholestasis and liver dysfunction. CASE PRESENTATION: Here, we report for the first time a 24-year Japanese case of severe ICP diagnosed by typical symptoms, serum biochemical analysis, and treated with the administration of ursodeoxycholic acid which improved cholestasis and liver injury and prevented fetal death. The sequence analysis showed SNPs reported their association with ICP in the ABCB11 (rs2287622, V444A) and ABCB4 (rs1202283, N168N) loci. CONCLUSION: The risk of ICP has been reported to be population-specific, and it is rare in the Japanese population. Our case was successfully treated with ursodeoxycholic acid and the genetic sequence analysis has supported the diagnosis. Because genetic variation in ABCB4 and ABCB11 has also been reported in the Japanese population, we need to be aware of potential ICP cases in pregnant Japanese women although further studies are necessary.


Subject(s)
Cholagogues and Choleretics/therapeutic use , Cholestasis, Intrahepatic/drug therapy , Pregnancy Complications/drug therapy , Ursodeoxycholic Acid/therapeutic use , Asian People , Female , Humans , Pregnancy , Treatment Outcome , Young Adult
5.
Mech Dev ; 130(9-10): 482-92, 2013.
Article in English | MEDLINE | ID: mdl-23727454

ABSTRACT

Rodent incisors maintain the ability to grow continuously and their labial dentin is covered with enamel. Bcl11b zinc-finger transcription factor is expressed in ameloblast progenitors in mouse incisors and its absence in Bcl11b(KO/KO) mice results in a defect in embryonic tooth development. However, the role of Bcl11b in incisor maintenance in adult tissue was not studied because of death at birth in Bcl11b(KO/KO) mice. Here, we examined compound heterozygous Bcl11b(S826G/KO) mice, one allele of which has an amino acid substitution of serine at position 826 for glycine, that exhibited hypoplastic maxillary incisors with lower concentrations of minerals at the enamel and the dentin, accompanying the maxillary bone hypoplasia. Histological examinations revealed hypoplasia of the labial cervical loop in incisors, shortening of the ameloblast progenitor region, and impairment in differentiation and proliferation of ameloblast-lineage cells. Interestingly, however, juvenile mice at 5days after birth did not show marked change in these phenotypes. These results suggest that attenuated Bcl11b activity impairs ameloblast progenitors and incisor maintenance. The number of BrdU label-retaining cells, putative stem cells, was lower in Bcl11b(S826G/KO) incisors, which suggests the incisor hypoplasia may be in part a result of the decreased number of stem cells. Interestingly, the level of Shh and FGF3 expressions, which are assumed to play key roles in the development and maintenance of ameloblasts and odontoblasts, was not decreased, though the expressed areas were more restricted in ameloblast progenitor and mesenchyme regions of Bcl11b(S826G/KO) incisors, respectively. Those data suggest that the incisor maintenance by Bcl11b is not directly related to the FGF epithelial-mesenchymal signaling loop including Shh but is intrinsic to ameloblast progenitors and possibly stem cells.


Subject(s)
Ameloblasts/metabolism , Gene Expression Regulation, Developmental , Incisor/metabolism , Maxilla/metabolism , Repressor Proteins/genetics , Stem Cells/metabolism , Tumor Suppressor Proteins/genetics , Age Factors , Ameloblasts/cytology , Amino Acid Substitution , Animals , Animals, Newborn , Cell Count , Cell Differentiation , Fibroblast Growth Factor 3/genetics , Fibroblast Growth Factor 3/metabolism , Hedgehog Proteins/genetics , Hedgehog Proteins/metabolism , Heterozygote , Incisor/cytology , Incisor/growth & development , Male , Maxilla/cytology , Maxilla/growth & development , Mice , Mice, Knockout , Repressor Proteins/deficiency , Signal Transduction , Stem Cells/cytology , Transcription, Genetic , Tumor Suppressor Proteins/deficiency
6.
Cancer Sci ; 104(8): 1009-16, 2013 Aug.
Article in English | MEDLINE | ID: mdl-23663453

