scholarly journals Protein expression, mRNA expression and gene amplification of DNA methyltransferase 1 in endometrial tumor tissues

2013 ◽  
Vol 1 (3) ◽  
pp. 423-429 ◽  
Author(s):  
SATOSHI IKEDA ◽  
JYOJI IMURA ◽  
KEIKO SUZUKI
Keyword(s):  
Protein Expression ◽  
Mrna Expression ◽  
Gene Amplification ◽  
Dna Methyltransferase ◽  
Dna Methyltransferase 1 ◽  
Tumor Tissues ◽  
Endometrial Tumor ◽  
Methyltransferase 1

99 EFFECT OF SUPPRESSION OF BOVINE DNA METHYLTRANSFERASE 1 ON EMBRYONIC DEVELOPMENT AND EXPRESSION OF IMPRINTED GENES

2009 ◽  
Vol 21 (1) ◽  
pp. 150
Author(s):  
P. Wilaiphan ◽  
F. Rings ◽  
M. Hoelker ◽  
E. Tholen ◽  
C. Phatsara ◽  
...  
Keyword(s):  
Embryonic Development ◽  
Protein Expression ◽  
Dna Methyltransferase ◽  
Critical Role ◽  
Apoptotic Index ◽  
Imprinted Genes ◽  
Dna Methyltransferase 1 ◽  
Relative Standard ◽  
Blastocyst Rate ◽  
Methyltransferase 1

DNA methyltransferase 1 (DNMT1) is believed to be involved in DNA methylation, which is a well-characterized epigenetic modulator shown to have essential functions in germ line and embryonic genome imprinting. This study was conducted to investigate the consequences of suppressing and inhibiting DNMT1 on the development, level of apoptosis, and expression of imprinted genes in pre-implantation bovine embryos. In vitro-produced zygotes were categorized into 4 groups; namely, those injected with Smartpool siRNA (SpsiRNA; Dharmacon Inc., Chicago, IL) (n = 800), 5aza-2′-deoxycytidine (5-AZA; Sigma, St. Louis, MO) (n = 864), nuclease-free water (n = 850), and uninjected control (n = 755). The mRNA expression data were generated using RT-PCR based on the relative standard curve method employing glyceraldehyde-3-phosphate dehydrogenase (GAPDH) as a normalizer. Apoptotic index (API) was calculated by dividing the number of apoptotic cells to total cell number. The proportion of 2-, 4-, 8-cell and 2-, 4-, 8-, and 16-cell embryos was assessed 48 and 72 h, respectively, post-micro injection (pmi), whereas blastocyst rate was assessed at Day 8 pmi. Data on embryonic development and the relative mRNA abundance were analyzed using ANOVA followed by a multiple pair-wise mean comparison using Tukey test. The proportion of 2-, 4-, and 8-cell embryos at 48 h pmi was not significant among treatment groups. However, the proportion of the 8-cell embryos was significantly lower (P < 0.05) in SpsiRNA (16.3 ± 4.5) and 5-AZA injected groups (17.7 ± 4.9) compared with water-injected (26.8 ± 2.9) and uninjected controls (30.7 ± 6.2). The lowest total blastocyst rate (P < 0.05) was observed in the 5-AZA treatment group (16.9 ± 4.9) compared with SpsiRNA (23.4 ± 2.1) and water-injected (24.1 ± 5.3) and uninjected controls (29.4 ± 2.1). Microinjection of SpsiRNA reduced the target mRNA by 80 and 50% in 8-cell and blastocyst stage embryos, respectively, compared with uninjected control, and the protein expression level was also reduced at 8-cell embryos as confirmed by Western blotting. Injection of 5-AZA had no significant effect on mRNA or protein expression. The greatest API (P < 0.05) was found in SpsiRNA (4.2 ± 2.0) and 5-AZA (4.1 ± 1.7) injected groups compared with water-injected (2.8 ± 2.1) and uninjected controls (2.9 ± 2.3). The relative expression study also showed that microinjection of SpsiRNA and 5-AZA increased the expression of IGF2 (by 67 and 55%), IGF2R (29 and 30%), and IGFPB-4 (22 and 24%), respectively, compared with uninjected control, without affecting the expression of both IGF2R and IGFPB-4. In conclusion, suppression of DNMT1 resulted in lower proportion of 8-cell embryos, reduced blastocyst rate, and increased apoptotic index, and affected the expression of some imprinted genes, demonstrating a critical role of this gene in bovine embryonic development.