ABSTRACT

Bcl11b is a haploinsufficient tumor suppressor, mutations or deletion of which has been found in 10-16% of T-cell acute lymphoblastic leukemias. Bcl11b(KO) (/+) heterozygous mice are susceptible to thymic lymphomas, a model of T-cell acute lymphoblastic leukemia, when γ-irradiated, and irradiated Bcl11b(KO) (/+) mice generate clonally expanding or premalignant thymocytes before thymic lymphoma development. Cells with radiation-induced DNA damages are assumed to be the cells of origin in tumors; however, which thymocyte is the tumor cell origin remains obscure. In this study we generated Bcl11b(flox/+) ;Lck-Cre and Bcl11b(flox/+) ;CD4-Cre mice; in the former, loss of one Bcl11b allele occurs in thymocytes at the immature CD4(-) CD8(-) stage, whereas in the latter the loss occurs in the more differentiated CD4(+) CD8(+) double-positive stage. We examined clonal expansion and differentiation of thymocytes in mice 60 days after 3 Gy γ-irradiation. Half (9/18) of the thymuses in the Bcl11b(flox/+) ;Lck-Cre group showed limited rearrangement sites at the T-cell receptor-ß (TCRß) locus, indicating clonal cell expansion, but none in the Bcl11b(flox/+) ;CD4-Cre group did. This indicates that the origin of the premalignant thymocytes is not in double-positive cells but immature thymocytes. Interestingly, those premalignant thymocytes underwent rearrangement at various different sites of the TCRα locus and the majority showed a higher expression of TCRß and CD8, and more differentiated phenotypes. This suggests the existence of a subpopulation of immature cells within the premalignant cells that is capable of proliferating and continuously producing differentiated thymocytes.


Subject(s)
Alleles , Neoplasms, Radiation-Induced/genetics , Precancerous Conditions/genetics , Repressor Proteins/genetics , Thymocytes/pathology , Thymocytes/radiation effects , Thymus Neoplasms/genetics , Tumor Suppressor Proteins/genetics , Animals , CD4-Positive T-Lymphocytes/metabolism , CD4-Positive T-Lymphocytes/pathology , CD4-Positive T-Lymphocytes/radiation effects , CD8-Positive T-Lymphocytes/metabolism , CD8-Positive T-Lymphocytes/pathology , CD8-Positive T-Lymphocytes/radiation effects , Cell Cycle/genetics , Cell Cycle/radiation effects , Cell Differentiation/genetics , Cell Differentiation/radiation effects , Cell Proliferation/radiation effects , Gamma Rays/adverse effects , Mice , Mice, Inbred C57BL , Neoplasms, Radiation-Induced/metabolism , Neoplasms, Radiation-Induced/pathology , Precancerous Conditions/metabolism , Precancerous Conditions/pathology , Receptors, Antigen, T-Cell, alpha-beta/genetics , Receptors, Antigen, T-Cell, alpha-beta/metabolism , Thymocytes/metabolism , Thymus Neoplasms/metabolism , Thymus Neoplasms/pathology
7.
Leuk Res ; 36(8): 1035-40, 2012 Aug.
Article in English | MEDLINE | ID: mdl-22640496

ABSTRACT

Genetic changes in T-ALL are classified into type A abnormalities leading to arrest at a specific stage of T-cell differentiation and type B abnormalities that target cellular processes including cell cycle regulation. Mutations and deletion of a BCL11B haploinsuffiecient tumor suppressor allele have been found in 10-16% of T-ALL subgroups. Analysis of Bcl11b(KO/+) mice revealed impaired T-cell differentiation at two different stages and attenuation of γ-ray induced cell-cycle arrest at S/G2/M phase in immature CD8 single positive cells. Hence, those phenotypes provided by loss of a Bcl11b allele favor that Bcl11b mutation belongs to type B abnormalities.


Subject(s)
Cell Cycle/genetics , Cell Differentiation/genetics , Leukemia/genetics , Repressor Proteins/genetics , Thymocytes/physiology , Tumor Suppressor Proteins/genetics , Alleles , Animals , Cell Transformation, Neoplastic/genetics , Cell Transformation, Neoplastic/pathology , Gene Deletion , Leukemia/metabolism , Leukemia/pathology , Loss of Heterozygosity/physiology , Mice , Mice, Inbred BALB C , Mice, Knockout , Phenotype , Repressor Proteins/metabolism , Repressor Proteins/physiology , Thymocytes/metabolism , Tumor Suppressor Proteins/metabolism , Tumor Suppressor Proteins/physiology
8.
Cell Signal ; 24(5): 1047-52, 2012 May.
Article in English | MEDLINE | ID: mdl-22245141

ABSTRACT

BCL11B is a C2H2 zinc finger transcription factor that acts as a haploinsufficient tumor suppressor. Mutations and deletion in the human orthologue BCL11B have been identified in human T-cell acute lymphoblastic leukemia (T-ALL) and a mouse model of thymic lymphomas. Bcl11b(KO/+)p53(KO/+) doubly heterozygous mice, but not Bcl11b(KO/+) heterozygous mice, spontaneously develop thymic lymphomas at a high frequency, suggesting cooperativity of BCL11B and p53 in cancer development. In this study, we have examined whether or not BCL11B directly affects the p53 signaling pathway including HDM2, a ubiquitin ligase for p53 degradation. The p53 pathway regulates cell proliferation and the response to DNA damages to maintain genome integrity. Here we show that BCL11B binds to human HDM2-P2 promoter by ChIP (chromatin immuno-precipitation) assay and inhibits HDM2 expression in a p53-dependent manner. Deletion of the distal p53 responsive element in HDM2 promoter region or the lack of p53 in HCT116 cells greatly reduced the repressive effect of BCL11B on HDM2-P2 promoter activity. The repressive activity was alleviated in γ-ray induced DNA damage conditions that activate p53, suggesting interaction between BCL11B and p53 for HDM2 expression. These date suggest that BCL11B affects the activity of the p53-HDM2 feedback loop in basal and irradiated conditions. This may be a mechanism underlying the leukemic transformation in T-ALL and in Bcl11b(KO/+)p53(KO/+) mouse thymocytes.


Subject(s)
Gene Expression Regulation , Proto-Oncogene Proteins c-mdm2/metabolism , Repressor Proteins/physiology , Tumor Suppressor Protein p53/metabolism , Tumor Suppressor Proteins/physiology , Cell Line , DNA Damage , Gamma Rays , Genes, Reporter , Humans , Luciferases, Renilla/biosynthesis , Luciferases, Renilla/genetics , Promoter Regions, Genetic , Protein Binding , Proto-Oncogene Proteins c-mdm2/genetics , Repressor Proteins/metabolism , Transcription, Genetic , Tumor Suppressor Proteins/metabolism
9.
Exp Anim ; 60(4): 355-61, 2011.
Article in English | MEDLINE | ID: mdl-21791875

ABSTRACT

BCL11B/CTIP2 zinc-finger transcription factor is expressed in various types of cells in many different tissues. This study showed that BCL11B is expressed in the nucleus of the outer hair cells of the mouse cochlea, degeneration of which is known to cause deafness and presbycusis or age-related hearing loss (AHL). We tested whether or not Bcl11b heterozygosity would affect AHL in mice. Analysis of auditory brainstem responses revealed AHL in Bcl11b (+/-) heterozygous, but not wild-type, mice, which was evident as early as 3 months after birth. Histological abnormalities were observed in the outer hair cells of the Bcl11b (+/-) mice at 6 months of age with hearing loss. These results suggest that the AHL observed in Bcl11b (+/-) mice is the result of impairment of the outer hair cells and that BCL11B activity is required for the maintenance of outer hair cells and normal hearing.


Subject(s)
Hair Cells, Auditory/pathology , Presbycusis/genetics , Presbycusis/pathology , Repressor Proteins/genetics , Tumor Suppressor Proteins/genetics , Animals , Cell Nucleus/genetics , Disease Models, Animal , Evoked Potentials, Auditory, Brain Stem , Hair Cells, Auditory/metabolism , Mice , Mice, Inbred BALB C , Polymorphism, Genetic , Repressor Proteins/metabolism , Tumor Suppressor Proteins/metabolism
10.
Int J Radiat Oncol Biol Phys ; 77(1): 235-43, 2010 May 01.
Article in English | MEDLINE | ID: mdl-20394855