scholarly journals RX-3117 (Fluorocyclopentenyl-Cytosine)-Mediated Down-Regulation of DNA Methyltransferase 1 Leads to Protein Expression of Tumor-Suppressor Genes and Increased Functionality of the Proton-Coupled Folate Carrier

2020 ◽  
Vol 21 (8) ◽  
pp. 2717 ◽  
Author(s):  
Dzjemma Sarkisjan ◽  
Joris R. Julsing ◽  
Btissame El Hassouni ◽  
Richard J. Honeywell ◽  
Ietje Kathmann ◽  
...  
Keyword(s):  
Tumor Suppressor ◽  
Protein Expression ◽  
Tumor Suppressor Genes ◽  
Dna Methyltransferase ◽  
Functional Assay ◽  
Dna Methyltransferase 1 ◽  
Suppressor Genes ◽  
Resistant Variant ◽  
E Cadherin ◽  
Methyltransferase 1

(1) Background: RX-3117 (fluorocyclopentenyl-cytosine) is a cytidine analog that inhibits DNA methyltransferase 1 (DNMT1). We investigated the mechanism and potential of RX-3117 as a demethylating agent in several in vitro models. (2) Methods: we used western blotting to measure expression of several proteins known to be down-regulated by DNA methylation: O6-methylguanine-DNA methyltransferase (MGMT) and the tumor-suppressor genes, p16 and E-cadherin. Transport of methotrexate (MTX) mediated by the proton-coupled folate transporter (PCFT) was used as a functional assay. (3) Results: RX-3117 treatment decreased total DNA-cytosine-methylation in A549 non-small cell lung cancer (NSCLC) cells, and induced protein expression of MGMT, p16 and E-cadherin in A549 and SW1573 NSCLC cells. Leukemic CCRF-CEM cells and the MTX-resistant variant (CEM/MTX, with a deficient reduced folate carrier) have a very low expression of PCFT due to promoter hypermethylation. In CEM/MTX cells, pre-treatment with RX-3117 increased PCFT-mediated MTX uptake 8-fold, and in CEM cells 4-fold. With the reference hypomethylating agent 5-aza-2′-deoxycytidine similar values were obtained. RX-3117 also increased PCFT gene expression and PCFT protein. (4) Conclusion: RX-3117 down-regulates DNMT1, leading to hypomethylation of DNA. From the increased protein expression of tumor-suppressor genes and functional activation of PCFT, we concluded that RX-3117 might have induced hypomethylation of the promotor.

scholarly journals DNA methyltransferase 1 regulates reelin mRNA expression in mouse primary cortical cultures

2005 ◽  
Vol 102 (5) ◽  
pp. 1749-1754 ◽  
Author(s):  
J. S. Noh ◽  
R. P. Sharma ◽  
M. Veldic ◽  
A. A. Salvacion ◽  
X. Jia ◽  
...  
Keyword(s):  
Mrna Expression ◽  
Dna Methyltransferase ◽  
Dna Methyltransferase 1 ◽  
Cortical Cultures ◽  
Methyltransferase 1

scholarly journals Increased DNA methyltransferase 1 (DNMT1) protein expression in precancerous conditions and ductal carcinomas of the pancreas

2005 ◽  
Vol 96 (7) ◽  
pp. 403-408 ◽  
Author(s):  
Dun-Fa Peng ◽  
Yae Kanai ◽  
Morio Sawada ◽  
Saori Ushijima ◽  
Nobuyoshi Hiraoka ◽  
...  
Keyword(s):  
Protein Expression ◽  
Dna Methyltransferase ◽  
Precancerous Conditions ◽  
Dna Methyltransferase 1 ◽  
Ductal Carcinomas ◽  
Methyltransferase 1
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