ABSTRACT

PURPOSE: To characterize, in the setting of gamma-ray-induced atrophic thymus, probable prelymphoma cells showing clonal growth and changes in signaling, including DNA damage checkpoint. METHODS AND MATERIALS: A total of 111 and 45 mouse atrophic thymuses at 40 and 80 days, respectively, after gamma-irradiation were analyzed with polymerase chain reaction for D-J rearrangements at the TCRbeta locus, flow cytometry for cell cycle, and Western blotting for the activation of DNA damage checkpoints. RESULTS: Limited D-J rearrangement patterns distinct from normal thymus were detected at high frequencies (43 of 111 for 40-day thymus and 21 of 45 for 80-day thymus). Those clonally expanded thymocytes mostly consisted of CD4(+)CD8(+) double-positive cells, indicating the retention of lineage capability. They exhibited pausing at a late G1 phase of cell cycle progression but did not show the activation of DNA damage checkpoints such as gammaH2AX, Chk1/2, or p53. Of interest is that 17 of the 52 thymuses showing normal D-J rearrangement patterns at 40 days after irradiation showed allelic loss at the Bcl11b tumor suppressor locus, also indicating clonal expansion. CONCLUSION: The thymocytes of clonal growth detected resemble human chronic myeloid leukemia in possessing self-renewal and lineage capability, and therefore they can be a candidate of the lymphoma-initiating cells.


Subject(s)
DNA Damage , Lymphoma/pathology , Neoplasms, Radiation-Induced/pathology , Precancerous Conditions/pathology , T-Lymphocytes/radiation effects , Thymus Gland/radiation effects , Adaptor Proteins, Signal Transducing , Animals , Atrophy/pathology , CD4-Positive T-Lymphocytes/pathology , CD4-Positive T-Lymphocytes/radiation effects , CD8-Positive T-Lymphocytes/pathology , CD8-Positive T-Lymphocytes/radiation effects , Cell Cycle/physiology , Cell Cycle Proteins , Cell Proliferation/radiation effects , Gamma Rays , Gene Deletion , Gene Rearrangement, T-Lymphocyte , Histones/metabolism , Lymphoma/genetics , Lymphoma/metabolism , Mice , Neoplasms, Radiation-Induced/genetics , Neuropilin-1/metabolism , Nuclear Proteins/metabolism , Precancerous Conditions/genetics , Precancerous Conditions/metabolism , T-Lymphocytes/metabolism , T-Lymphocytes/pathology , Thymus Gland/pathology , Trans-Activators/metabolism , Tumor Suppressor Protein p53/metabolism
11.
Cancer Sci ; 101(6): 1347-53, 2010 Jun.
Article in English | MEDLINE | ID: mdl-20384631

ABSTRACT

Bcl11b encodes a zinc-finger transcription factor and functions as a haploinsufficient tumor suppressor gene. Bcl11b(KO/KO) mice exhibit differentiation arrest of thymocytes during beta-selection as has been observed with other mouse models involving knockouts of genes in the Wnt/beta-catenin signaling pathway. Recurrent chromosomal rearrangement at the BCL11B locus occurs in human T-cell leukemias, but it is not clear how such rearrangement would contribute to lymphomagenesis. To address this issue, we studied clonal cell growth, cell number, and differentiation of thymocytes in Bcl11b(KO/+) mice at different time points following gamma-irradiation. Analysis of D-J rearrangement at the T cell receptor beta-chain (TCRbeta) locus and cell surface markers by flow cytometry revealed two distinct populations of clonally growing thymocytes. In one population, thymocytes share a common D-J rearrangement but retain the capacity to differentiate. In contrast, thymocytes in the second population have lost their ability to differentiate. Since the capacity to self renew and differentiate into multiple cell lineages are fundamental properties of adult stem cells, the differentiation competent population of thymocytes that we have isolated could potentially function as cancer stem cells. We also demonstrate increased expression of beta-catenin, a well-known oncogenic protein, in Bcl11b(KO/+) thymocytes. Collectively, the Bcl11b(KO/+) genotype contributes to clonal expansion and differentiation arrest in part through an increase in the level of beta-catenin.


Subject(s)
Lymphoma/etiology , Repressor Proteins/physiology , T-Lymphocytes/radiation effects , Tumor Suppressor Proteins/physiology , Animals , Cell Cycle/radiation effects , Cell Differentiation/radiation effects , Cell Size/radiation effects , Gamma Rays , Heterozygote , Mice , Mice, Inbred BALB C , Receptors, Antigen, T-Cell/physiology , Receptors, Interleukin-7/analysis , Repressor Proteins/genetics , Signal Transduction , T-Lymphocytes/pathology , Thymus Neoplasms/etiology , Tumor Suppressor Proteins/genetics , beta Catenin/analysis , beta Catenin/physiology
12.
Biochem Biophys Res Commun ; 373(2): 282-5, 2008 Aug 22.
Article in English | MEDLINE | ID: mdl-18558082

ABSTRACT

Bcl11b is a haploinsufficient tumor suppressor gene and expressed in many tissues such as thymus, brain and skin. Irradiated Bcl11b+/- heterozygous mice mostly develop thymic lymphomas, but the preference of Bcl11b inactivation for thymic lymphomas remains to be addressed. We produced Bcl11b+/- heterozygous and Bcl11b wild-type mice of p53+/- background and compared their incidence of gamma-ray induced thymic lymphomas. Majority of the tumors in p53+/- mice were skin tumors, and only 5 (36%) of the 14 tumors were thymic lymphomas. In contrast, Bcl11b+/-p53+/- doubly heterozygous mice developed thymic lymphomas at the frequency of 27 (79%) of the 34 tumors developed (P=0.008). This indicates the preference of Bcl11b impairment for thymic lymphoma development. We also analyzed loss of the wild-type alleles in the 27 lymphomas, a predicted consequence given by gamma-irradiation. However, the loss frequency was low, only six (22%) for Bcl11b and five (19%) for p53. The frequencies did not differ from those of spontaneously developed thymic lymphomas in the doubly heterozygous mice, though the latency of lymphoma development markedly differed between them. This suggests that the main contribution of irradiation at least in those mice is not for the tumor initiation by inducing allelic losses but probably for the promotion of thymic lymphoma development.


Subject(s)
DNA-Binding Proteins/genetics , Genetic Predisposition to Disease , Lymphoma/genetics , Neoplasms, Radiation-Induced/genetics , Repressor Proteins/genetics , Thymus Neoplasms/genetics , Tumor Suppressor Proteins/genetics , Animals , Gamma Rays , Genotype , Loss of Heterozygosity , Mice , Mice, Knockout , PTEN Phosphohydrolase/genetics , Tumor Suppressor Protein p53/genetics
13.
Biochem Biophys Res Commun ; 355(2): 538-42, 2007 Apr 06.
Article in English | MEDLINE | ID: mdl-17306224

ABSTRACT

Recurrent chromosomal rearrangements at BCL11B are found in human hematopoietic malignancies mostly of T-cell origin. However, it is unclear how this disruption contributes to oncogenesis, because the majority of leukemias express BCL11B from an undisrupted allele. Here, we show that Bcl11b(+/-)p53(+/-) mice exhibited greater susceptibility to lymphomas than Bcl11b(+/+)p53(+/-) mice but most lymphomas retained and expressed the wild-type Bcl11b allele. This strongly suggests that Bcl11b is haploinsufficient for suppression of thymic lymphoma development in mice of the p53(+/-) background, a situation in which functional loss of only one allele confers a selective advantage for tumor growth. The haploinsufficiency is further supported by that Bcl11b(+/-) mouse embryos were impaired in thymocyte development and survival. These results indicate relevance of BCL11B aberration to human leukemogenesis.


Subject(s)
DNA-Binding Proteins/genetics , Repressor Proteins/genetics , Tumor Suppressor Proteins/genetics , Animals , Base Sequence , Cell Differentiation , DNA Primers , DNA-Binding Proteins/immunology , Flow Cytometry , Gamma Rays , Genes, Tumor Suppressor , Genetic Predisposition to Disease , Genotype , Haplotypes , Lymphoma/genetics , Mice , Mice, Inbred BALB C , Repressor Proteins/immunology , Thymus Gland/cytology , Thymus Neoplasms/genetics , Tumor Suppressor Proteins/immunology
14.
Biochem Biophys Res Commun ; 354(1): 209-15, 2007 Mar 02.
Article in English | MEDLINE | ID: mdl-17210131

ABSTRACT

Mouse strains exhibit different susceptibilities to gamma-ray-induced thymic lymphomas. Our previous study identified Mtf-1 (metal responsive transcription factor-1) as a candidate susceptibility gene, which is involved in the radiation-induced signaling pathway that regulates the cellular reactive oxygen species (ROS). To reveal the mechanism for the increased susceptibility conferred by Mtf-1 locus, we examined early effects of gamma-ray on ROS levels in vivo and its difference between Mtf-1 susceptible and resistant congenic mice. Here, we show the detection of clonally growing thymocytes at 4 weeks after irradiation, indicating the start of clonal expansion at a very early stage. We also show that large thymocytes with higher ROS levels and a proliferation capacity were more numerous in the Mtf-1 susceptible mice than the resistant mice when examined at 7 days after irradiation, although such tendency was not found in mice lacking one allele of Bcl11b tumor suppressor gene. This high retention of the large thymocytes, at a high risk for ROS-induced mutation, is a compensatory proliferation and regeneration response to depletion of the thymocytes after irradiation and the response is likely to augment the development of prelymphoma cells leading to thymic lymphomas.


Subject(s)
DNA-Binding Proteins/metabolism , Gamma Rays/adverse effects , Lymphoma/physiopathology , Radiation Injuries/physiopathology , Reactive Oxygen Species/metabolism , Thymus Gland/metabolism , Thymus Gland/radiation effects , Transcription Factors/metabolism , Animals , Dose-Response Relationship, Radiation , Genetic Predisposition to Disease/etiology , Genetic Predisposition to Disease/genetics , Lymphoma/etiology , Mice , Mice, Inbred BALB C , Radiation Dosage , Radiation Injuries/etiology , Radiation Tolerance , Transcription Factor MTF-1
15.
J Immunol ; 176(10): 5871-9, 2006 May 15.
Article in English | MEDLINE | ID: mdl-16670294

ABSTRACT

Bcl11b(-/-) mice show developmental arrest at the CD44(-)CD25(+) double-negative 3 (DN3) or immature CD8(+)single-positive stage of alphabeta T cell. We have performed detailed analysis of sorted subsets of Bcl11b(-/-) thymocytes, DN3 and CD44(-)CD25(-) double-negative 4 (DN4) cells. Surface expression of TCRbeta proteins was not detected in DN3 thymocytes and markedly reduced in DN4 thymocytes, whereas expression within the cell was detected in both, suggesting some impairment in processing of TCRbeta proteins from the cytoplasm to the cell surface. This lack of expression, resulting in the absence of pre-TCR signaling, could be responsible for the arrest, but the transgenic TCRbeta or TCRalphabeta expression on the cell surface failed to promote transition from the DN3 to CD4(+)CD8(+) double-positive stage of development. This suggests that the pre-TCR signal cannot compensate the deficiency of Bcl11b for development. Bcl11b(-/-) DN3 thymocytes showed normal DNA rearrangements between Dbeta and Jbeta segments but limited DNA rearrangements between Vbeta and DJbeta without effect of distal or proximal positions. Because this impairment may be due to chromatin accessibility, we have examined histone H3 acetylation in Bcl11b(-/-) DN3 cells using chromatin immunoprecipitation assay. No change was observed in acetylation at the Vbeta and Dbeta gene locus. Analysis of Bcl11b(-/-) DN4 thymocytes showed apoptosis, accompanied with lower expression of anti-apoptotic proteins, Bcl-x(L) and Bcl-2, than wild-type DN4 thymocytes. Interestingly, the transgenic TCRalphabeta in those cells reduced apoptosis and raised their protein expression without increased cellularity. These results suggest that Bcl11b deficiency affects many different signaling pathways leading to development arrests.


Subject(s)
Apoptosis/immunology , DNA-Binding Proteins/deficiency , Receptors, Antigen, T-Cell, alpha-beta/biosynthesis , Receptors, Antigen, T-Cell, alpha-beta/genetics , T-Lymphocytes/metabolism , Tumor Suppressor Proteins/deficiency , Animals , Apoptosis/genetics , Cell Differentiation/genetics , Cell Differentiation/immunology , Cells, Cultured , DNA-Binding Proteins/genetics , Mice , Mice, Inbred BALB C , Mice, Inbred C57BL , Mice, Knockout , Mice, Transgenic , Receptors, Antigen, T-Cell, alpha-beta/deficiency , Receptors, Antigen, T-Cell, alpha-beta/physiology , Repressor Proteins/genetics , T-Lymphocytes/cytology , T-Lymphocytes/immunology , Tumor Suppressor Proteins/genetics
16.
Biochem Biophys Res Commun ; 340(2): 517-25, 2006 Feb 10.
Article in English | MEDLINE | ID: mdl-16376299

ABSTRACT

The rat aldolase B promoter acts as a replication origin in vivo, as well as an autonomously replicating sequence (ARS). Here, we examined roles of a polypurine stretch (site PPu) in this origin, which is indispensable to the ARS activity. Purification of site PPu-binding protein revealed that site PPu binds Puralpha and Purbeta, i.e., single-stranded DNA-binding proteins whose roles in replication have been implicated, but less clear. Biochemical analyses showed that site PPu even in a longer DNA fragment is unstable in terms of double-helix, implying that Puralpha/beta may stabilize single-stranded state. Deletion of site PPu from the origin DNA, which was ectopically positioned in the mouse chromosome, significantly reduced replicator activity. Chromatin immunoprecipitation experiments showed that deletion of site PPu abolishes binding of the Puralpha/beta proteins to the origin. These observations suggest functional roles of site PPu and Puralpha/beta proteins in replication initiation.


Subject(s)
DNA Replication/physiology , DNA-Binding Proteins/metabolism , Fructose-Bisphosphate Aldolase/biosynthesis , Fructose-Bisphosphate Aldolase/genetics , Replication Origin/genetics , Transcription Factors/metabolism , Amino Acid Sequence , Animals , Binding Sites/genetics , Cell Line, Tumor , Fructose-Bisphosphate Aldolase/metabolism , L Cells , Mice , Molecular Sequence Data , Mutagenesis, Site-Directed , Promoter Regions, Genetic , Rats , Replication Origin/physiology
17.
Radiat Res ; 163(2): 159-64, 2005 Feb.
Article in English | MEDLINE | ID: mdl-15658891

ABSTRACT

Mouse thymic lymphomas are readily induced by radiation and also arise without irradiation when the mice are null in Trp53 functions. In the present study, spontaneous thymic lymphomas in Trp53-/- mice were compared to those arising in irradiated Trp53+/- mice, revealing three features characteristic of the spontaneous lymphomas. (1) Mp53D2, a Trp53 modifier that affects the latent period of radiogenic thymic lymphomas in Trp53+/- mice, had no effect on the development of spontaneous lymphomas. (2) A sex difference in the latency was found. (3) A marked difference was noted in the frequency of allelic loss at the Ikaros gene on chromosome 11, encoding a transcription factor required for normal lymphocyte development and differentiation; 2% in the lymphomas of Trp53-/- mice and 78% in the radiogenic lymphomas of Trp53+/- mice, suggesting that loss of Trp53 may reduce the requirement for the loss of Ikaros for lymphomagenesis. Furthermore, allelic loss analysis on chromosome 19 localized a region that may harbor an unknown tumor suppressor gene. These results suggest intricate steps of lymphomagenesis influenced by the presence or absence of Trp53.


Subject(s)
Genetic Predisposition to Disease/genetics , Lymphoma/metabolism , Neoplasms, Radiation-Induced/metabolism , Thymus Neoplasms/metabolism , Tumor Suppressor Protein p53/genetics , Tumor Suppressor Protein p53/metabolism , Animals , Female , Lymphoma/genetics , Male , Mice , Mice, Inbred C57BL , Neoplasms, Radiation-Induced/genetics , Radiation Dosage , Thymus Neoplasms/genetics , Whole-Body Irradiation/adverse effects
18.
Oncogene ; 24(3): 399-406, 2005 Jan 13.
Article in English | MEDLINE | ID: mdl-15516976

ABSTRACT

Genetic predisposition to cancers is significant to public health because a high proportion of cancers probably arise in a susceptible human subpopulation. Using a mouse model of gamma-ray-induced thymic lymphomas, we performed linkage analysis and haplotype mapping that suggested Mtf-1, metal-responsive transcription factor-1 (Mtf-1), as a candidate lymphoma susceptibility gene. Sequence analysis revealed a polymorphism of Mtf-1 that alters the corresponding amino acid at position 424 in the proline-rich domain from a serine in susceptibility strains to proline in resistant strains. The transcriptional activity of Mtf-1 encoding serine and proline was compared by transfecting the DNA to Mtf-1-null cells, and the change to proline conferred a higher metal responsiveness in transfections. Furthermore, the resistant congenic strains possessing the Mtf-1 allele of proline type exhibited higher radiation inducibility of target genes than susceptible background strains having the Mtf-1 allele of serine type. Since products of the targets such as metallothionein are able to suppress cellular stresses generated by irradiation, these results suggest that highly inducible strains having Mtf-1 of proline type are refractory to radiation effects and hence are resistant to lymphoma development.


Subject(s)
Lymphoma/genetics , Neoplasms, Radiation-Induced/genetics , Polymorphism, Single Nucleotide/genetics , Thymus Neoplasms/genetics , Transcription Factors/genetics , Amino Acid Sequence , Amino Acid Substitution , Animals , Base Sequence , Cell Line, Tumor , DNA Primers , DNA-Binding Proteins , Disease Models, Animal , Genetic Markers , Genetic Predisposition to Disease , Genetic Variation , Immunity, Innate , Mice , Mice, Inbred BALB C , Proline , Serine , Transfection , Transcription Factor MTF-1
19.
Carcinogenesis ; 25(6): 1069-75, 2004 Jun.
Article in English | MEDLINE | ID: mdl-14754877

ABSTRACT

Mouse thymic lymphomas induced by gamma-irradiation exhibited homozygous deletions of the Rit1/Bcl11b tumor suppressor gene on chromosome 12 at high frequencies. Internal deletions of one allele were frequently accompanied by loss of the other allele. In order to elucidate the mechanism of these internal deletions, the sites of breakage and rejoining were examined by PCR mapping and sequencing. The 5' site of the deletions clustered within an approximately 5 kb region of intron 1 and the 3' site was confined to a site in intron 3. These sites contained P and/or N nucleotides and cryptic sequences recognizable by the RAG1/2 recombinase in the vicinity. This suggests that the Rit1 intragenic deletions were generated by endogenous illegitimate V(D)J recombinase activity and such aberrant recombination was also detected by nested PCR of DNA from the thymus of unirradiated mice but not of RAG2-deficient mice. A rough estimate indicated that there reside as many as 10(3)-10(4) thymocytes having Rit1 deletions, assuming the presence of 10(8) thymocytes in the thymus of unirradiated mice. Moreover, the recombination frequency was not affected by gamma-irradiation. These results show no effect of radiation on Rit1 mutations and suggest an indirect mechanism for its role in lymphomagenesis.


Subject(s)
Gene Deletion , Lymphoma/genetics , Neoplasms, Radiation-Induced/genetics , Thymus Gland/metabolism , Thymus Neoplasms/genetics , VDJ Recombinases/metabolism , ras Proteins/genetics , Animals , Base Sequence , DNA Primers , Female , Gamma Rays , Lymphoma/enzymology , Lymphoma/etiology , Male , Mice , Neoplasms, Radiation-Induced/enzymology , Neoplasms, Radiation-Induced/etiology , Thymus Gland/enzymology , Thymus Neoplasms/enzymology , Thymus Neoplasms/etiology
20.
Biochem Biophys Res Commun ; 313(3): 489-95, 2004 Jan 16.
Article in English | MEDLINE | ID: mdl-14697215

ABSTRACT

Chromatin state of a 2-Mb region harboring Rit1/Bcl11b on mouse chromosome 12 was examined using two distinct methods. One is ChIP assay examining the degree of enrichment with histone H3 methylated at lysine 9 (H3-mLys9) in chromatin and the other is H/E (heterochromatin/euchromatin) assay that measures a chromatin condensation state by using centrifugation. The ChIP assay showed that a 50-kb interval covering the gene and an upstream region constituted chromatin enriched with unmethylated H3-mLys9 in cells expressing Rit1 compared to cells not expressing Rit1. In contrast, regions other than the 50-kb interval did not show much difference in the enrichment between the two different types of cells. On the other hand, H/E assay of two expressing and two non-expressing tissues provided compatible fractionation patterns, suggesting that the chromatin condensation state detected by H/E assay is correlated with the chromatin state controlled by histone H3 tail modification linked to gene expression. These results indicate that the centrifugation-based H/E assay should provide a new approach to the regulation of chromatin structure with respect to its condensation state, complementing ChIP assays.


Subject(s)
Chromatin/chemistry , Histones/chemistry , ras Proteins/genetics , Animals , Antibodies/chemistry , Blotting, Northern , Blotting, Western , Cell Division , Cell Line, Tumor , Centrifugation/methods , Chromatin/metabolism , DNA/chemistry , Liver/metabolism , Lysine/chemistry , Methylation , Mice , Models, Genetic , Polymerase Chain Reaction , Precipitin Tests , Thymus Gland/metabolism , Tissue Distribution
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